Objective To discuss the effect of long non-coding RNA （lncRNA） gastric cancer related transcript 2（GACAT2） gene on the proliferation and stemness of the lung cancer A549 and H1299 cells， and to clarify the effect of lncRNA GACAT2 gene on the occurrence and development of lung cancer. Methods The full-length sequence of lncRNA GACAT2 gene was amplified by real-time fluorescence quantitative PCR （RT-qPCR） method， and was cloned into the pc3.1 （+） eukaryotic expression vector. The construction of GACAT2 over-expression vector was confirmed by bacterial liquid PCR method and gene sequencing. The pc3.1-GACAT2 over-expression plasmid and control plasmid pc3.1 were separately transfected into the human lung cancer A549 and H1299 cells. The A549 and H1299 cells with stable over-expression of GACAT2 were established by G418 screening， and the cells were divided into blank vector group （transfected with pc3.1 empty vector） and pc3.1-GACAT2 group （transfected with pc3.1-GACAT2 recombinant plasmid）. RT-qPCR method was used to detect the expression levels of GACAT2 mRNA in the A549 and H1299 cells in two groups；CCK-8 assay was used to detect the proliferation activities of the cells in two groups； cell clone formation assay was used to detect the clone formation numbers of the cells in two groups； stem cell sphere formation experiment was used to detect the sphere formation numbers of the A549 and H1299 cells in two groups. Results The eukaryotic over-expression vector of GACAT2 was successfully constructed. Compared with blank vector group， the expression levels of GACAT2 mRNA in the A549 and H1299 cells in pc3.1-GACAT2 group were increased （P<0.05 or P<0.01）， the cell proliferation activities were decreased （P<0.05 or P<0.01），the clone formation numbers were decreased （P<0.05 or P<0.01）， and the sphere formation numbers were decreased （P<0.05 or P<0.01）. Conclusion Over-expression of human lncRNA GACAT2 gene can inhibit the proliferation and stemness of the lung cancer A549 and H1299 cells.

Objective To discuss the change of sensitivity of the non-small-cell lung cancer （NSCLC） A549 cells to doxorubicin （Dox） after silencing forkhead protein 3 （FOXP3） gene，and to clarify its mechanism involved in Dox resistance. Methods The human NSCLC A549 cells were transfected with FOXP3 small interfering RNA（siRNA ）by lipofectamine method. The cells were divided into blank control group （without transfection）， si-NC group （transfected with control-siRNA）， and si-FOXP3 group （transfected with FOXP3-siRNA）. Western blotting and immunofluorescence methods were used to detect the expression levels of FOXP3 protein in the A549 cells in various groups；the proliferation activities and half-maximal inhibitory concentration （IC₅₀） values of the A549 cells in various groups were detected by CCK-8 method. The A549 cells were treated with 0， 10， and 20 μmol·L^－1 DAPT， and regarded as 0， 10， and 20 μmol·L^－1 DAPT groups， respectively. Additionally， the A549 cells were treated with 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT， and regarded as 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups， respectively. Western blotting method was used to detect the expression levels of Notch1，Hes1， and FOXP3 proteins in the A549 cells in various groups； the IC₅₀ values and expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 0， 10， and 20 μmol·L^－1 DAPT groups were detected by CCK-8 and Western blotting methods；the expression levels of FOXP3， P-glycoprotein（P-gp）， Notch1， and Hes1 proteins in the A549 cells in 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups were detected by Western blotting method. Results Compared with blank control and si-NC groups， the expression level of FOXP3 protein in the A549 cells in si-FOXP3 group was significantly decreased （P<0.01）， the proliferative activity and IC₅₀ value were decreased （P<0.05 or P<0.01），and the expression levels of Notch1，Hes1 and FOXP3 proteins were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 10 and 20 μmol·L^－1 DAPT groups were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox group were significantly increased （P<0.01）. Compared with 1.0 mg·L^－1 Dox group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT group were significantly decreased （P<0.01）. Conclusion Silencing FOXP3 can enhance the sensitivity of the NSCLC cells to Dox， and its mechanism is related to the inhibition of the Notch1/Hes1 signaling pathway.

Objective To discuss the therapeutic effect of Saposhnikoviae Radix on rheumatoid arthritis and its effect on the nuclear factor-κB （NF-κB） signaling pathway，and to provide the basis for the treatment of rheumatoid arthritis. Methods Fifty rats were randomly divided into control group， model group， dexamethasone group， low dose of Saposhnikoviae Radix group， and high dose of Saposhnikoviae Radix group，and there were 10 rats in each group； except for control group， the rats in the other groups were given intradermal injection of type Ⅱ collagen at the root of the tail and left posterior foot to prepare the rheumatoid arthritis models； after the models were prepared successfully， the rats in dexamethasone group were given dexamethasone （2 mg·kg^－1），the rats in low and high doses of Saposhnikoviae Radix groups were given Saposhnikoviae Radix wild product （0.45 and 0.90 g·kg^－1）， and equal volume of normal saline was given to the rats in control group and model group.The arthritis index（AI） and foot swelling degrees of the rats in various groups were recorded， the organ coefficients of the rats in various groups were calculated； enzyme-linked immunosorben assay （ELISA） method was used to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in serum of the rats in various groups， and the expression levels of NF-κB，NF-κB inhibitor α （IkB-α）， IκB-α kinase β （IKK-β）， cyclooxidase-2 （COX-2），and TNF-α proteins in synovial tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group， the AI score and swelling degree of foot of the rats in model group were significantly decreased （P<0.01）， while the thymus index was significantly increased （P<0.01）； compared with model group， the AI score and swelling degree of foot of the rats in dexamethasone group and low and high doses of Saposhnikoviae Radix groups were significantly decreased （P<0.05 or P<0.01）， and the thymus indexes were significantly decreased（P<0.01）.The ELISA results showed that compared with control group，the levels of TNF-α， IL-1β，and IL-6 in serum of the rats in model group were increased（P<0.01）； compared with model group，the levels of TNF-α， IL-1β，and IL-6 in serum of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were significantly decreased （P<0.01）.The HE staing results showed that the synovial tissue of the rats in control group was normal，and the synovial tissue of the rats in model group showed hyperplasia accompanied with extensive infiltration of inflammatory cells； compared with model group， the degrees of proliferation of synovial tissue and infiltration of inflammatory cells of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were decreased.The Western blotting results showed that compared with control group， the expression levels of NF-κB，IKK-β，COX-2， and TNF-α proteins in synovial tissue of the rats in model group were increased （P<0.01），the expression level of IκB-α protein was decreased （P<0.01）；compared with model group， the expression levels of NF-κB，IKK-β，COX-2， and TNF-α proteins in synovial tissue of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were decreased （P<0.01），the expression level of IκB-α protein was increased （P<0.01）. Conclusion Saposhnikoviae Radix wild product can improve the joint injury by reducing the levels of proinflammatory cytokines and inhibiting the excessive activation of NF-κB signaling pathway in synovial tissue， thus playing a therapeutic role in the rheumatoid arthritis.

Objective To screen the differentially expressed genes in type 2 diabetes mellitus model rats by transcriptomics sequencing， and to preliminarily explore the relationship between the differentially expressed genes and the syndrome manifestations of deficiency of both Qi and Yin. Methods Eighteen male SD rats were randomly selected as control group，and the other rats were fed with high-fat and high-sugar diet and injected with streptozotocin （STZ） in the abdomen to establish the type 2 diabetes models. On the 6th week， 31 successfully modeling rats were divided into model group （n=14） and drug evidence group （n=17） according to the principle of blood glucose and body mass balance. The rats in model group were fed with high sugar and high fat diet sequentially.The rats in drug evidence group were given Xiaokeling 2.2 g·kg^－1·d^－1 by gavage；the experiment last for 21 weeks.The body masses， food intakes， water intakes， mental state scores， sport scores， foot temperatures， grasping force， pulse amplitudes， respiratory rates， and tongue image integral of the rats were detected and the physiological signs of the rats during model preparation were evaluated.The fasting blood glucose（FBG） levels of the rats in various groups were detected by blood glucose meter. After the last administration， the rats were anesthetized， the blood samples were collected and the serum samples were separated.The levels of triglyceride （TG） and total cholesterol （TC） of the rats were detected by oxidase method.The levels of high density lipoprotein cholesterol （HDL-c） and low density lipoprotein cholesterol（LDL-c）of the rats were detected by direct method，and the levels of CD4， CD8，cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） of the rats were detected by enzyme-linked immunosorbent assay （ELISA） method.The liver tissue of the rats was obtained，and the differentially expressed genes between model group and control group and between model group and drug evidence group were screened by transcriptomic sequencing. Results Compared with blank group， the physiological signs of the rats in model group were basically consistent with the syndrome manifestations of Qi and Yin deficiency； compared with model group， the some physiological signs of Qi and Yin deficiency of the rats in drug evidence group were significantly improved. Compared with blank group， the FBG，serum TC， TG and LDL-c levels of the rats in model group were significantly increased （P<0.01）， and the serum HDL-c level had no significant difference（P>0.05）；compared with model group， there were no significant differences in the above indexes of the rats in drug evidence group （P>0.05）.Compared with blank group， the level of serum CD4 and the CD4/CD8 ratio of the rats in model group were significantly decreased （P<0.01）， while the level of serum CD8 had no significant difference（P>0.05）；the cAMP level in serum and the ratio of cAMP/cGMP were obviously increased （P<0.01）， while the cGMP level in serum was significantly decreased （P<0.01）；compared with model group， the level of serum CD4 and the CD4/CD8 ^{ratio of the rats in drug evidence group were remarkably increased （P<0.01）， the cGMP level in serum was increased（P<0.01） and the cAMP/cGMP ratio was significantly decreased（P<0.01）， the cAMP level in serum had no significant difference （P>0.05）. Compared with blank group， the up-regulated expressed genes of the rats in model group mainly included early growth response factor 2 （Egr2），insulin-like growth factor binding protein 1 （Igfbp1） and a disintegrin and metallopeptidase with thrombospondin motifs 4 （Adamts4）； the down-regulated expressed genes mainly included G₀/G₁ switch 2 （G0s2）， midline 1 interaction protein 1 （Mid1ip1）， and basic helix-loop-helix family e40 （Bhlhe40）；compared with model group， the levels of the above differentially expressed genes in drug evidence group showed the opposite trend. Conclusion The syndrome of type 2 diabetes mellitus combined with deficiency of Qi and Yin is associated with up-regulation of Egr2， Igfbp1 and Adamts4 and down-regulation of G0s2， Mid1ip1，and Bhlhe40.}

Objective To investigate the curative effect of osimertinib combined with savolitinib targeted therapy in the patients with non-small cell lung cancer（NSCLC） and central nervous system（CNS） metastasis with acquired resistance to epidermal growth factor receptor（EGFR）-tyrosine kinase inhibitor（TKI）， and to provide the reference for the targeted therapy for the disease. Methods The clinical data of one NSCLC complicated with CNS metastasis patient with non-frameshift deletion mutation in exon 19 of the EGFR gene complicated with MET amplification were collected， and the clinical characteristics， treatment methods， drug resistance mechanisms of targeted therapy， and treatment methods after drug resistance were analyzed combined with the literature review. Results The patient， female，47 years old， was diagnosed as lung adenocarcinoma in our hospital 3 years ago. The genetic test results showed that there was a non-frameshift deletion mutation in exon 19 of the EGFR gene， and the patient was given icotinib targeted therapy. After 18 months of treatment， the genetic test results showed the patient had the Exon20-T790M mutation， and the patient received oral osimertinib targeted therapy. Twelve months later， the disease progressed and CNS metastasis was found by the re-examination， and the genetic test showed the patient had the mesenchymal-epidermal transforming factor（MET） amplification，the patient received oral osimertinib combined with anlotinib targeted therapy. More than 2 months later， the re-examination results found that the disease had further progress， and the treatment plan was adjusted into osimertinib combined with savolitinib targeted therapy. After 1 month and 4 months， the re-examination results of chest CT and head MRI showed that the lung lesions and brain lesions were significantly smaller than before. Conclusion For the NSCLC brain metastases patients with EGFR-TKI resistance complicated with MET amplification after long-term targeted therapy， the combination of osimertinib and savolitinib targeted therapy can significantly control the progress of the lung and CNS disease in the short term.

Objective To discuss the protective effect of Modified Xiao-Xian-Xiong Decoction （MXD） on liver injury in the rats with type 2 diabetes mellitus（T2DM）， and to clarify its possible mechanism. Methods Fifty rats were fed with high sugar and high fat combined with intraperitoneal injection of streptozotocin （STZ） to establish the T2DM rat models.A total of 40 rats successful modeling were randomly divided into model group（given no drugs），metformin group（given 200 mg·kg^－1 metformin）， low dose of MXD group （given 630 mg·kg^－1 MXD），and high dose of MXD group（given 1 260 mg·kg^－1 MXD）， and there were 10 rats in each group； another 10 rats were selected as control group（given standard feed）. The body masses of the rats in various groups were detected；the fasting blood glucose （FBG） levels of the rats in various groups were detected； the activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum of the rats in various groups were detected by fully automated biochemical analyzer；the pathomorphology of liver tissue of the rats in various groups were detected by HE staining； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin 1β （IL-1β）， interleukin 6 （IL-6）， tumor necrosis factor α （TNF-α） in serum； the levels of superoxide dismutase （SOD）， reductive glutathione （GSH）， and malondialdehyde （MDA） in liver tissue of the rats in various groups were measured by spectrophotometer；the expression levels of nuclear factor E2 related factor 2 （Nrf2） and heme oxygenase （HO-1） proteins in liver tissue of the rats in various groups were detected by Western blotting method. Results The rats in control group had moderate feeding and water intake，and the above indexes of the rats had no sudden increasing and decreasing；the rats in model group had typical symptoms of diabetes；compared with model group，the diabetes symptoms of the rats in meftormin group，low dose of MXD group，and high dose of MXD group had been improved to various degrees.Compared with control group， the body masses of the rats in model group were significantly decreased at different time points（P<0.01）， while the FBG levels were increased （P<0.01）； compared wtih model group，the levels of FBG of the rats in meftormin group，low dose of MXD group，and high dose of MXD group were decreased in the 2nd，3rd and 4th weeks after administration，and the levels of FBG of the rats in low dose of MXD group were decreased in the 3rd and 4th weeks after administration（P<0.05 or P<0.01）.Compared with control group，the activities of AST and ALT in serum of the rats in model group were increased（P<0.01）；compared with model group，the activities of AST and ALT in serum of the rats in metformin group，low dose of MXD group，and high dose of MXD group were decreased（P<0.01）.The HE staining results showed that the morphology of liver cells of the rats in control group was normal，and there was no inflammatory cell infiltration；there was serious inflammatory cell infiltration in model group；there was a bit of inflammatory cell infiltration in meftormin group；there were almost no inflammatory cell infiltrations in low and high doses of MXD groups.The ELISA method results showed that the levels of serum IL-1β， IL-6， and TNF- α in serum of the rats in model group were significantly increased （P<0.01）.Compared with model group， the levels of serum IL-1β， IL-6， and TNF-α of the rats in metformin group， low dose of MXD and high dose of MXD groups were decreased（P<0.01）， while the activity of SOD， the level of GSH， and the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in model group were significantly decreased （P<0.01）.The spectrophotometer results showed that compared with control group， the activity of SOD， the level of GSH in liver tissue of the rats in model group were decreased（P<0.01）， and the level of MDA was significantly increased （P<0.01）；compared with model group，the activities of SOD and the levels of GSH in liver tissue of the rats in metformin group，low dose of MXD group，and high dose of MXD group were decreased（P<0.01），and the levels of MDA in liver tissue of the rats in low and high doses of MXD groups were decreased（P<0.05 or P<0.01）.The Western blotting results showed that compared with control group，the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in model group were decreased（P<0.01）；compared with model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in metformin group，low dose of MXD group，and high dose of MXD group were increased（P<0.05 or P<0.01）. Conclusion MXD has protective effect on liver injury of the rats with type 2 diabetes， and its mechanism may be related to anti-inflammation and activation of Nrf2/HO-1 signaling pathway，thereby inhibition of oxidative stress.

Objective To discuss the effect of miR-93-5p on the proliferation， migration， and invasion of the rheumatoid arthritis fibroblasts-like synoviocytes（RA-FLSs），and to elucidate its possible mechanism. Methods The rheumatoid arthritis（RA） patients （RA group，n=37） and joint trauma patients who underwent joint replacement surgery （control group，n=30） were selected as the subjects.The RA-FLSs were isolated from synovial tissue of the RA patients， and identified by immunofluorescence and flow cytometry. The RA-FLSs were divided into blank group， mimics NC group （transfected with miR-93-5p mimics NC）， mimics group （transfected with miR-93-5p mimics）， OE-NC group （transfected with ROCK2 over-expression empty plasmid）， OE-Rho related spiral coil protein kinase （ROCK）2 group （transfected with ROCK2 over-expression plasmid）， mimics+OE-NC group （co-transfected with miR-93-5p mimics and ROCK2 over-expression empty plasmids），and mimics+OE-ROCK2 group （co-transfected with miR-93-5p mimics and ROCK2 over-expression plasmids）. Real time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-93-5p and ROCK2 mRNA in synovial tissue of the patients in two groups and cells in various groups；CCK-8 method was used to detect the proliferation activities of the cells in various groups； EdU staining was used to detect the EdU positive rates of the cells in various groups；Transwell champer assay was used to detect the migration and invasion numbers of RA-FLSs in various groups； Western blotting method was used to detect the expression levels of ROCK2， Ki-67， proliferating cell nuclear antigen （PCNA），matrix metalloproteinase 2 （MMP-2）， and matrix metalloproteinase 9 （MMP-9） proteins in the cells in various groups； the targeting relationship between miR-93-5p and ROCK2 was verified by double Luciferase reporter gene experiment. Results The immunofluorescence assay results showed that the expression of Vimentin protein was positive.The flow cytometry detection results showed that the expression of CD55 on surface of the third generation RA-FLSs was positive， while the expressions of CD14 and CD68 were negative， confirming that the isolated cells were the RA-FLSs. The RT-qPCR results showed that compared with control group， the expression level of miR-93-5p in synovial tissue of the patients in RA group was significantly decreased（P<0.01）， while the expression level of ROCK2 mRNA was significantly increased （P<0.01）. Compared with blank group and mimics NC group， the expression level of miR-93-5p of the cells in mimics group was significantly increased （P<0.01）， while the expression levels of ROCK2 mRNA and protein were significantly decreased （P<0.01）. The CCK-8 method and EdU staining results showed that compared with mimics NC group， the proliferation activitiy and EdU positive rate of the cells in mimics group were significantly decreased （P<0.01）， while the proliferation activitiy and EdU positive rate of the cells in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the proliferation activitiy and EdU positive rate of the cells in mimics+OE-ROCK2 group were increased （P<0.01）. The Transwell champer assay results showed that compared with mimics NC group， the migration and invasion numbers of RA-FLSs in mimics group were significantly decreased （P<0.01）， while the migration and invasion numbers of RA-FLSs in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the migration and invasion numbers of RA-FLSs in mimics+OE-ROCK2 group were increased （P<0.01）. The Western blotting method results showed that compared with mimics NC group， the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in mimics group were significantly decreased （P<0.01）， while the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in mimics+OE-ROCK2 group were significantly increased （P<0.01）.There was a targeted binding site between miR-93-5p and ROCK2-3'-UTR. The double luciferase report experiment results showed that transfection of miR-93-5p mimic could significantly decrease the luciferase activity of the cells in ROCK2 wild type（ROCK2-WT） group（P<0.01）. Conclusion Over-expression of miR-93-5p inhibits the proliferation， migration， and invasion of the RA-FLSs targeted by down-regulation of ROCK2 expression.

Objective To discuss the effect of hypoxia inducible factor-1α （HIF-1α）/ reactive oxygen species （ROS） on the apoptosis and invasion of the non-small cell lung cancer A549 cells，and to clarify the related mechanism. Methods The A549 cells were cultured under hypoxia condition for 12， 24， 48 and 72 h to construct the hypoxia cell model. The expression levels of HIF-1α mRNA in the cells at different treatment time were detected by real-time fluorescence quantitative PCR（RT-qPCR） method， and the proliferation abilities of the cells were detected by CCK-8 assay；the hypoxia cell model was verified， and the optimal induction time was determined.The A549 cells were divided into control group（21%O₂）， model group（hypoxia induction）， NAC group（20 mmol·L^－1 NAC）， NAC+YC-1 group（20 mmol·L^－1 NAC+100 μmol·L^－1 YC-1） and YC-1 （100 μmol·L^－1 YC-1）group. The expression levels of HIF-1α and FPR1 mRNA and proteins were detected by RT-qPCR and Western blotting methods； the ROS levels were detected by flow cytometry； the autophagosome structure in the cells was observed by transmission electron microscope； the expressions of microtubule-associated protein light china 3 Ⅱ（LC3-Ⅱ） in the cells in various groups were detected by immunofluorescence； the apoptotic rates of the cells in various groups were detected by flow cytometry； and the number of invasion cells was detected by Transwell chamber assay. Results With the prolongation of hypoxia induction time， the HIF-1α mRNA expression levels in the A549 cells and cell proliferation abilities were increased（P<0.05 or P<0.01）， and the optimal hypoxia induction time was 24 h. The results of RT-qPCR and Western blotting methods showed that compared with control group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in model group were significantly increased（P<0.01）； compared with model group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased（P<0.01）.Compared with control group，the level of ROS in the cells in model group was significantly increased （P<0.01）； compared with model group，the ROS levels in the cells in NAC group， NAC+YC-1 group， and YC-1 group were significantly decreased（P<0.01）；compared with NAC group，the ROS level in the cells in NAC+YC-1 group was significantly decreased（P<0.01）.The transmission electron microscope results showed that the organelle structure of the cells in control group was intact and no autophagosome structure was seen； a large number of autophagosomes were seen in the cells in model group， obvious vacuoles were visible， and intracellular organelles were degraded； compared with model group， the number of autophagosomes in the cells in NAC group， NAC+YC-1 group and YC-1 group were decreased，among them the autophagosomes in the cells in NAC+YC-1 group were the least. The immunofluorescence assay results showed that the expression intensity of LC3-Ⅱ in the cells in model group was significantly increased compared with control group， and the expression intensities of LC3-Ⅱ in the cells in NAC group， NAC+YC-1 and YC-1 group were signficantly decreased compared with model group.Compared with control group， the apoptotic rate of the cells in model group was significantly decreased（P<0.01）； compared with model group， the apoptotic rates of the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly increased（P<0.01）； compared with NAC group，the apoptotic rate of the cells in NAC+YC-1 group was increased（P<0.01）.The results of Transwell chamber assay showed that compared with control group the number of the invasion cells in model group was significantly increased（P<0.01）； compared with model group， the numbers of invasion cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased （P<0.01）；compared with NAC group，the number of the in vasion cells in NAC+YC-1 group was decreased（P<0.01）. Conclusion Inhibition of HIF-1α/ROS under hypoxia condition can promote the apoptosis and inhibit cell invasion of cancer cells，and its mechanism may be realted to inhibiting the autophagy of lung cancer cells.

Objective To discuss the expression levels of long non-coding RNA paired box 8 antisense RNA 1 （PAX8-AS1） in the human colorectal cancer （CRC） tissue and its effects on the proliferation， apoptosis and invasion of the CRC cells，and to charify its mechanism. Methods The expression levels of PAX8-AS1 mRNA and miR-22-3p in cancer tissue and cancer cells of the CRC patients were detected by real-time fluorescence quantitative PCR （RT-qPCR） method.The PAX8-AS10 siRNA small molecule sequences and miR-22-3p inhibitors were transfected alone or co-transfected into the CRC cells， and the cells after transfection were divided into si-NC group （transfected with negative sequences）， si-PAX8-AS1 group （transfected with PAX8-AS1 siRNA）， si-PAX8-AS1+inhibitors NC group （co-transfected with PAX8-AS1 siRNA and inhibitors NC）， and si-PAX8-AS1+ inhibitors group （co-transfected with PAX8-AS1 siRNA and miR-22-3p inhibitors）， and the SW480 cells without any transfection were regarded as control group. The expression levels of PAX8-AS1 mRNA and miR-22-3p in the cells in various groups after transfection were measured by RT-qPCR method； the proliferation activities of the cells in various groups were detected by MTT method； the migration and invasion rates of the cells in various groups were measured by Transwell chamber assay； the apoptotic rates of cells in various groups were measured by flow cytometry； the expression levels of B-cell lymphoma-2（Bcl-2）， Bcl-2 associated X protein（Bax）， and cleaved Caspase-3 proteins in the cells in various groups were measured by Western blotting method； the target relationship between PAX8-AS1 and miR-22-3p was verified by dual luciferase reporter gene assay. Results The RT-qPCR results showed that compared with adjacent tissue，the expression level of PAX8-AS1 mRNA in cancer tissue was significantly increased（P<0.01）， and the expression level of miR-22-3p was significantly decreased（P<0.01）； compared with the human normal colonic epithelial cells NCM460，the PAX8-AS1 mRNA expression levels in the SW480， SW620， HT-29 and LoVo cells were all significantly increased （P<0.01）， while the miR-22-3p expression levels were all significantly decreased （P<0.05 or P<0.01）. Compared with control group and si-NC group， the expression level of PAX8-AS1 mRNA in the cells in si-PAX8-AS1 group was decreased（P<0.01）， and the expression level of miR-22-3p was significantly increased（P<0.01）； compared with si-NC group，the expression level of miR-22-3p in the cells in si-PAX8-AS1+inhibitors group was increased（P<0.01）.The MTT assay results showed that compared with control group，the proliferation activities of the cells in si-PAX8-AS1 group，si-PAX8-AS1+inhibitors NC group，and si-PAX8-AS1+inhibitors group were significantly decreased after culture for 48 and 72 h （P<0.01）； compared with si-NC group， the proliferation activities of the cells in si-PAX8-AS1+ inhibitors NC group were significantly decreased after culture for 48 and 72 h （P<0.01）.The flow cytometry results showed that compared with control group， the apoptotic rate of cells in si-PAX8-AS1 group was significantly increased （P<0.01）；compared with si-NC group，the apoptotic rate of cells in si-PAX8-AS1+ inhibitors NC group was significantly increased （P<0.01）.The Transwell chamber assay results showed that compared with control group， the rates of migration and invasion cells in si-PAX8-AS1 group were significantly decreased（P<0.01）； compared with si-NC group，the rates of migration and invasion cells in si-PAX8-AS1 group and si-PAX8-AS1+ inhibitors NC group were significantly decreased（P<0.01）.The Western blotting results showed that compared with control group and NC group， the expression levels of Bax and cleaved Caspase-3 proteins in the cells in si-PAX8-AS1 group were significantly increased（P<0.01）， and expression level of the Bcl-2 protein was significantly decreased（P<0.05）.The TargetScan predicted that there were target binding sites between PAX8-AS1 and miR22-3p.The dual luciferase reporter gene assay results showed that compared with the cells co-transfected with mimics NC， the luciferase activity in the PAX8-AS1-WT cells was significantly decreased after co-transfected with miR-22-3p mimics （P<0.01）. Conclusion The PAX8-AS1 is highly expressed in the human CRC tissue， and silencing the PAX8-AS1 expression can inhibit the proliferation， migration and invasion of the CRC cells and induce the apoptosis，and its mechanism may be ralated to up-regulation the miR-22-3p expression.

Objective To analyze the effect of rheumatoid arthritis （RA） on the phenotypes of plasma cells of gingiva and expression of receptor activator of nuclear factor-κB ligand（RANKL） in the patients with periodontitis，and to clarify its possible mechanism. Methods Sixty subjects who came to our hospital were selected and divided into healthy group， periodontitis group and periodontitis+RA group according to inclusion criteria， and there were 20 subjects in each group.The gingival index （GI）， probing depth （PD）， bleeding on probing （BOP） and clinical attachment loss （CAL） of the subjects were recorded. The expression levels of proliferation inducing ligand（APRIL） and B lymphocyte stimulator（BLyS） in gingiva tissue of the subjects in various groups were detected by real-time fluoresence quantatitive PCR（RT-qPCR） method；the percentages of CD38+ cells， CD138+ cells in gingiva tissue of the subjects in various groups were analyzed by flow cytometry；the percentages of RANKL+ cells in the gingival plasma cells and the RANKL levels in plasma cells of gingiva of the subjects in various groups were detected by flow cytometry and enzyme-linked immunosorbent assay（ELISA） methods； the growth of osteoclasts in the subjects in various groups was observed by anti-tartrate acid phosphatase （TRAP） staining； the expression levels of RANKL， receptor activator of nuclear factor-κB（RANK） and tumor necrosis factor associated factor 6（TRAF6） mRNA in plasma cells of gingiva of the subjects in various groups were detected by RT-qPCR method. Results Compared with healthy group， GI， PD， BOP and CAL of the patients in periodontitis group and periodontitis+RA group were significantly increased （P<0.05）； compared with periodontitis group，the PD and CAL of the patients in periodontitis+RA group were increased（P<0.05）.Compared with healthy group， the expression levels of APRIL and BLyS mRNA in gingiva tissue of the patients in periodontitis group and periodontitic+PA group were increased（P<0.05）；compared with periodontitis group，the expression level of APRIL mRNA of the patients in periodontitis+RA group was increased（P<0.05）.Compared with healthy group， the percentages of CD38+ cells and CD138+ cells in gingiva tissue of the patients in periodontitis group and periodontitis+RA group were significantly increased（P<0.05）；compared with periodontitis group， the percentage of CD138+ cells in gingiva tissue of the patients in periodontitis+RA group was increased （P<0.01）.Compared with healthy group， the percentages of RANKL+ cells in plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）；compared with periodontitis group， the percentage of RANKL+ cells in the plasma cells of gingiva the patients in periodontitis+RA group was increased（P<0.05）. Compared with healthy group， the percentages of RANKL in plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）. Compared with healthy group， the number of normal induced TRAP+osteoblast-like cells of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）； compared with periodontitis group， the number of normal induced TRAP+ osteoblast-like cells of the patients in periodontitis+RA group was increased （P<0.05）； compared with the normal induced cells， the numbers of anti-RANKL induced TRAP+osteoblast-like cells in various groups were decreased（P<0.05）.Compared with healthy group，the expression levels of RANKL， RANK， and TRAF6 mRNA in the plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were significantly increased （P<0.05）；compared with periodontitis group， the expression levels of RANKL， RANK， and TRAF6 mRNA in the plasma cells of gingiva of the patients in periodontitis+RA group were increased （P<0.05）. Conclusion The differentiation level and the ability on promoting osteoclastogenesis of plasma cells of gingival of the patients with periodontitis complicated with RA are higher than those in the patients with single periodontitis， which may be a potential cause for RA aggravating periodontal tissue destruction in the patients with periodontitis.

Objective： To discuss the effect of Huaier aqueous extract （HAE） on the proliferation， migration， cell cycle，and apoptosis of tamoxifen （TAM） resistant breast cancer LCC2 cells， and to clarify its mechanism. Methods The cells were divided into control group （given DMEM basal medium） and TAM group （given 2 μmol·L^－1 TAM）， TAM+HAE group （given 2 μmol·L^－1 TAM+4 g·L^－1 HAE） and 0， 2， 4， 8， and 16 g·L^－1 HAE groups. MTT method was used to detect the survival rates and the proliferation rates of the cells in various groups； cell scratch test was used to detect the migration rates of the cells in various groups；flow cytometry was used to detect the percentages of the cells at different cell cycles and apoptotic rates of the cells in various groups；Transwell chamber test was used to detect the migration number of the cells in various groups； Western blotting method was used to detect the expression levels of estrogen receptor α （ER α） in each group，amplified in breast cancer 1（AIB1），and survivin proteins in the cells in various groups. Results The MTT assay showed that after treated for 24 h， compared with 0 g·L^－1 HAE group， the survival rates of the HAE cells in 4，8，and 16 g·L^－1 HAE groups were significantly decreased（P<0.01）. At 48 h after treatment， compared with 0 g·L^－1 HAE group， the survival rates of the cells in 2，4，8， and 16 g·L^－1 HAE groups were significantly decreased （P<0.01）；compared with 4 g·L^－1 HAE group，the survival rate of the cells in 8 g·L^－1 HAE groups was decreased（P<0.05）；compared with 24 h after treatment， the surival rates of the cells in different concentrations of HAE groups were significantly decreased （P<0.01） after treated for 48 h. At 48 treatment，compared with TAM and 4 g·L^－1 HAE group，the proliferation rate of the cells in TAM+HAE group was significantly decreased（P<0.01）.The cell scratch test results showed that the cell migration rate of the cells in 4 g·L^－1 HAE group was lower than that in 0 g·L^－1 HAE group after treated for 48 h （P<0.05）. The flow cytometry results showed that after treated for 48 h， compared with 0 g·L^－1 HAE group， the percentage of the cells at S phase and the apoptotic rate of the cells in 4 g·L^－1HAE group were significantly increased （P<0.05 or P<0.01）. The Transwell chamber experiment results showed that after treated for 48 h， compared with TAM group and 4 g·L^－1 HAE group， the migration number of the cells in TAM+HAE group was significantly decreased （P<0.01）. The Western blotting results showed that compared with MCF-7 cells，the expression levels of ERα，AIB1， and survivin proteins in the LCC2 cells were increased （P<0.05 or P<0.01）；at 48 h after treatment， compared with 0 g·L^－1 HAE group，the expression levels of ERα，AIB1， and survivin proteins in the LCC2 cells in 2，4，8， and 16 g·L^－1 HAE groups were significantly decreased （P<0.01）. Conclusion HAE can inhibit the proliferation and migration， arrest the cell cycle at S phase， and promote the apoptosis of the LCC2 cells，and it can also restore the sensitivity of the LCC2 cells to TAM， and its mechanism may be related to down-regulation of the expression levels of ER α， AIB1，and survivin proteins in the LCC2 cells.

Objective To discuss the clinical efficacy of minimally invasive circumcision， traditional open surgery， and combination surgery in the treatment of multiple breast tumors complicated with larger lesions， and to provide the clinical the basis for the treatment of the patients with multiple breast tumors. Methods A total of 167 patients with multiple breast tumors were divided into minimally invasive surgery group （n=83， treated with minimally invasive circumcision）， traditional surgery group （n=42， treated with traditional open surgery）， and combination surgery group （n=42， treated with minimally invasive circumcision and traditional open surgery）. The surgical time， intraoperative bleeding vomule， healing time of the incision， ratios of incision number to tumor number， and postoperative incidences of ecchymosis， infection， hematoma， obvious scar， and residual lesion，and the satisfaction rates of the patients in three groups were observed. Results Compared with traditional surgery group， the ages of the patients in minimally invasive surgery group and combination surgery group were younger （P<0.01）. Compared with traditional surgery group and minimally invasive surgery group，the surgical time of the patients in combination surgery group were significantly increased （P<0.01）；compared with traditional surgery group， the intraoperative bleeding vomule of the patients in combination surgery group was decreased （P<0.01）；compared with minimally invasive group， and the intraoperative bleeding vomule of the patients in combination surgery group was increased （P<0.01）.Compared with traditional surgery group and minimally invasive surgery group， the ratio of incision number to tumor number of the patients in combination surgery group were decreased significantly （P<0.01）； compared with minimally invasive surgery group，the wound healing time of the patients in combination surgery group was decreased significantly （P<0.01）. There were no statistically significant differences in the incidences of total postoperative complications， hematoma， ecchymosis， obvious scars， infection， and residual complications of the patients among three groups （P>0.05）. Compared with traditional surgery group， the psychological satisfaction rate of the patients in combination surgery group was increased （P<0.05）. Conclusion The minimally invasive breast circumcision combined with traditional open surgery have a good therapeutic effect in the treatment of the patients with multiple breast tumors complicated with large lesions，with fewer surgical incisions and high satisfaction rates；it is suitable for the promotion and application in the specific populations.

Objective To discuss the effect of Cav 3.2 gene in the treatment of stress urinary incontinence （SUI） of the mice by pelvic floor electrical stimulation （PES）， and to clarify its related mechanism. Methods Thirty C57BL/6 mice were randomly divided into control group ［without vaginal dilation （VD） modeling］， VD group （VD modeling）， and VD+PES group （VD modeling combined with PES） according to the intervention measures， and there were 10 mice in each group. The mice were divided into WT-VD group （wild-type VD modeling）， WT-VD+PES group （wild-type VD modeling combined with PES）， KO-VD group （Cav 3.2 knockout type VD modeling）， and KO-VD+PES group （Cav 3.2 knockout type VD modeling combined with PES） according to the genotype and intervention mesures. Masson staining was used to observe the deposition and expression levels of collagen type Ⅰ（Col Ⅰ） and collagen type Ⅲ （Col Ⅲ） collagen fibers in urethral and anterior vaginal wall tissue of the mice in various groups. The maximum bladder capacity（MBC） and leakage point pressure （LPP） of the mice in various groups were detected， and the expression levels of integrins β1， calpain 2， Col Ⅰ， and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in various groups were detected by Western blotting method. Results Compared with VD group， the expression levels of Col Ⅰ and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in VD+PES group mice were significantly increased （P<0.01）； compared with WT-VD group， the expression levels of Col Ⅰ and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were significantly increased （P<0.01）； compared with WT-VD+PES group， the expression levels of Col I and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in KO-VD and KO-VD+PES groups were significantly decreased （P<0.01）. Compared with control group， the MBC and LPP of the mice in VD group were decreased （P<0.05）； compared with VD group， the MBC and LPP of the mice in VD+PES group were increased （P<0.05）. The Western blotting results showed that compared with VD group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in VD+PES group were significantly increased （P<0.01）； compared with WT-VD group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were increased （P<0.01）； compared with WT-VD+PES group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissues of the mice in KO-VD and KO-VD+PES groups were significantly decreased（P<0.01）. Compared with WT-VD group， the expression levels of integrin β1 and calpain 2 proteins in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were increased significantly （P<0.01）. Conclusion PES can improve the urodynamic function of the mice and promote the generation of pelvic floor collagen and promote the repairment of pelvic floor through the Cav 3.2 gene and its downstream integrin β 1 and calpain 2.

Objective To discuss the effect of CD147 on the glycolysis of the prostate cancer （PCa） LNCaP cells， and to provide new molecular markers and targets for the clinical diagnosis and treatment of PCa. Methods The lentivirus infection system was used to establish the cells silencing CD147 as LNCaP/shCD147 group， and the negative control group （LNCaP/scramble group） was set up at the same time. The expression levels of CD147 mRNA in the LNCaP cells in two groups were detected by real-time fluorescent quantitative PCR （RT-qPCR）method； the concentrations of lactic acid （LA）， pyruvate （PA），and ATP in the LNCaP cells in two groups were detected by the kits； the activities of pyruvate kinase （PK），6-phosphofructokinase 1 （PFK1） and hexokinase （HK） were detected by the kits，and the expression levels of HK2， phosphofructokinase of muscle（PFKM），and pyruvate kinase M2（PKM2） proteins in the LNCaP cells in two groups were detected by Western blotting method. Results Compared with LNCaP/scramble group， the expression levels of CD147 mRNA and protein in the cells in LNCaP/shCD147 group were significantly decreased（P<0.01）；indicating that the PCa LNCaP cell model with silencing CD147 was successfully constructed.Compared with LNCaP/scramble group， the concentrations of LA， PA， and ATP in the cells in LNCaP/shCD147 group were significantly decreased （P<0.05 or P<0.01）. Compared with LNCaP/scramble group， the activities of PK， PFK1， and HK enzymes in the cells in LNCaP/shCD147 group were decreased （P<0.05 or P<0.01）.The Western blotting results showed that compared with LNCaP/scramble group， the expression level of HK2 protein in the cells in LNCaP/shCD147 group was decreased （P<0.01）. Conclusion Silencing CD147 inhibits the glycolysis of LNCaP cells by regulating the glycolytic metabolites of glycolysis， rate-limiting enzyme activity， and related protein expression levels.

Objective To discuss the effect of metformin on the hypertrophy of the H9C2 cardiomyocytes induced by isoproterenol （ISO）， and to clarify its possible molecular mechanism. Methods The H9C2 cells were cultured； 50 μmol·L^－1 ISO was used to establish the cardiomyocyte hypertrophy model，and they were used as ISO group；the cells without any treatment were used as control group. The expression levels of brain natriuretic peptide （BNP） in the H9C2 cardiomyocytes in two groups were detected by Western blotting method.The cells were divided into control group， ISO group， ISO+metformin group， and metformin group.The viabilities of the H9C2 cardiomyocytes in two groups were detected by CCK-8 method； the optimal drug intervention concentration and time of metformin were selected；crystal violet staining was used to observe the morphology and cross-sectional area of the H9C2 cardiomyocytes；real-time fluorescence qualitative PCR（RT-qPCR） method was used to detect the expression levels of atrial natriuretic peptide （ANP） in the cells in various groups；the morphology of mitochondriums in the H9C2 cardiomyocytes was observed by fluorescence probes； Western blotting method was used to detect the expression levels of optic atrophy 1 （OPA1） and mitofusin（MFN） proteins in the cells in various groups. Results Compared with control group，the cross-sectional area of the H9C2 cardiomyocytes in ISO group was increased significantly （P<0.05）， and the expression level of BNP protein was increased significantly （P<0.05）， suggesting that the cardiomyocyte hypertrophy model was successfully modeled. The RT-qPCR results showed that compared with control group， the expression level of ANP mRNA in the cells in ISO group was increased（P<0.05）； compared with ISO group， the expression levels of ANP mRNA in the cells in metformin+ISO and metformin groups were decreased（P<0.05）. The mitochondrium fluorescence probe observing results showed that the punctate mitochondrum in control group showed green fluorescence， mostly short rod-shaped， and a few mitochondrum were filamentously connected；compared with control group， the number of punctate mitochondrum in the H9C2 cells in ISO group was significantly increased，and the numbers of punctate mitochondrium in ISO+meformin and meformin groups were significantly decreased.The Western blotting results showed that compared with control group，the expression levels of OPA1 and MFN2 proteins in the cells in ISO group were decreased （P<0.05）；compared with ISO group， the expression levels of OPA1 and MFN2 proteins in the cells in ISO+metformin and metformin groups were increased（P<0.05）. Conclusion Metformin can alleviate the ISO-induced hypertrophy of the H9C2 cardiomyocytes，and its mechanism may be related to improving the mitochondrium fusion by up-regulating the expression levels of OPA1 and MFN2 proteins.

Objective To analyze the clinical and pathological characteristics of the mixed renal cell carcinoma （RCC） patient with co-existence of unilateral renal eosinophilic papillary renal cell carcinoma （OPRCC）and renal clear cell carcinoma （CCRCC）， and to improve the understandings of this disease. Methods The clinical data of one RCC patient with co-existence of OPRCC and CCRCC were retrospectively analyzed， and the diagnosis， treatment， and prognosis of the patient were analyzed in combination with the literature review. A 52-year-old male patient was admitted to the hospital due to a right renal mass detected by physical examination. The results of plain scan and enhancement CT of both kidneys showed space-occupying lesions of the right kidney， which was highly likely considered to be malignant. The laparoscopic partial nephrectomy was performed. Results The postoperative pathological results showed mixed RCC（lower pole of right kidney）.The results of immunohistochemical staining showed CK （AE/AE3） （partial +）， Vimentin （partial +）， EMA （partial +）， CK7 （+）， CD10 （+）， CAIX （partial +）， P504s （+）， PAX-8 （weak +）， TFE3 （－）， HMB45 （－）， MelanA （－）， SDHB （+）and Ki-67 （the positive rate was 2%）.The mixed RCC of the right kidney was diagnosed. The patient recovered quickly after operation and did not receive any adjuvant therapy. The CT examination results showed no local tumor recurrence and metastasis 3 months after operation， and no discomfort symptoms were found during the follow-up 6 months after operation. Conclusion The RCC patient with co-existence of OPRCC and CCRCC has no specific clinical manifestations， and the diagnosis mainly depends on the histopathological examination. Surgery is the first choice for the treatment；the malignant degree of the tumor is low， the progression is slow， the prognosis is good，and the long-term and close follow-up is still needed after operation.

Objective To discuss the protective effectof follistatin-like 1 （FSTL1） on doxorubicin （DOX）-induced acute myocardial injury of the mice，and to clarify its related mechanism. Methods A total of 80 C57BL/6J mice were used to establish the acute myocardial injury models by intraperitoneal injection of DOX for one time.The mice were divided into 5 groups （n=8） according to the different doses （0，5，10，15， and 20 mg·kg^－1） of DOX，and didvided into 5 groups（n=8） according to the different intervention time（0，0.5，1.0，2.0，and 3.0 d）.The other 32 mice were randonly divided into normal saline group，FSTL1 group，DOX group， and DOX-FSTL1 group.The echocardiographic and hemodynamic indexes of the mice in various groups were detected；enzyme-linked immunosorbent assay（ELISA） method was used to detect the levels of tumor necrosis factor-α（TNF-α），N-terminal pro-brain natriuretic peptide（NT-proBNP） and cardiae troponin-T（cTn-T） in serum of the mice in various groups；the activities of superoxide dismutase （SOD）， the levels of malondialdehyde （MDA）， 4-hydroxynonenal （4-HNE） and 15-F_2t isoprostane in myocardium tissue of the mice in various groups were detected by oxidative stress kit；the expression levels of FSTL1 mRNA in myocardium tissue of the mice in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method； the expression levels of nuclear factor E2 related factor 2 （Nrf2） and FSTL1 proteins in myocardium tissue of the mice in various groups were detected by Western blotting method. Results Compared with normal saline group， the left ventricular ejection fraction（LVEF），left ventricular fraction shortening（ LVFS）， left ventricular systolic pressure（LVSP）， maximum rise rate of left ventricular pressure（+dP/dt_max），and maximum drop rate of left ventricular pressure（－dp /dt_max） of the mice in DOX group and DOX+FSTL1 group were significantly decreased（P<0.05）， and the heart rate（HR），left ventricular end-diastolic volume（LVEDV），and left ventricular diastolic pressure were increased （P<0.05）； compared with DOX group， the LVEF，+dP/dt_max， and －dp /dt_max of the mice in DOX+FSTL1 group were significantly increased（P<0.05），and the LVEDV was significantly decreased （P<0.05）. Compared with normal saline group， the serum levels of TNF-α， NT-proBNP and cTn-T in serum of the mice in DOX group and DOX+FSTL1 group were significantly increased （P<0.05）； compared with DOX group， the serum levels of NT-proBNP and cTn-T in serum of the mice in DOX+FSTL1 group were significantly decreased （P<0.05）. Compared with normal saline group， the activity of SOD in myocardium tissue of the mice in DOX group was significantly decreased （P<0.05）， and the levels of MDA， 4-HNE， and 15-F_2t-isoprostane were significantly increased （P<0.05），the level of 15-F_2t-isoprostane in serum of the mice in DOX+FSTL1 group was increased （P<0.05）； compared with DOX group， the SOD activity in myocardium tissue of the mice in DOX+FSTL1 group was increased （P<0.05）， and the levels of MDA， 4-HNE， and 15-F_2t-isoprostane were decreased （P<0.05）.Compared with 0 mg·kg^－1 DOX group， the levels of FSTL1 mRNA in myocardium tissue of the mice in the other groups were significantly decreased with the increasing of the doxorubicin dose （P<0.05）. Compared with DOX 0 d group， the levels of FSTL1 mRNA in myocardium tissue of the mice in the other groups were significantly decreased with the prolongation of the doxorubicin intervention time （P<0.05）. Compared with normal saline group，the expresison levels of FSTL1 and Nrf2 proteins in myocardium tissue of the mice in DOX group was decreased（P<0.05）；compared with DOX group， the expression levels of FSTL1 and Nrf2 proteins in myocardium tissue of the mice in DOX+FSTL1 group were increased （P<0.05）. Conclusion FSTL1 can alleviate the DOX-induced acute myocardial injury and improve the cardiac function by regulating the oxidative stress， and its mechanism may be reated to up-regulating the Nrf2 expression.

Objective To discuss the clinical and pathological characteristics of IgA nephropathy （IgAN） in the children and adults， and to clarify their clinical significances. Methods The clinical and pathological data of the patients diagnosed with primary IgAN were collected. According to their ages， the patients were divided into children group （n=160） and adult group （n=240）. The ages，genders，incidences of hypertension，time from onset to renal biopsy，estimated glomerular filtration rates，initial onset manifestations，clinical types，Lee’s grades， MEST-C scores，immunofluorescence types and ultrastructures of glomeruli of the patients in two groups were compared. Results The ratio of male to female in children group was 1.5∶1 and it was 1.1∶1 in adults group； compared with adult group， the incidence of hypertension and the time from onset to renal biopsy of the patients in child group were decreased （P<0.05）， while the eGFR was increased（P<0.05）. Compared with adult group， the percentages of gross hematuria and edema of the patients in child group were increased（P<0.01）， and the percentages of abnormal urine test， foam urine， and other manifestations were decreased（P<0.01）.Compared with adult group， the percentages of the patients with isolated hematuria and nephrotic syndrome in child group were increased （P<0.01）， while the percentages of the patients with isolated proteinuria and chronic nephritis were decreased （P<0.01）；compared with adult group， the percentage of the patients with grade Ⅱ in the Lee’s grades in child group was increased（P<0.01） and the percentage of the patients with grade Ⅴ in the Lee’s grades was decreased （P<0.01）.The light microscope obervation results showed that there were only focal mesangial cells and mesangial matrix hyperplasia in grade Ⅱ renal tissue of the patients in child group， rarely accompanied by glomerulosclerosis. The renal tubular and interstitial lesions were not significant； there were significant proliferations of the mesangial cells and matrix in grade Ⅴ renal tissue of the patients in adult group， with more glomerulosclerosis， and relatively severe renal tubular and interstitial lesions. Compared with adult group， the percentages of the patients with M1 and E1 in the MEST-C scores of the patients in child group were increased （P<0.05 or P<0.01）， and the percentages of the patients with S1 and T1/T2 were decreased （P<0.01）.The urinary protein grade of the patients in child group was positively correlated with M（r_s=0.462），E（r_s=0.342），and C（r_s=0.250）scores （P<0.01）；the urinary protein grade of the patients in adult group was positively correlated with M（r_s=0.217），E（r_s=0.145），S（r_s=0.187），T（r_s=0.269），and C（r_s=0.256）scores （P<0.01）；compared with adult group， the IgA+IgG+IgM deposition of the patients in child group was increased（P<0.05）， deposition rate of C3 was increased（P<0.01）， and the deposition rate of fibrinogen （Fib） was increased（P<0.01）. Conclusion There are significant differences in the clinical and pathological characteristics between the children and the adults with primary IgAN， which should be treated differently in the clinical diagnosis， treatment， and research.

Objective To discuss the protective effect of Yiyi Fuzisan on the myocardium ischemia and vascular endothelial dysfunction of the mice， and to clarify its possible mechanism. Methods Sixty male SPF grade C57BL/6J mice were randomly divided into blank group， sham operation group， acute myocardial ischemia （AMI） group， and low，medium and high doses of Yiyi Fuzisan groups， and there were 10 mice in each group；another forty C57BL/6J mice were randomly divided into normal saline group and low， medium， and high doses of Yiyi Fuzisan groups， and there were 10 mice in each group. The left anterior descending coronary artery （LAD） ligation method was used to establish the AMI mouse model， and corresponding drug intervention treatment was performed for 28 d. Western blotting method was used to detect the expression levels of endothelial nitric oxide synthase （eNOS） protein in myocardium tissue of the mice in various groups；arterial tension detection method was used to detect the tension of isolated aortic and relaxation rates of the mice in various groups；total nitric oxide（NO） detection kit was used to detect the levels of NO in serum of the mice in various groups；triphenyltetra zolium chloride（TTC） staining was used to observe the ischemia areas of myocardium tissue of the mice in various groups； HE staining was used to observe the pathomorphology of myocardium tissue of the mice in various groups；the postoperative survival rates of the mice in various groups were observed. The human coronary endothelial cells （HCAECs） were randomly divided into blank group， hypoxia group，hypoxia+ low dose of Yiyi Fuzisan group， hypoxia+medium dose of Yiyi Fuzisan group， and hypoxia+high dose of Yiyi Fuzisan group. Except for blank group， the cells in the other groups were cultured in the three gas incubator for 24 h to establish the hypoxia models. Griess test was used to detect the levels of NO in the HCAECs in various groups；fluorescence staining was used to detect the levels of mitochondrial membrane potential （MMP） and reactive oxygen species （ROS） in the HCAECs in various groups. Results Compared with before establishing the AMI model， the ST segment of electrocardiogram of the mice was significantly elevated after establishing the AMI model. The Western blotting results showed that compared with blank group， the expression levels of eNOS protein in myocardium tissue of the mice in sham operation group， AMI group，and low， medium， and high doses of Yiyi Fuzisan groups were decreased after injection of N-nitro-L-arginine-methylesterhydro chloride（L-NAME）（P<0.05）. Compared with saline group， the relaxation rates of thoracic aorta of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased （P<0.05）. Compared with blank group and sham operation group， the serum NO level of the mice in AMI group was decreased （P<0.05）， the serum NO level of the mice in high dose of Yiyi Fuzisan group was increased （P<0.05）； compared with AMI group， the serum NO levels of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased （P<0.05）. The TTC staining observation showed that compared with blank group and sham operation group， there were various degrees of myocardium ischemia of the mice in myocardium tissue of the mice in AMI group and low， medium， and high doses of Yiyi Fuzisan groups； compared with AMI group，the ischemia areas of myocardium tissue of the mice in low， medium， and high doses of Yiyi Fuzisan groups were decreased （P<0.05），and the percentages of ischemia areas of myocardium tissue were decreased.The HE staining results showed that the cardiomyocytes of the mice in blank group and sham operation group were neatly arranged， with clear and complete nuclei and uniform sizes，and no inflammatory cell infiltration was observed；the cardiomyocytes of the mice in AMI group showed significant myocardial tissue damage， with disordered arrangement，rupture and necrosis of cardiomyocytes， and infiltration of inflammatory cells；the myocardium tissue of the mice in low， medium， and high doses of Yiyi Fuzisan groups showed pathological recovery of myocardial tissue damage. On the 28th day after drug intervention， the survival rates of the mice in blank group and sham operation group were 100%； compared with AMI model group， the survival rates of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased（χ²=15.03，P=0.0102）.The Griess experiment results showed that compared with blank group， the levels of NO in the HEAEs in hypoxia group and hypoxia+low dose of Yiyi Fuzisan group were decreased （P<0.05）； compared with hypoxia group， the levels of NO in the HEAECs in hypoxia+medium dose of Yiyi Fuzisan and hypoxia+high dose of Yiyi Fuzisan groups were increased （P<0.05）. The fluorescence staining results showed that compared with blank group， the levels of MMP in the cells in hypoxia group， hypoxia group+low dose of Yiyi Fuzisan group， and hypoxia +medium dose of Yiyi Fuzisan group were decreased （P<0.05），while the levels of ROS were increased （P<0.05）； compared with hypoxia group， the levels of MMP in the HEAECs in hypoxia+low dose of Yiyi Fuzisan，hypoxia+medium dose of Yiyi Fuzisan，and hypoxia+high dose of Yiyi Fuzisan groups were increased （P<0.05）， while the levels of ROS in the HCAECs in hypoxia+low dose of Yiyi Fuzisan，hypoxia+medium dose of Yiyi Fuzisan，and hypoxia+high dose of Yiyi fuzisan groups were decreased （P<0.05）. Conclusion Yiyi Fuzisan can improve the pathological state of myocardium ischemia， restore the MMP， reduce the production of ROS， and improve the dysfunction of mitochondria and vascular endothelial cells by enhancing the biological activity of NO and increasing the vascular activity of aorta.

Objective To investigate the effect of paired box transcription factor 3 （PAX3） gene silencing on the differentiation of the P19 cells into the cardiomyocyte-like cells，and to clarify its possible mechanism. Methods Dimethylene maple （DMSO） was used to induce the differentiation of the P19 cells into cardiomyocyte-like cells， and the morphology of the cells during the induction process was observed. The cells were collected on the 0th day （0 d group） and 10th day （10 d group） after induction and differentiation.The P19 cells were infected with sh-PAX3 Lentivirus.The cells were divided into control group，sh-NC group（negative control group）and sh-PAX3 （PAX3 interference）.Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to verify the positive expression of cTnI in P19 cells silenced with PAX3 gene. The P19 cells infected with lentivirus were treated with Notch signal agonist Jagged1 and were induced and differentiated with DMSO for 10 d. The cells were divided into DMSO group， DMSO+sh-NC group， DMSO+sh-PAX3 group， DMSO+Jagged1 group and DMSO+sh-PAX3+Jagged1 group. RT-qPCR method was used to detect the expression levels of GATA binding protein 4 （GATA4）， atrial natriuretic peptide （ANP）， cardiac troponin （cTn）T，PAX3，Notch1， Notch intracellular domain （NICD）， and Hes family bHLH transcription factor 1 （Hes1） mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of PAX3，Notch1， NICD， and Hes1 proteins in the cells in various groups； immunofluorescence assay was used to detect the positive expression of cTnI in the cells the differentiation rates of the cardiomyocyte-like cells in various groups. Results On the 10th day， after induction and differentiation，the cluster of cardiomyocyte-like cells with spontaneous rhythmic contraction could be observed.The RT-qPCR and Western blotting results showed that after lentivirus infection， compared with control group and sh-NC group， the expression levels of PAX3 mRNA and protein in the P19 cells in sh-PAX3 group were decreased （P<0.05）， indicating that the P19 cells silenced with PAX3 gene were successfully constructed. The RT-qPCR results showed that compared with 0 d group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in 10 d group were increased （P<0.05）， while the expression levels of PAX3，Notch1， NICD， and Hes1 mRNA were increased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+sh-PAX3 group were decreased （P<0.05）， while the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+Jagged1 group were increased （P<0.05）； compared with DMSO+sh-PAX3 group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+sh-PAX3+Jagged1 group were increased （P<0.05）. The Western blotting results showed that compared with 0 d group， the expression levels of PAX3， Notch1， NICD， and Hes1 proteins in the cells in 10 d group were increased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+sh-PAX3 group were decreased （P<0.05），while the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+Jagged1 group were increased （P<0.05）； compared with DMSO+sh-PAX3 group， the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+sh-PAX3+Jagged1 group were increased （P<0.05）.The immunofluorescence results showed that after lentivirus infection， the positive expression of cTnI protein was found in the cells in various groups. Compared with DMSO group and DMSO+sh-NC group， the positive expression of cTnI protein in the cells in DMSO+sh-PAX3 group was decreased， and the differentiation rate of the cardiomyocyte-like cells was decreased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the positive expression of cTnI protein in the cells in DMSO+Jagged1 group was increased，and the differentiation rate of the cardiomyocyte-like cells was increased （P<0.05）； compared with DMSO+sh-PAX3 group， the positive expression of cTnI protein in the cells in DMSO+sh-PAX3+Jagged1 group was increased，and the differentiation rate of the cardiomyocyte-like cells was increased （P<0.05）. Conclusion PAX3 gene silencing may inhibit the differentiation of the P19 cells into the cardiomyocyte-like，and its mechanism may be realized by decreasing the PAX3 gene expression and regulating the Notch signaling pathway.

Objective To analyze the changes of cholesterol levels of the patients with acute coronary syndrome （ACS）， and to explore the predictive value of residual cholesterol （RC） levels in the occurrence of ACS. Methods A total of 220 ACS patients were selected as ACS group； 220 healthy people underwent physical examination at the same time were selected as control group. The general informations including serum levels of total cholesterol （TC）， low density lipoprotein cholesterol （LDL-c） and high density lipoprotein cholesterol （HDL-c） of the subjects in two groups were collected， and the levels of serum non-high density lipoprotein cholesterol （non HDL-c） and serum residual cholesterol （RC） were calculated. Logistic regression analysis was used to analyze the risk factors of the ACS patients and their correlations with the levels of various lipid components. The receiver operating characteristic curve（ROC） and the area under （AUC） were used to evalue the predictive values of various blood lipid components in the ACS. Results The single factor analysis results showed that compared with control group，the percentages of the patients with hypertension and smoking in ACS group were increased（P<0.01），the levels of TC， RC， LDL-c，non HDL-c of the patients in ACS group were increased（P<0.01），and the level of HDL-c was decreased（P<0.01）.The multivariate Logistic regression analysis results showed that hypertension （OR=0.496， P=0.005）， smoking （OR=0.458， P=0.002），RC level （OR=24.051，P<0.01）， and non HDL-c level（OR=1.711， P<0.01） were the independent risk factors for the occurrence of ACS. The ROC curve analysis results show that compared with non-HDL-c level， the AUC of RC level was larger，which had the higher predictive value for the occurrence of the ACS patients. Conclusion RC level is an independent risk factor for the occurrence of ACS， and has certain predictive value for the occurrence of ACS.

Objective To discuss the regulatory effect of long chain non-coding RNA （lncRNA） metastasis associated transcript 1 （MALAT1） on the activation of the hepatic stellate cells （HSC） during the progression of liver fibrosis， and to clarify its mechanism. Methods The serum samples were collected from 25 healthy volunteers （n=25，healthy group） and 25 liver fibrosis patients［ mild liver fibrosis group （n=12）and severe liver fibrosis group （n=13）］. The mouse HSC were divided into control group，transforming growth factor-β1（TGF-β1） group， TGF-β1+si-NC group， TGF-β1+si-MALAT1 group，TGF- β 1+si-MALAT1+anti-miR-150-5p group，and TGF-β1 group+si-MALAT1+CXCL14 group. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of MALAT1 mRNA， microRNA-150-5p （miR-150-5p）， and CXC chemokine ligand 14 （CXCL14） mRNA in serum and HSC of the subjects in various groups；Western blotting method was used to detect the expression levels of CXCL14， α-smooth muscle actin（α-SMA），and type Ⅰ collagen α 1 （COL1A1） proteins in serum of the subjects in various groups；CCK-8 method was used to detect the proliferation activities of the HSC in various groups； the expression levels of α- SMA，COL1A1 proteins in the HSC in various groups were detected by immunofluorescence；the targeting relationship between miR-150-5p and MALAT1 and CXCL14 3′-UTR genes was detected by dual luciferase reporting system. Results .The RT-qPCR results showed that compared with healthy group， the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in mild and severe liver fibrosis groups were increased（P<0.05）， while the expression levels of miR-150-5p were decreased （P<0.05）； compared with mild liver fibrosis group， the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in severe liver fibrosis group were increased （P<0.05），while the expression level of miR-150-5p was decreased （P<0.05）； compared with control group， the expression levels of MALAT1 mRNA and CXCL14 mRNA in the HSC in TGF- β1 group were increased （P<0.05）， while the expression level of miR-150-5p was decreased （P<0.05）； compared with TGF-β1+si-NC group， the expression levels of MALAT1 mRNA and CXCL14 mRNA in the HSC in TGF-β1+si MALAT1 group were decreased （P<0.05）， while the expression level of miR-150-5p was increased （P<0.05）； compared with TGF-β1+si MALAT1 group， the expression levels of miR-150-5p in the HSC in TGF- β1+si-MALAT1 and anti-miR-150-5p groups were decreased （P<0.05）， while the expression levels of CXCL14 mRNA were increased （P<0.05）； compared with TGF-β1+si-MALAT1 group， the expression level of CXCL14 mRNA in the HSC in TGF-β1+si-MALAT1+CXCL14 group was increased （P<0.05）.The Western blotting results showed that compared with healthy group， the serum expression levels of CXCL14 protein of the subjects in mild and severe liver fibrosis groups were increased （P<0.05）； compared with mild liver fibrosis group， the serum expression level of CXCL14 protein of the subjects in severe liver fibrosis group was increased （P<0.05）； compared with control group， the expression levels of α-SMA and COL1A1 proteins in the HSC in TGF-β1 group were increased （P<0.05）； compared with TGF-β1+si-NC group， the expression levels of α- SMA and COL1A1 proteins in the HSC in TGF-β1+si-MALAT1 group were decreased （P<0.05）； compared with TGF-β1+si-MALAT1 group， the expression levels of CXCL14，α-SMA and COL1A1 proteins of the HSC in TGF-β1+si-MALAT1+anti-miR-150-5p group and TGF-β1+siMALAT1+CXCL14 group were increased （P<0.05）.The CCK-8 method results showed that compared with control group， the proliferation activity of the HSC in TGF-β1 group was increased （P<0.05）； compared with TGF-β1+si-NC group， the proliferation activity of the HSC in TGF-β1+si-MALAT1 group was decreased （P<0.05）； compared with TGF-β1+si-MALAT1 group，the proliferation activities of the HSC in TGF-β1+si-MALAT1+anti-miR-150-5p group and TGF-β1+si-MALAT1+CXCL14 group were increased （P<0.05）. The immunofluorescence results showed that the expressions of α-SMA and COL1A1 proteins in the HSC in various groups was consistent with the results detected by Western blotting method. There was a targeting relationship between MALAT1 and CXCL14 3 '-UTR and miR-150-5p. The double luciferase reporter gene assay results showed that compared with miR-NC group， the luciferase activity of the HSC in miR-150-5p group co-transfected with MALAT1 WT or CXCL14 WT was decreased （P<0.05）. Conclusion Knocking down of MALAT1 can inhibit the activation of the HSC induced by TGF- β 1，and the mechanism may be related to the miR-150-5p/CXCL14 signaling pathway.

Objective To discuss the diagnosis and treatment of one patient with ruptured intracranial aneurysm complicated with acute myocardial infarction（AMI）， and to provide the reference for the clinical diagnosis， treatment， and anesthesia of the disease. Methods The clinical data， imaging findings， and anesthesia methods of one patient with ruptured intracranial aneurysm complicated with AMI were retrospectively analyzed，and the analysis was performed combined with the relevant literatures. Results The patient was admitted to hospital due to sudden severe headache with nausea and vomiting for 4 h. A total of 1 h and 10 min after admission，the multi-slice CT results showed subarachnoid hemorrhage（SAH）.There was little bilateral ventricular effusion， and the intracranial angiography results showed a tumor in the posterior communicating segment of the right internal carotid artery.A total of 2 h 52 min after admission，the myoglobin was 483.6 μg·L^－1，the troponin I was 4.990 μg·L^－1，the creatine kinase isoenzyme-MB（CK-MB） was 45.70 μg·L^－1.A total of 16 h 31 min after admission， the ECG results showed sinus bradycardia， left ventricular hypertrophy， and ST-T segment changes. The initial diagnosis of the patient was SAH， AMI， and hypertension grade 3 （very high risk）. After early comprehensive treatment， the patient underwent emergency clipping of cerebral aneurysm after 3 d.The anesthesia method was tracheal intubation ansthesia，and the anesthetic drugs were carefully selected to achieve the best blood flow and anesthesia effect.The vital signs of the patient were stable during the operation， and the condition of the patient was improved and discharged after 7 d. Conclusion For the patients with intracranial aneurysm rupture complicated with AMI，CT， intracranial angiography， and myocardial markers are the important examinations for the diagnosis and differential diagnosis； controlling the blood pressure is the key point for the treatment and anesthesia.

Objective To discuss the effect of ghrelin on the differentiation of the adipose mesenchymal stem cells （ADSCs） into the neurons，and to clarify its possible mechanism. Methods The ADSCs were randomly divided into blank group， neural differentiation inducer group， and ghrelin group （given 600 μg·L^－1 Ghrelin）， U0126 group （given 40 ng·L^－1 U0126）， Ghrelin+U0126 group （given 600 μg·L^－1 Ghrelin+40 ng·L^－1 U0126），and Ghrelin receptor blocker D-Lys3-GHRP-6 （D-Lys3-GHRP-6） group （given 10^－10 g·L^－1 D-Lys3-GHRP-6）. The pathomorphology of the cells in various groups was observed under inverted microscope；the positive expression rates of neurofilament protein （NF）-200 and tubulin （Tuj-1） in the cells in various groups were detected by immunofluorescence method； the expression levels of the mitogen activated protein kinase （MAPK）/extracellular signal-regulated kinase （ERK） pathway proteins，neuron specific nuclear protein （NeuN）， Tuj-1， neuron specific enolase （NSE），NF，growth hormone secretagogne receptor（GHSR），fetal liver kinase 1（Flk1）， and CD29 proteins in the cells in various groups were detected by Western blotting method. Results On the 10th day after cell culture， the ADSCs in blank group showed the fusions of long spindle and spiral shapes； the body of the cells in neural differentiation inducer group contracted，and the long spindle like cell body was transformed into the round-like shape， the protrusion length extended， and the number of neuron-like cells was increased； in Ghrelin group， the number of body protrusions of the cells was increased and the number of round-like like cells was increased； there was a small amount of cell body contraction and circular transformation in U0126 group and D-Lys3-GHRP-6 group. The immunofluorescence assay results showed that compared with blank group， the positive expression rates of NF-200 and Tuj-1 in the cells in neural differentiation inducer group were increased （P<0.05）； compared with neural differentiation inducer group， the positive expression rates of NF-200 and Tuj-1 in the cells in Ghrelin group were increased （P<0.05）， while the positive expression rates of NF-200 and Tuj-1 in the cells in U0126 group and D-Lys3-GHRP-6 group were decreased （P<0.05）； compared with Ghrelin group， the positive expression rates of NF-200 and Tuj-1 in the cells in U0126 group， D-Lys3-GHRP-6 group， and Ghrelin+U0126 group were decreased （P<0.05）. The Western blotting results showed that compared with blank group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins，and the ratios of phosphorylated MAPK（p-MAPK）/MAPK and phosphorylated ERK（p-ERK）/ERK in the cells in neural differentiation inducer group were increased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were decreased （P<0.05）； compared with neural differentiation inducer group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in Ghrelin group were increased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were decreased （P<0.05）； the expression levels of NSE， NeuN， Tuj-1， NF， and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in U0126 and D-Lys3-GHRP-6 groups were decreased （P<0.05），and the expression levels of Flk1 and CD29 proteins were increased （P<0.05）； compared with Ghrelin group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in U0126 group， D-Lys3-GHRP-6 group， and Ghrelin+U0126 group were decreased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were increased （P<0.05）. Conclusion Ghrelin can induce the differention of the ASCs into the neurons，and it mechanism is related to activating the MAPK/ERK pathway.

Objective To discuss the effect of long non-coding RNA （lncRNA） HAND2-AS1 in endometrium tissue of the patients with endometriosis （EMT） on the proliferation， migration and invasion of the endometrial stromal cells （ESCs） in ectopic endometrium（EC）tissue，and to clarify the possible mechanism. Methods The EC tissue of 30 patients with EMT （EMT group） and the endometrium tissue of 30 healthy women of childbearing age （control group） were collected；the endometrium tissue was isolated，and the ESCs were collected. The ESCs in EC tissue of the EMT patients were transfected with liposome transfection method，and the ESCs were divided into pcDNA group （transfected with pcDNA empty plasmid），HAND2-AS1 group（transfected with HAND2-AS1 over-expression plasmid），mimic NC group（transfected with mimic NC），miR-21 mimic group（transfected with miR-21 mimic），pcDNA+mimic-NC group（transfected with pcDNA and mimic NC）， HAND2-AS1+mimic NC group（transfected with HAND2-AS1 over-expression plasmid and mimic NC）， pcDNA+miR-21 mimic group （transfected with pcDNA empty plasmid and miR-21 mimic），and HAND2-AS1+miR-21 mimic group（transfected with HAND2 AS1 over-expression plasmid and miR-21 mimic），and the cells without any transfection were used as blank control group.The expression levels of HAND2-AS1 mRNA and miR-21 in the endometrium tissue and ESCs of the subjects in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method；and the relationship between them was analyzed by Pearson correlation coefficient；bioinformatics software Starbase was used to predict the targeting relationship between lncRNA HAND2-AS1 and miR-21；the luciferase gene reporting assay was used to verify.The proliferation activities of the ESCs in various groups were detected by CCK-8 method；the numbers of migration and invasion cells were detected by Transwell chamber assay. Results There were no significant differences in the age， body mass index （BMI）， serum levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH） and prolactin （PRL） of the subjects between two groups （P<0.05）. Compared with control group， the serum estradiol （E2） level of the patients in EMT group was increased （P<0.05）， the expression level of HAND2-AS1 mRNA in EC tissue of the patients in EMT group was decreased （P<0.05）， while the expression level of miR-21 was increased （P<0.05）. Compared with control group， the expression level of HAND2-AS1 mRNA in the ESCs in EMT group was decreased （P<0.05）， while the expression level of miR-21 was increased （P<0.01）.The Pearson correlation coefficient analysis results showed that there was no significant correlation between the expressions of HAND2-AS1 and miR-21 in the endometrium tissue and ESCs of the subjects in control group （r=0.34， P>0.05）， while there was a negative correlation between the expressions of HAND2-AS1 and miR-21 in the EC tissue and ESCs of the patients in EMT group （r=－0.57， P<0.05）. The RT-qPCR results showed that compared with blank control group， the expression level of HAND2-AS1 mRNA in the ESCs in HAND2-AS1 group was significantly increased（P<0.01），the miR-21 expression level in the ESCs in miR-21 mimic group was increased（P<0.01），the expression level of miR-21 in the ESCs in HAND2-AS1+mimic NC group was significantly decreased（P<0.05），and the miR-21 expression level in the ESCs in pcDNA+miR-21 mimic group was significantly increased（P<0.01）； compared with pcDNA+ miR-21 mimic group， the miR-21 expression level in the ESCs in HAND2-AS1+miR-21 mimic group was significantly decreased（P<0.05）.There was a potential binding site between lncRNA HAND2-AS1 and miR-21.The dual luciferase gene reporter assay results showed that compared with mimic NC group， the luciferase activity in HAND2-AS1-WT in miR-21 mimic group was significantly decreased（P<0.01）. Compared with blank control group， the proliferation activity of the ESCs， the numbers of migration and invasion ESCs in HAND2-AS1+mimic-NC group were significantly decreased （P<0.05 or P<0.01）， and the above indexes in pcDNA+miR-21 mimic group were significantly increased（P<0.05）； compared with pcDNA+miR-21 mimic group， the proliferation activity，the numbers of migration and invasion ESCs in HAND2-AS1+miR-21 mimic group were significantly decreased（P<0.05）. Conclusion The expressions of HAND2-AS1 in the EC tissue and ESCs of the EMT patients are significantly decreased， and it can down-regulate the proliferation， migration and invasion of the ESCs in EC tissue by targetly inhibiting the expression of miR-21， thus participats in the occurrence and development of EMT.

Objective To discuss the protective effect of ginsenoside Rh2 on kidney injury in the rats with diabetic kidney lesions （DKL），and to clarify its mechanism. Methods The DKL rat model was established by using the high-fat and high-energy diet combined with intraperitoneal injection of streptozotocin（STZ）.Thirty SD model rats were randomly divided into model group， low dose （10 mg·kg^－1） of Rh2 group and high dose （20 mg·kg^－1） of Rh2 group， and there were 10 rats in each group；other ten SD rats were regared as normal control group.The body weights and kidney weights of the rats in various groups were detected，and the kidney index was caculated；after intervented for 8 weeks， automatic analyzer was used to detect the levels of serum creatinine （Scr）， urea nitrogen （BUN） and 24 h urinary protein （UP）in serum of the rats in various groups；enzyme linked immunosorbent assay （ELISA） method was used to detect the levels of collagen type Ⅳ（Col Ⅳ） in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of transforming growth factor-β1 （TGF-β1），Smad3 and Smad7 proteins in kidney tissue of the rats in various groups. Results Compared with normal control group， the body weight of the rats in model group was decreased（P<0.01），the kidney weight and kidney index were significantly increased （P<0.01）， the levels of Scr， BUN and 24 h UP in serum were increased （P<0.01）， the level of Col Ⅳ in kidney tissue was increased（P<0.01）， the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were increased（P<0.01）， and the expression level of Smad7 protein in renal tissue was decreased （P<0.01）. Compared with model group，the body weight of the rats in low and high doses of Rh2 groups were inecreased （P<0.05 or P<0.01）， the kidney weight and kidney index were decreased （P<0.05 or P<0.01），the levels of Scr， BUN and 24 h UP in serum were significantly decreased （P<0.05 or P<0.01）， the level of Col Ⅳ in kidney tissue was decreased（P<0.01）， the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were decreased（P<0.01）， and the expression level of Smad7 protein in kidney tissue was increased （P<0.01）. Conclusion Ginsenoside Rh2 can inhibit the kidney fibrosis and improves the kidney function in the DKL rats， and has a protective effect on the kidney of DKL rats； its mechanism may be related to regulating the expression of TGF-β1/Smads signaling pathway.

Objective： To analyze the effective chemical components and their action targets of epimedium in the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis functional damage by using network pharmacology and molecular docking technology， and to preliminarily clarify its mechanism. Methods The effective chemical components and their corresponding target proteins of epimedium were screened by Traditional Chinese Medicine Systems Pharmacology （TCMSP） Database and Analysis Platform combined with the relevant literatures， and the Uniprot Database was used to standardize the target protein informations to get the corresponding standard gene names； the relevant target genes of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage were collected from the GeneCards Database and DisGeNET Database；the network of the relationship between the effective chemical components of the epimedium and their action targets was constructed by Cytascape 3.7.1 Software； the protein-protein interaction （PPI） network diagram of drug-disease of effective chemical components and target proteins of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage was constructed by STRING Database. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genome （KEGG） signaling pathway enrichment analysis were performed by OmicShare Database；molecular simulation docking and visualization were done by Pymol Software and the Autodock Tools Software. Results A total of 27 effective chemical components and 217 corresponding action targets of epimedium， and 465 target genes of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage were screened out， of which 62 target genes were closely related to the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage by epimedium.Quercetin，luteolin，kaempferol， and icariin and so on were the key effective chemical components， and the key target proteins were Cysteine protease 3 containing cysteine（Caspase-3）， interleukin-6（IL-6）， oligosaccharides （FOS）， tumor necrosis factor（TNF）， hypoxia inducible factor-1 alpha （HIF1A）， vascular endothelial growth factor A（VEGFA）， interleukin-1β（IL-1β）， estrogen receptor 1 （ESR1），recombinant prostaglandin endoperoxide synthase 2（PTGS2）， matric metallopeptidase 9（MMP-9），and so on.The KEGG signaling pathway enrichment analysis results showed that the common targets were mainly concentrated in the pathways related to immune inflammation， hormone regulation， and energy metabolism. The molecular docking results showed that the potential active components of epimedium had a good affinity with IL-6， PTGS2， aldose reductase（AR）， and mitogen-activated protein kinase 1（MAPK1）targets. Conclusion Epimedium may play a potential therapeutic role in the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage in the aspects of anti-inflammatory immunity， hormone regulation， and energy metabolism through multiple targets and pathways.

Objective To discuss the expression levels of the cyclin-dependent kinase 5 regulatory subunit associated protein 1-like 1（CDKAL1） gene and its splice isoforms in peripheral blood lymphocytes of the patients with type 2 diabetes mellitus （T2DM）， and to clarify their clinical significances. Methods A total of 65 diabetes mellitus patients were enrolled， including 20 T2DM patients （T2DM group）， 23 diabetic nephropathy（DN） patients （DN group）， 22 diabetic retinopathy（DR）patients （DR group）， and 28 healthy examiners at the same term（healthy control group）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of CDKAL1 gene and its two splice isoforms， CDKAL1-X1 and CDKAL1-X2， in peripheral blood lymphocytes of the subjects in various groups. The relationships between the clinical significances of their expressions and T2DM and its microvascular complications were analyzed. Results Compared with healthy control group， the expression levels of CDKAL1 gene （Z=4.705， P<0.01） and CDKAL1-X1 （Z=2.698， P=0.007） in peripheral blood lymphocytes of the patients in T2DM group were increased. Compared with healthy control group， the expression levels of CDKAL1 gene and CDKAL1-X1 in peripheral blood lymphocytes of the patients in DR and DN groups were increased（P<0.05）； the expression level of CDKAL1 gene in peripheral blood lymphocytes of the patients in DR group was higher than that in DN group （P<0.05）. Conclusion The high expressions of CDKAL1 gene and CDKAL1-X1 in peripheral blood lymphocytes of the T2DM patients may play an important role in the occurence and development of diabetic mellitus and its complications.

Objective To compare the clinical efficacies of transurethral plasmakinetic enucleation of prostate （PKEP） and transurethral plasmakinetic resection of prostate （PKRP） in the treatment of benign prostatic hyperplasia （BPH）， and provide the evidence for the treatment options for BPH. Methods The clincal data of 60 patients with lower urinary tract symptoms （LUTS） and initial diagnosis as BPH were collected. With the patients’ informed consent， they were divided into PKEP group （treated with PKEP） and PKRP group（treated with PKRP）， and there were 30 cases in each group. The age， body weight， body mass index （BMI）， total prostate-specific antigen （tPSA） level， hematocrit （HCT）， hemoglobin （Hb） level，preoperative and postoperative sodium ion level， prostate volume，degree of midlobe median lobe protrusion， occurrence of complications （hypertension， diabetes， and respiratory diseases）， percentage of oral 5α-reductase inhibitor using， operation time，amount of intraoperative blood， resected tissue volume， resection rate，resection efficiency，total hospital stay after operation，indwelling catheterization time， bladder irrigation duration， sodium ion level，Quality Of Life （QOL） score， and International Prostate Symptom Score （IPSS） before and after operation of the patients in two groups were observed and compared.The incidences of adverse reactions of the patients in two groups after operation were also compared. Results There were no significant differences in age， body weight， BMI，tPSA level， preoperative HCT， preoperative Hb level，sodium ion level before operation， prostate volume， and degree of midlobe protrusion of the patients between two groups （P>0.05）. There were no statistically significant differences in the occurrences of complications （hypertension， diabetes， and respiratory diseases） and the percentage of oral 5α-reductase inhibitor using of the patients between two groups （P>0.05）； the amount of blood during operation， resected tissue volume， resection rate， and resection efficiency of the patients in PKRP group were higher than those in PKEP group （P<0.05）； there were no significant differences in the total hospital stay，indwelling catheterization time， and bladder irrigation duration after operation of the patients between two groups （P>0.05）； there were no significant differences in the sodium ion levels， QOL scores， and IPSS of the patients between two groups before and after operation（P>0.05）； there were significant differences in the QOL scores and IPSS of the patients in each group before and after operation （P<0.05）.There was one case of urinary incontinence in PKEP group and two cases of urinary incontinence in PKRP group， without other operation-related complications. Conclusion Both two methods have similar efficacy and safety， and can achieve the satisfactory surgical outcomes.The patients undergoing PKEP have less amount of blood during operation， higher resected tissue volume，the higher resection efficiency， and higher resection rate， which is a better way to treat BPH.

Objective To seek the prognostic markers for the gastric cancer （GC） patients， and to achieve the early diagnosis and treatment of GC， and to provide the evidence for improving the survival rate of the GC patients. Methods The clinical data and transcriptome data of 407 and 433 GC patients were downloaded from the The Cancer Genome Atlas（TCGA） Database and GSE84437 Dataset， and merged； the GC tissue samples were classified and typed based on the expression levels of cell death-related genes and the ConsensusClusterPlus Package，and were divided into type A（342 cases） and type B（465 cases）；the GC patients were divided into low expression group and high expression group according to the expression level of apolipoprotein D（APOD）；the survminer Data Package was used to compare the differences in prognosis of the patients with different subtypes；the ssGSEA Algorithm was used to compare the differences in immune cell infiltration of the patients with different subtypes； Lasso regression and Cox regression were used to construct the prognostic risk model for the GC patients； the clinical characteristics of model genes were filtered；the differential expression of APOD in adjacent normal tissue and GC tissue was analyzed by Public Databases；GSEA analysis was used to assess the Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment of APOD； the CIBERSORT Algorithm was used to evaluate the correlation between the content of immune cells and expression of APOD；the online website was used to analyze the drug sensitivity of APOD. Results There was statistically significant difference in prognosis of the patients with two subtypes of cell pyroptosis（P=0.002）； the levels of 22 kinds of immune cells of the patients with type A were higher than those with type B（P<0.01）； there were statistically significant differences in the percentages of two subtypes of cell pyroptosis of the patients with different ages （P<0.01） and N stages （P=0.04）. The Public Databases analysis results showed that the expression of APOD in tumor tissue of the GC patients was lower than that in adjacent normal tissue； the survival results showed that compared with low expression group，the patients in high expression group had poorer prognosis； the clinical correlation analysis results showed that there were significant differences in the APOD expression of the patients with different T stage （P<0.05）； the GSEA analysis results showed that high expression group enriched in the cell adhesion pathways， leukocyte endothelial migration pathways， and gap junction pathways； the immune infiltration analysis results showed that the contents of follicular helper T lymphocytes， CD4+ memory activated T lymphocytes， resting NK cells， and neutrophils in high expression group were lower than those in low expression group； APOD had positive correlations with CXC chemokine ligand 12（CXCL12）（r=0.500，P<0.01），transforming growth factor beta 1（TGFB1） （r=0.313，P<0.01），chemokine CC chemokine ligand 19（CCL19）（r=0.518，P<0.01），and CX3C chemokine receptor 1（CX3CR1）（r=0.444，P<0.01）；the drug sensitivity analysis results showed that there were positive correlations between APOD and AZD-7762， KW-2449， and TG-101348（0<r<0.3，P<0.05）. Conclusion Cell pyroptosis subtypes can effectively evaluate the prognosis of the GC patients； APOD can be regarded as the novel prognostic marker and therapeutic target for GC.

Objective To discuss the effect of paeonol on the proliferation， migration and chemokinereceptor4 （CXCR4）/signal transducer and activators of transcription3 （STAT3） pathway of the human osteosarcoma（OS ） MG-63 cells， and to clarify its possible mechanism. Methods The MG-63 cells were cultured in vitro and divided into control group （without drug intervention） and different concentrations of paeonol groups （given 10， 20， 40， 80， 160，and 320 mg·L^－1 paeonol， respectively）.The survival rates of the MG-63 cells in various groups were detected by CCK-8 method and the half inhibitory concentration （IC₅₀） value was selected as the drug concentration for subsequent experiment. The MG-63 cells were divided into control group（without drug treatment） and paeonol group（given 215.8 mg·L^－1 paeonal）.The expression levels of CXCR4， interleukin 6 （IL-6），and phosphorylated STAT3 （p-STAT3），and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method.The MG-63 cells were divided into control group， paeonol group， paeonol+empty group （given 215.8 mg·L^－1 paeonol+empty plasmid）， and paeonol+overexpression group （given 215.8 mg·L^－1 paeonol+CXCR4 over-expression plasmid）. The scratch healing rates of the MG-63 cells in various groups were detected by cell scratch experiment， the clone formation rates the MG-63 cells in various groups were detected by cloning formation experiment， and the expression levels of CXCR4， IL-6， p-STAT3， and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method. Results Compared with control group， the survival rates of the MG-63 cells treated in 40， 80， 160， and 320 mg·L^－1 paeonol groups were decreased （P<0.05）， and the IC₅₀ value of paeonol was 215.8 mg·L^－1. Compared with control group， the expression levels of CXCR4， IL-6， and p-STAT3 proteins in the MG-63 cells in paeonol group were decreased （P<0.05）. Compared with control group， the scratch healing rates and clone formation rates of the MG-63 cells in paeonol group and paeonol+empty group were decreased （P<0.05）， and the expression levels of CXCR4， IL-6，and p-STAT3 proteins in the MG-63 cells were decreased （P<0.05）； compared with paeonol group，the scratch healing rate， clone formation rate， expression levels of CXCR4， IL-6， p-STAT3，and STAT3 proteins in the MG-63 cells in paeonol+empty group had no significant differences（P>0.05）； compared with paeonol group， the scratch healing rate and clone formation rate of the MG-63 cells in paeonol+overexpression group were increased （P<0.05）， and the expression levels of CXCR4， IL-6 and p-STAT3 proteins were increased （P<0.05）. Conclusion Paeonol can inhibit the proliferation， migration，and clonogenic ability of the MG-63 cells， and its mechanism may be related to the regulation of CXCR4/STAT3 signal pathway.

Objective To discuss the effect of microRNA-491-5p（miR-491-5p） over-expression on the proliferation and migration of the human nasopharyngeal carcinoma HONE-1 cells， and to provide the evidence for the study of the pathogenesis and targeted therapy of nasopharyngeal carcinoma. Methods The human nasopharyngeal carcinoma HONE-1 cells were divided into control group （transfected with control plasmid） and miR-491-5p group （transfected with miR-491-5p plasmid）. The transfection efficiencies of the HONE-1 cells in two groups were observed under fluorescence microscope； the proliferation activities of the HONE-1 cells in two groups were detected by CCK-8 assay； the number of migration HONE-1 cells in two groups were detected by Transwell chamber assay；the expression levels of Vimentin and E-cadherin mRNA in the HONE-1 cells in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR）method； the expression levels of Vimentin and E-cadherin proteins in the HONE-1 cells in two groups were detected by Western blotting method. Results Compared with control group， the proliferation activities（t=2.832， P=0.0473； t=4.522， P=0.0014； t=9.308， P<0.01） and number of migration HONE-1 cells （t=9.639， P<0.01） in miR-491-5p group at 24，48，and 72 h after treatment were significantly decreased； compared with control group， the expression levels of Vimentin mRNA （t=7.535， P=0.0017） and protein （t=7.219， P=0.0187） in the HONE-1 cells in miR-491-5p group were significantly decreased， and the expression levels of E-cadherin mRNA （t=4.88， P=0.0395） and protein （t=5.754， P=0.0289） were significantly increased. Conclusion Over-expression of miR-491-5p can inhibit the proliferation， migration， and epithelial-mesenchymal transition （EMT） of the human nasopharyngeal carcinoma cells.

Objective To observe the changes of sagittal plane balances of cervical spine of the patients underwent posterior cervical single open-door expansive laminoplasty， and to provide the imaging evidence for the postoperative rehabilitation training of the patients. Methods A total of 32 patients who underwent posterior cervical single open-door expansive laminoplasty were selected. According to the median value（15.75 mm ）of sagittal vertical axis （SVA） distance before operation， the patients were divided into low SVA group and high SVA group，and there were 16 patients in each group. The imaging and clinical data of the patients in two groups before operation and at the last follow-up were retrospectively analyzed. The SVA value， cervical lordosis angle （Cobb angle）， and T1 tilt angle （T1s） of the patients at the X-ray lateral view of the cervical spine were detected before operation and at the last follow-up. The postoperative Japanese Orthopaedic Association （JOA） score， neck disability index （NDI） store，and satisfaction score of the patients in two groups were analyzed. Results Compared with before operation， the NDI score of the patients in high SVA group after operation was decreased （P<0.01）， the JOA score was increased （P<0.01）. Compared with before operation， the NDI score of the patients in low SVA group was decreased （P<0.01）， the JOA score was increased （P<0.01）， and the SVA value was increased （P<0.01）， while there were no significant differences in the Cobb angle and T1s （P>0.05）. There was no statistically significant difference in the occurrence of axial symptoms of the patients between low SVA group and high SVA group（P>0.05）. Conclusion In the follow-up of at least 2 years after operation， posterior cervical single open-door expansive laminoplasty has the certain effect on the sagittal plane balance of cervical spine of the patients. The main manifestations are tendency of cervical lordosis and anterior shift of the center of gravity， but the overall stability can still be maintained. The patients with high SVA have a higher incidence of axial symptoms after operation.

Objective：To discuss the application of the modified needle-through-needle puncture technique in unilateral total hip arthroplasty in the elderly patients，and to clarify its anesthetic effect and feasibility in the single isospecific gravity spinal anesthesia guided by ultrasound. Methods Sixty elderly patients underwent elective unilateral hip arthroplasty with single isobaric gravity spinal anesthesia were selected. The puncture was performed in the patients with the affected side upper in the lateral position. Sixty patients were divided into tradition group and modification group， and there were 30 patients in each group. After the ultrasonic localization of spinous process space， the single subarachnoid puncture through median approach was performed in all the patients， and the successful puncture was determined by the outflow or drawn-out of cerebrospinal fluid in the spinal anesthesia needle.The patients in tradition group were treated with the traditional needle-through-needle puncture technique （a 16 G epidural puncture needle was used to guide the puncture，and a 25 G pencil-point side-hole spinal needle was applied）；the patients in modification group were treated with the modified needle-through-needle puncture technique （a 20 G injection needle was used to break the skin directly for guidance， and a 25 G pencil-point side-hole spinal needle was applied）. The once-puncture success rates，twice-puncture success rates，first-skin-attempt puncture success rates， and twice-skin-attempt puncture success rates， puncture time， patients’ satisfaction rates，onset-time of anesthesia，assessment of anesthesia block， fixation-time of anesthesia， satisfaction of surgeon for muscle relaxation， operation time， residence time in PACU， immediate adverse reactions during anesthesia， and adverse reactions within 48 h after operation of the patients in two groups were compared. Results All the patients had successful punctures. Compared with tradition group， the once-puncture success rate， twice-puncture success rate， first-skin-attempt puncture success rate， and twice-skin-attempt success rate of the patients in modification group were increased （P<0.01），the puncture time was shorter （P<0.01）， the patients’ satisfaction rate was increased （P<0.01）， the onset-time of anesthesia was shortened（P<0.01），the fixation-time of anesthesia block was shortened（P<0.01）， the satisfaction rate of the surgeon was increased （P<0.01），the mean arterial pressure（MAP） and heart rater（HR） of the patients 10 min after anesthsia and 10 min after operation were increased（P<0.05）.Compared with tradiation group， the incidence of immediate adverse reactions during anesthesia， and the incidence of adverse reactions within 48 h after operation of the patients in modification group had no significant differences （P>0.05）. Conclusion In the elderly patients underwent unilateral total hip arthroplasty， the single isospecific gravity spinal anesthesia under non-visual conditions by the modified needle-through-needle puncture technique can significantly shorten the number of punctures and puncture time，and has higher puncture success rate.

Objective To construct a pig-mouse liver cell chimeric model，and to discuss the relevant conditions for the transplantation and regeneration of the porcine hepatocytes without T lymphocyte， B lymphocyte， and NK lymphocyte mediated rejection， and to evaluate the settlement， proliferation， and secretion of functional factors of the porcine hepatocytes in xenograft receptors， and to provide the basis for improving the reconstruction efficiency of the porcine hepatocytes in pig derived mouse models. Methods The porcine hepatocytes were transplanted into the Fah^－/－Rag2^－/－ IL2Rg ^－/－ （FRG） mice by using intrasplenic injection method to construct the transplantation model. Sixteen FRG mice were randomly divided into control group， blank control group， experimental group， and green fluorescence protein（GFP） gene experimental group， and there were four mice in each group.The administration of nitisinone 2-2-nitro-4-（trifluoromethyl）cyclohexane-1，3-dione（NTBC） of the mice in blank control group was stopped，and the body weight was monitored； the mice in control group were injected with the porcine hepatocytes， the administration of NTBC was stopped one week before transplantation， and the body weight was monitored after transplantation. When the body weight was decreased by 20%， the NTBC was administered for 3 d. The mice in experimental group were injected with the porcine hepatocytes， and the administration of NTBC was stopped one week before transplantation； the body weight was monitored after transplantation， and when the body weight was decreased by 20%， the NTBC was administered for 3 d. The mice in GFP gene experimental group were injected with the porcine hepatocytes carrying the GFP gene， the administration of NTBC was stopped one week before transplantation， and the body weight was monitored after transplantation； when the body weight was decreased by 20%， the NTBC was administered for 3 d. The body weights and survival status of the mice in various groups were observed. The levels of alanine aminotransferase （ALT） in serum of the mice in various groups were detected；the expressions of CD19+， CD3+， and NK1.1+cells in peripheral blood of the mice in various groups were detected by flow cytometry；the expression levels of porcine albumin in serum of the mice in various groups were detected by ELISA assay； the regeneration efficiencies of the porcine hepatocytes of the mice in various groups were detected by immunohistochemistry. Results Compared with method of collagenase digestion after shearing， the digestion process of the two-step perfusion method was milder and had less damage to cells， and the cell activity achieved 92%. After stopping the administration of NTBC， the body weight of the mice in control group was gradually decreased and the vitality was gradually decreased. Death began to occur in the mice on the 28th day after continuous stopping the administration.The irreversible cellular damage in liver tissue of the mice could be seen such as liver cell enlargement， cytoplasmic looseness and transparency， and nuclear lysis. The ALT levels in serum of the mice in blank control group was gradually increased after stopping the administration. After re-administration of NTBC， the body weight of the mice in control group was gradually returned to the normal levels. The flow cytometry results showed that the CD19+， CD3+， and NK1.1+cells in peripheral blood of the mice could not be seen after stopping the administration of NTBC. The immunohistochemistry staining results showed that no dark Fah+positive cells were found in liver tissue of the mice in experimental group， while the dark Fah+positive cells were observed in liver tissue of the transplanted mice； the regeneration efficiency of porcine hepatocytes was （11 ± 4）%. The regeneration efficiency of porcine hepatocytes of the mice in GFP gene experimental group was （10 ± 2）% when stimulated by three color lasers in the same field of view. Conclusion The pig derived FRG mouse liver chimeric models are successfully constructed， and the mouse models for long-term stable expansion of the porcine hepatocytes are established.

Objective： To analyze the serum metabolite compositions of the patients with esophageal squamous cell carcinoma （ESCC）， to screen out the potential differential metabolites， and to explore the relationship between the occurrence of ESCC and energy metabolism， and to provide the useful energy metabolites for the early diagnosis of ESCC. Methods The serum samples of the ESCC patients and healthy volunteers were collected and divided into ESCC group and control group， and there were 7 cases in each group. Liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was used to detect the serum metabolites of the subjects in two groups；partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） models were used to identify the differences in serum metabolites of the subjects between ESCC group and control group， and they were classified according to the chemical classification of serum metabolites； targeted metabolomics analysis on serum energy metabolites was conducted based on the multi-response monitoring （MRM） mode to further identify the significantly differential energy metabolites and their pathways. Results A total of 266 metabolites were identified in the serum samples of the subjects in ESCC group and control group with the positive and negative ion modes， and there were 164 and 102 metabolites identified with the above two modes， respectively. The univariate statistical analysis results showed that there were significant differences in serum metabolites of the subjects between ESCC group and control group. Among the metabolites with chemical classification， lipids and lipid-like molecules accounted for the highest proportion （18.421%）. Three significantly different energy metabolites in ESCC group were found， and the expression levels of L-malic acid and isocitric acid were increased and the expression level of cyclic adenosine monophosphate （cAMP） was decreased. Conclusion There are differences in the expressions of serum energy metabolites between the ESCC patients and healthy controls， and the occurrence of ESCC is related to the energy metabolism.

Objective To discuss the inhibitory effect of bone marrow-derived mesenchymal stem cells （BMSCs） over-expressing forkhead box transcription factor 1 （FOXO1） on the eosinophil infiltration and airway remodeling of the asthmatic mice， and to clarify the possible mechanism. Methods The BMSCs of the mice were isolated， and Oil Red O staining and Alizarin staining were used to identify the BMSCs. Flow cytometry was used to identify the phenotypes of the BMSCs. The BMSCs at logarithmic growth phase were collected and divided into control group （without treatment）， FOXO1-BMSCs group （infected with recombinant lentivirus carrying FOXO1 gene）， and NC-BMSCs group （infected with negative control recombinant lentivirus）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of FOXO1 mRNA in the BMSCs in various groups； Western blotting method was used to detect the expression levels of FOXO1 protein in the BMSCs in various gorups. Forty mice were randomly divided into control group （given saline）， model group （given saline）， NC-BMSCs group （given NC-BMSCs cell suspension）， and FOXO1-BMSCs group （given FOXO1-BMSCs cell suspension），and there were 10 mice in each group. Except for control group， the mice in the other three groups were sensitized with ovalbumin （OVA） and challenged with aerosol to establish the asthma models. The cell classification counts in bronchoalveolar lavage fluid （BALF） of the mice in various groups were detected；the histopathology of lung tissue of the mice in various groups was observed by HE staining；the expressions of α-smooth muscle actin （α-SMA） and proliferating cell nuclear antigen （PCNA） in lung tissue of the mice in various groups were detected by immunofluorescence method； the bronchial lumen circumference （Pi）， wall area （W）， smooth muscle area （S）， and numbers of smooth muscle cell nuclei （N） of the mice in various groups were detected by using Image-Pro Plus Software，and the ratios of S/Pi， W/Pi， and N/Pi were calculated； the expression levels of matrix metalloproteinase-9 （MMP-9），matrix metalloproteinase-12 （MMP-12）， and tissue inhibitor of metalloproteinase-1 （TIMP-1） proteins in lung tissue of the mice in various groups were detected by Western blotting method. Results The isolated and cultured BMSCs showed a spindle-shaped morphology， and significant red lipid droplets and orange-red precipitates could be observed after Oil Red O and Alizarin staining. The expression amonuts of CD29， CD44， and CD71 on surface of the BMSCs were increased， while the expression amonuts of CD34， CD45， and HLA-DR were decreased. Compared with control group and NC-BMSCs group， the expression levels of FOXO1 mRNA and protein in the BMSCs in FOXO1-BMSCs group were increased （P<0.05）. Compared with model group and NC-BMSCs group， the counts of eosinophils， macrophages， lymphocytes， and neutrophils in bronchoalveolar lavage fluid （BALF） of the mice in FOXO1-BMSCs group were decreased（P<0.05）， and the inflammatory cell infiltration in the airways and alveoli was alleviated； the ratios of S/Pi， W/Pi， and N/Pi were decreased（P<0.05）， and the expression amounts of α-SMA and PCNA in lung tissue were decreased， the expression levels of MMP-9， MMP-12， and TIMP-1 proteins in lung tissue were decreased （P<0.05）. Conclusion The BMSCs over-expressing FOXO1 can attenuate the inflammatory cell infiltration， particularly eosinophil infiltration， and inhibit airway remodeling in the asthmatic mice， thus alleviating the asthma symptoms.

Objective To analyze the key genes and candidate pathways related to the occurrence and development of glioblastoma multiforme （GBM） by bioinformatics methods， and to explore the pathogenesis and therapeutic targets of GBM. Methods Gene Expression Datasets TCGA-GBM and GSE7696 were obtained from The Cancer Genome Atlas （TCGA） Database and Gene Expression Omnibus （GEO） Database. Deseq2 and limma R Data Packages were used to screen the differentially expressed genes （DEGs） in GBM tissue and adjacent normal tissue， and the Gene Ontology （GO） fuctional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed on the DEGs； the protein-protein interaction （PPI） network was analyzed by using the STRING Database； Cytoscape 3.9.1 Software was used to visualize the PPI network and perform the modular analysis. Results After the DEGs analysis of TCGA-GBM Transcription Data and Chip Dataset GSE7696， a total of 13 upregulated differentially expressed genes （UDEGs） and 77 downregulated differentially expressed genes （DDEGs） were obtained. The results of GO fuctional enrichment analysis showed that the DDEGs were mainly concentrated in the chloride channel activity， gamma-aminobutyric acid （GABA） receptor activity， GABA-gated chloride ion channel activity， GABA-A receptor activity， anterograde trans-synaptic signaling， chemical synaptic transmission and other biological processes. The KEGG signaling pathway were mainly concentrated in the GABA ergic synapse， neuroactive ligand-receptor interaction， neuro synapses containing serum， synaptic vesicle cycle and other signaling pathways. Two important gene modules were identified by PPI and module construction. The Cytoscape Software analysis results showed that solute carrier family 17 member 6（SLC17A6）， solute carrier family 1 member 2（SLC1A2），tachykinin precursor 1（TAC1），synaptotagmin 1（SYT1）， RNA binding fox-1 homolog 3（RBFOX3）， and gamma-aminobutyric acid type A receptor subunit gamma 2（GABRG2） were the key genes in PPI network. Conclusion SLC17A6，SLC1A2，TAC1，SYT1，RBFOX3，and GABRG2 genes may be involved in the occurrence and development of GBM，and the dysregulation of GABA ergic synaptic transmission related genes and pathway regulation network may be the main mechanism of pathogenesis of GBM.

Objective To discuss the effect of macrophage-derived extracellular vesicle long non-code RNA（lncRNA） highly up-regulated in liver cancer（HULC） on the metastasis of hepatocellular carcinoma （HCC），and to elucidate its mechanism. Methods The bone marrow mononuclear cells were isolated from the mice and differentiated into bone marrow-derived macrophages （BMDM） through phorbol myristate acetate induction. Interleukin-4 （IL-4） was used to induce the M2 macrophage polarization，regarded as tumor-associated macrophages（TAM）.The TAM-derived extracellular vesicles （referred to as TAM-exos） were collected using differential centrifugation. RNA interference was used to transfect TAM with lncRNA HULC siRNA or negative control （NC） plasmids， and the extracellular vesicles secreted by TAM in various groups were extracted （referred to as lncRNA HULC-siRNA-exos and NC-exos）. The HepG2 cells were divided into control group and TAM-exos group； NC-exos group and lncRNA HULC-siRNA-exos group； lncRNA HULC-siRNA-exos group and lncRNA HULC-siRNA-exos+SKL2001 group according to the purpose of the experiment. After the corresponding treatments， Transwell champer assay was used to detect the numbers of migration and invasion HepG2 cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of lncRNA HULC in the HepG2 cells in various groups； Western blotting method was used to detect the expression levels of Wnt3A， β-catenin， c-Myc， and Cyclin D1 in the HepG2 cells in various groups. The nude mouse orthotopic liver cancer transplantation model was established， and the mice were divided into control group， NC-exos group， TAM-exos group， and lncRNA HULC-siRNA-exos group. The numbers of lung metastatic lesions in the nude mice in various groups after treatment with M2 macrophage-derived extracellular vesicles containing lncRNA HULC were detected. Results The TAM-exos displayed the morphology and biological characteristics of extracellular vesicles. Compared with control group， the numbers of migration and invasion HepG2 cells in TAM-exos group were increased（P<0.05）， and the expression levels of lncRNA HULC， Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells in TAM-exos group were increased （P<0.05）. Compared with NC-exos group， the numbers of migration and invasion HepG2 cells in lncRNA HULC-siRNA-exos group were decreased，and the expression levels of lncRNA HULC， Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells were decreased （P<0.05）. Compared with lncRNA HULC-siRNA-exos group， the numbers of migration and invasion HepG2 cells in lncRNA HULC-siRNA-exos+SKL2001 group were increased（P<0.05），and the expression levels of Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells were increased （P<0.05）. The nude mouse experiment results showed that compared with control group， the number of lung metastatic lesions of the nude mice in TAM-exos group was increased （P<0.05）； compared with TAM-exos group and the NC-exos group， the number of lung metastatic lesions of the nude mice in lncRNA HULC-siRNA-exos group was decreased （P<0.05）. Conclusion The TAM-derived extracellular vesicle lncRNA HULC promotes the invasion and metastasis of the HCC cells， and its mechanism may be associated with the activation of the Wnt signaling pathway.

Objective To discuss the effect of exogenous CXC chemokine ligand 10（CXCL10） on the proliferation and migration of the hepatocellular carcinoma（HCC）SMMC-7721 cells， and to clarify its mechanism. Methods The human HCC SMMC-7721 cells were divided into 0 mg·L^－1 CXCL10 group，10 mg·L^－1 CXCL10 group，and 30 mg·L^－1 CXCL10 group according to the CXCL10 concentration. Some of the above cells were treated with extracellular regulated protein kinase（ERK） inhibitor PD98059 （80 μmol·L^－1）， then the SMMC-7721 cells were divided into 0 mg·L^－1 CXCL10+PD98059 group， 10 mg·L^－1 CXCL10+PD98059 group， and 30 mg·L^－1 CXCL10+PD98059 group. The proliferation rates of the SMMC-7721 cells in various groups were detected by CCK-8 method； the EdU positive expression rates in SMMC-7721 cells in various groups were detected by EdU method； the migration rates of the SMMC-7721 cells in various groups were detected by Transwell chamber assay； the expression levels of ERK， phosphorylated ERK（p-ERK）， and Cyclin D1 proteins in the SMMC-7721 cells in various groups were detected by Western blotting method. Results The CCK-8 results showed that after cultured for 24 h， compared with 0 mg·L^－1 CXCL10 group， the proliferation rates of the cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.05 or P<0.01）；the EdU detection results showed that compared with 0 mg·L^－1CXCL10 group， the positive expression rates of EdU in the cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1CXCL10 group were increased （P<0.01）. The Transwell chamber assay results showed that after cultured for 48 h， compared with 0 mg·L^－1 CXCL10 group， the migration rates of the cells in 10 mg·L^－1CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.01）.The Western blotting results showed that after cultured for 24 h and treated with CXCL10 for 24 h，compared with 0 mg·L^－1 CXCL10 group， the expression levels of ERK， p-ERK，and Cyclin D1 proteins in the SMMC-7721 cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.05）. The CCK-8 method results showed that after treated with ERK inhibitor PD98059， compared with 0 mg·L^－1 CXCL10 group， the proliferation rates of the SMMC-7721 cells in 10 mg·L^－1 CXCL10+PD98059 group and 30 mg·L^－1 CXCL10+PD98059 group were decreased （P<0.05）； compared with 10 mg·L^－1 CXCL10 group， the proliferation rate of the SMMC-7721 cells in 10 mg·L^－1 CXCL10+PD98059 group was decreased （P<0.05）； compared with 30 mg·L^－1 CXCL10 group， the proliferation rate of the SMMC-7721 cells in 30 mg·L^－1 CXCL10+PD98059 group was decreased （P<0.05）. Conclusion CXCL10 can promote the proliferation and migration of the HCC SMMC-7721 cells， and its mechanism is mainly related to the up-regulation of the expressions of ERK/p-ERK/Cyclin D1 pathway proteins.

Objective To identify the key genes associated with the early diagnosis and poor prognosis of hepatitis B virus-associated hepatocellular carcinoma （HBV-HCC） by using the bioinformatics methods， and to elucidate the underlying molecular mechanism of occurence and development of HBV-HCC. Methods Gene Expression Omnibus （GEO） Database was used to retrieval “hepatitis B induced HCC”； the Gene Dataset GSE121248 was downloaded， the differentially expressed genes （DEGs） were screened by the “limma” Data Package in R Software， and the DEGs were enriched by using the “clusterProfiler” Data Package for Gene Ontology （GO） functional analysis and the Kyoto Genes and Genome Encyclopedia （KEGG） signaling pathway enrichment analysis； STRING Database and Cytoscape Software were used to establish the protein-protein interaction（PPI） network and the key genes were screened out.Gene Expression Profiling Interactive Analysis（GEPIA）， Kaplan Meier-Plotter，and Human Protein Atlas（HPA）Databases were used to verify the key genes and the expression levels of proteins；the infiltration of the immune cells was analyzed based on the “CIBERSORT” Data Package. Results A total of 574 DEGs were identified，including 173 up-regulated genes and 401 down-regulated genes. The GO functional enrichment analysis results showed that DEGs were mainly enriched in the biological processes such as small molecule metabolism， signal transduction， immune response， inflammatory response，and so on； the KEGG signaling pathway enrichment analysis results showed that the DEGs were mainly enriched in retinol metabolism， cytochrome P450 metabolic pathway of exogenous drugs， and chemical carcinogenesis，and so on. The PPI network results showed that cell division cycle 20（CDC20），cyclin dependent kinase 1（CDK1）， cyclin A2（CCNA2）， pindle checkpoint protein（BUB1B）， topoisomerase Ⅱ α（TOP2A）， discs large homolog associated related protein 5（DLGAP5）， abnormal spindle-like microcephaly associated protein（ASPM）， centrosomal protein 55（CEP55）， kinesin superfamily 11（KIF11），and kinesin superfamily 20A（KIF20A） were the key genes. The GEPIA Database analysis results showed that these above 10 key genes were highly expressed in the HCC patients. The Kaplan Meier survival curve showed that the overall survivals of the HCC patients with high expression of key genes were shorter than those of the HCC patients with low expression of key genes. Conclusion The genes related to cell cycle and viral oncogenesis （CDC20， CDK1， CCNA2， and BUB1B） are closely associated with the occurence and development and poor prognosis of the HBV-HCC patients， which may become the diagnostic markers and new targets for the treatment.

Objective To discuss the effect of apolipoprotein C1 （APOC1） expression on the proliferation and apoptosis of the hepatocellular carcinoma cells， and to preliminarily clarify the related molecular mechanism. Methods The expression level of APOC1 mRNA in hepatocellular carcinoma tissue and its relationship with the prognosis of the patient were analyzed by The Cancer Genome Atlas （TCGA） Database； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of APOC1 mRNA in different hepatocellular carcinoma cells； the human liver cancer HepG2 cells with low APOC1 expression were selected as the subjects. The HepG2 cells were transfected with pcDNA3.1-APOC1 plasmid to over-express APOC1 （APOC1 over-expression group），and the HepG2 cells transfected with empty vector pcDNA3.1 were regarded as control group. MTS assay and 5-ethynyl-2'-deoxyuridine （EdU） staining were used to detect the proliferative activities and proliferation rates of the cells in two groups； Transwell chamber assay was used to detect the numbers of migration cells in two groups；flow cytometry and TUNEL assay were used to detect the percentages of the cells at different cell cycles and apoptotic rates in two groups；Western blotting method was used to detect the expression levels of extracellular regulated protein kinase （ERK）， phosphorylated ERK （p-ERK）， protein kinase B （AKT）， phosphorylated AKT （p-AKT）， B-cell lymphoma-2 （Bcl-2）， and cleaved cysteinyl aspartate specific proteinase-3（cleaved caspase-3） proteins in the cells in two groups. Results The TCGA Database results showed that the expression level of APOC1 mRNA in hepatocellular carcinoma tissue was lower than that in normal liver tissue （P<0.05）， and the patients with low expression of APOC1 mRNA had poor prognosis. The RT-qPCR results showed that the expression level of APOC1 mRNA in the HepG2 cells was the lowest， and the HepG2 cells were chosen for the subsequent research. Compared with control group， the proliferative activity and proliferation rate of the cells in APOC1 over-expression group were decreased （P<0.05 or P<0.01），the number of migration cells was decreased （P<0.01），and the percentage of the cells at S phase and the apoptotic rate were significantly increased （P<0.01）. Compared with control group， the expression levels of p-ERK， p-AKT， and Bcl-2 proteins in the cells in APOC1 over-expression group were significantly decreased （P<0.05），and the expression level of cleaved caspase-3 protein was increased （P<0.01）. Conclusion High expression of APOC1 can inhibit the proliferation of the human liver cancer HepG2 cells and induce the apoptosis，and its mechanism may be related to inhibition of the expressions of p-ERK，p-AKT，Bcl-2 proteins and promotion of the expression of cleaved caspase-3 protein.

Objective To discuss the effect of downregulating the proline-rich protein 11 （PRR11） expression on drug resistance of the esophageal cancer drug resistant cells， and to clarify the related mechanism. Methods The drug resistant cells EC9706/cisplatin（DDP） were established by incrementally stimulating the human esophageal cancer EC9706 cells with the increasing concentrations of DDP. The drug sensitivity of the EC9706/DDP cells was detected by MTT assay； the expression levels of PRR11 mRNA and protein in the EC9706/DDP cells and their parent EC9706 cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. The EC9706/DDP cells were divided into control group，sh-NC group （infected with sh-NC），sh-PRR11 group（infected with sh-PRR11）， sh-NC+DDP group （infected with sh-NC and treated with 4 mg·L^－1 DDP）， and sh-PRR11+DDP group （infected with sh-PRR11 and treated with 4 mg·L^－1 DDP）. The expression levels of PRR11 mRNA in the cells in various groups were detected by RT-qPCR method； the expression levels of PRR11， phosphoinositide 3-kinase （PI3K） p110α， protein kinase B （AKT）， phosphorylated AKT （p-AKT）， P-glycoprotein （P-gp）， and multidrug resistance-associated protein 1 （MRP1） proteins in the cells in various groups were detected by Western blotting method；the apoptotic rates of the cells in various groups were detected by flow cytometry. Results The DDP-resistant cell line EC9706/DDP was successfully obtained，and the drug resistance index was 7.23±0.86. Compared with the EC9706 cells， the expression levels of PRR11 mRNA and protein in the EC9706/DDP cells were increased （P<0.05）. Compared with control and sh-NC groups， the expression levels of PRR11 mRNA and protein in the cells in sh-PRR11 group were decreased （P<0.05），and the 50% inhibitory concentration （IC₅₀） of DDP was decreased （P<0.05）.Compared with sh-NC group，the expression levels of PI3K p110α， p-AKT， P-gp， and MRP1 proteins in the cells in sh-NC+DDP and sh-PRR11 groups were decreased （P<0.05），and the apoptotic rate of the cells was increased （P<0.05）. Compared with sh-NC+DDP group and sh-PRR11 group，the expression levels of PI3K p110α， p-AKT， P-gp， and MRP1 proteins in the cells in sh-PRR11+DDP group were increased（P<0.05），and the apoptotic rate of the cells was increased（P<0.05）. Conclusion Downregulating the expression of PRR11 gene in the drug resistant EC9706/DDP cells can inhibit the expressions of drug resistance-related proteins， reverse the resistance to DDP， and induce the apoptosis； its mechanism may be related to the inhibition of activation of the PI3K/AKT signaling pathway.

Objective To discuss the effect of downregulating of high mobility group box protein 2 （HMGB2） expression on the biological behavior of the liver cancer cells and the epithelial-mesenchymal transition （EMT） process， and to clarify its mechanism. Methods The human liver cancer LM3 cells at logarithmic growth phase were divided into negative control group and HMGB2 RNA interference group （HMGB2 siRNA group）；the cells in two groups were transfected with RNA oligonucleotides（RNA oligos） with irrelevant sequences and RNA oligos designed to knock down HMGB2， and the Lipofectamine 2000 was regarded as the vector.The expression levels of HMGB2 mRNA and protein in the cells in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods； cell scratch assay and Transwell chamber assay were used to detect the migration and invasion abilities of the cells in two groups； the expression levels of E-cadherin， N-cadherin， and Vimentin proteins and protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway related proteins in the cells in two groups were detected by Western blotting method. Results Compared with negative control group， the expression levels of HMGB2 mRNA and protein in the cells in HMGB2 siRNA group were significantly decreased （P<0.05），the cell scratch healing rate was significantly decreased （P<0.01），the number of invasion cells was significantly decreased （P<0.01），and the expression level of E-cadherin protein in the cells was significantly increased （P<0.01）， while the expression levels of N-cadherin， Vimentin， mTOR， AKT， and phosphorylated AKT （p-AKT） proteins in the cells were significantly decreased （P<0.05 or P<0.01）. Conclusion Downregulating the expression of HMGB2 can reduce the migration and invasion abilities of the liver cancer LM3 cells and inhibit the EMT，and its mechanism may be related to regulating the expression of the AKT/mTOR pathway related proteins.

Early start denver model (ESDM) is a comprehensive early intervention approach for the children with autism spectrum disorder (ASD) between 12-month-old－36-month-old. The model is built upon the theoretical foundations of applied behavior analysis, denver model (DM), and pivotal response treatment, and it is one of the naturalistic developmental behavioral interventions. Compared with the other early intervention methods, ESDM is not limited by the environment of intervention; it encompasses all the areas of development during teaching practice and has been widely adopted for the early intervention of the children with ASD, and achieves the satisfactory therapeutic effect. The ESDM typically uses an intensive one-on-one intervention approach, but variabilities have emerged in its practical application, such as group ESDM(G-ESDM), parent-implemented ESDM (P-ESDM), and peer-mediated ESDM. In particular,G-ESDM and P-ESDM have provided the learning opportunities for more families, showing a broad application prospect. This study reviews the theoretical foundations, teaching models, and the effects of various intervention modalities of the ESDM in the treatment of ASD; combined with the domestic and international research findings,this study offers a reference for further studies on the mechanism of ESDM intervention for ASD.

Objective To discuss the expression of programmed cell death-ligand 1 （PD-L1） in the oral squamous cell carcinoma （OSCC） cells and its effect on biological behavior of the OSCC CAL27 cells， and to clarify the possible mechanism. Methods Western blotting method was used to detect the expression levels of PD-L1 protein in the oral epithelial HOK cells and OSCC CAL27， TCA8113， and SCC15 cells； immunofluorescence staining method was used to detect the expression and localization of PD-L1 protein in the CAL27 cells. The CAL27 cells were divided into control group （transfected with si-NC） and si-PD-L1 group （transfected with si-PD-L1）. Western blotting method was used to detect the interference efficiency of the cells in two groups； CCK-8 assay was used to detect the proliferative activities of the cells in two groups at different time points； plate clone formation assay was used to detect the numbers of clone formation of the cells in two groups； cell scratch healing assay was used to detect the scratch healing rates of the cells in two groups； Transwell chamber assay was used to detect the numbers of migration and invasion cells in two groups. Results The expression level of PD-L1 protein in the OSCC cells was higher than that in the HOK cells （P<0.05 or P<0.01）； PD-L1 expressed in the cytoplasm and nucleus of the CAL27 cells. The CCK-8 assay and plate clone formation assay results showed that compared with control group， the proliferative activities of the CAL27 cells in si-PD-L1 group at different time points were significantly decreased（P<0.05 or P<0.01）， and the numbers of clone formation were significantly decreased （P<0.01）. The cell scratch healing assay results showed that compared with control group， the scratch healing rates of the cells in si-PD-L1 group were significantly decreased （P<0.05 or P<0.01）. The Transwell chamber assay results showed that compared with control group， the numbers of migration and invasion cells in si-PD-L1 group were significantly decreased （P<0.01）. Conclusion The expression of PD-L1 in the OSCC cells is higher than that in normal oral epithelial cells， and knocking down PD-L1 expression can inhibit the proliferation， clone formation， migration and invasion capabilities of the OSCC cells.

Objective To select the differential prognostic lactic acid metabolism-related genes （LRGs） of the head and neck squamous cell carcinoma （HNSCC） to construct the LRGs prognostic model of HNSCC， and to clarify the potential mechanism. Methods The HNSCC gene expression and clinical data were obtained from The Cancer Genome Atlas （TCGA） and Gene Expression Omnibus （GEO） Databases， the LRGs were identified through GeneCards Database， and R software was used to screen out the LRGs of HNSCC； univariate Cox regression analysis was used to identify prognosis-related genes； two different subtypes were identified based on the prognostis-related LRGs； Kaplan-Meier （K-M） curve analysis was used to compare the prognosis of the patients between two groups； CIBERSORT algorithm was used to perform the immuno-correlation analysis between two groups；multivariate Cox regression analysis and LASSO regression analysis were used to construct the prognostic model； receiver operating characteristic curve （ROC） and K-M survival curve were used to assess the relationship between LRGs and survival and prognosis of the HNSCC patients. The prognostic model was validated by GSE27020， GSE41613，and GSE65858 datasets.The experiment were grouped based on risk score，and immune-related analysis and tumor score analysis were performed. Results The TCGA Database differential analysis results showed that 1 196 LRGs were identified from HNSCC samples； univariate Cox regression analysis selected 27 differentially expressed genes （DEGs） associated with the prognosis of the HNSCC patients. Two different LRGs subtypes （Group 1 and Group 2） were identified according to the prognosis-related genes. The K-M survival curves results showed that the overall survival （OS） of the patients in Group 2 was significantly higher than that in Group 1， and the immune cell expression amount of the patients in Group 2 was also higher than that in group 1. The multivariate Cox regression and LASSO regression analysis results screened out 9 LRGs， including hypoxanthine phosphoribosyltransferase 1 （HPRT1）， amyloid precursor protein （APP）， glycogen phosphorylase L（PYGL），urokinase-type plasminogen activator（PLAU）， cannabinoid receptor 2 （CNR2）， stanniocalcin 2 （STC2）， nucleotide binding oligomerization domain-like receptor protein 1 （NLRP1）， integrin-linked kinase （ILK）， and forkhead box B1 （FOXB1）；the prognostic model was constructed.The K-M and ROC curve results indicated that the expression levels of above 9 genes were associated with the survival and prognosis of the HNSCC patients， providing good 1-year， 2-year， and 3-year survival prediction effect， and the area under ROC curve （AUC） values were all greater than 0.650. Furthermore， the predictive ability of the prognosis model was validated in GSE27020， GSE41613， and GSE65858 datasets. The patients classified based on the risk scores had distinguishable immune statuses. Conclusion The differentially expressed LRGs of HNSCC screened by bioinformatics methods are related to the survival and prognosis of the HNSCC patients； the prognostic model constructed by 9 LRGs can predict the survival status and treatment response of the HNSCC patients.

Objective To observe the clinical efficacy of plan-do-check-Act （PDCA） cycle management model combined with pulsed tooth punch applied in maintenance period of the patients with moderate to severe periodontitis， and to provide the theoretical basis for application of the PDCA cycle management model in the periodontitis patients. Methods A total of 50 patients with moderate to severe periodontitis were selected based on predefined inclusion， exclusion， and elimination criteria. The patients were randomly divided into experiment group （n=25） and control group （n=25）. The patients in experiment group underwent maintenance care with pulsed tooth punch in combination with the BASS brushing technique， while the patients in control group maintained oral hygiene with the BASS brushing technique alone.The patients in both two groups were managed with the PDCA cycle management model. The patients were asked to return for follow-up visits at 2， 4， 8， and 12 weeks of self-care， and the personalized corrections and guidance were provided based on the plaque accumulation. The clinical periodontal parameters， including plaque index （PLI）， probing depth （PD），and bleeding index （BI），at 4 and 12 weeks of self-care， as well as the levels of tumor necrosis factor-α （TNF-α） and interleukin-17 （IL-17） in the gingival crevicular fluid of the patients in two groups were observed and recorded. Results After 4 and 12 weeks of self-care， compared with control group，the PLI， PD， and BI of the patients in experiment group were significantly decreased （P<0.01）； compared with baseline， the PLI， PD， and BI of the patients in both two groups at 4 and 12 weeks of self-care were increased （P<0.05 or P<0.01）； Compared with 4 weeks of self-care， the PLI， PD， and BI of the patients at 12 weeks of self-care were increased （P<0.01）. After 4 and 12 weeks of self-care， compared with control group，the levels of TNF-α and IL-17 in gingival crevicular fluid of the patients in experiment group were significantly decreased （P<0.01）； compared with baseline，the levels of TNF-α and IL-17 in gingival crevicular fluid of the patients in two groups at 4 and 12 weeks of self-care were increased （P<0.01）. Conclusion The use of pulsed tooth punch under the PDCA cycle management model can significantly decrease the PLI， PD， BI， and the levels of inflammatory factors in gingival crevicular fluid of the patients with moderate to severe periodontitis，and inhibit the plaque formation and control the gingival inflammation， benefite the maintenance of efficacy of the patients with moderate to severe periodontitis.

Objective To screen the interacting protein of ubiquitin-conjugating enzyme E2S （UBE2S） and construct the hepatocellular carcinoma （HCC） based on UBE2S interacting protein prognosis model （UIPM），and to discuss the value of UIPM in assessing the prognosis of the HCC patients. Methods Co-immunoprecipitation （Co-IP） was used to screen the protein complexes binding to Flag-UBE2S. After validation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis （SDS-PAGE） and Western blotting methods；liquid chromatography-mass spectrometer （LC-MS） was used to identify the UBE2S interacting proteins； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were conducted on these proteins； the prognosis-related proteins from The Cancer Genome Atlas （TCGA） were cross-referenced with UBE2S interacting proteins by survival package of R software；the key proteins were extracted through LASSO regression analysis to build the UIPM； the prognostic model risk scoring formula was established. The HCC patients in TCGA were divided into high risk group and low risk group based on median value of the risk scores. The predictive accuracy of UIPM was evaluated by receiver operating characteristic curve （ROC）， and the predictive accuracy was further validated by International Cancer Genome Consortium （ICGC） Database； univariate regression analysis and multivariate Cox regression analysis were used to detect whether the UIPM risk score was an independent prognostic factor for HCC. Furthermore， the nomogram model was built. Results A total of 97 UBE2S interacting proteins were identified through Co-IP combined with LC-MS analysis.The GO functional enrichment analysis and KEGG signaling pathway enrichment analysis results showed that the interacting proteins were closely associated with cysteine-type endopeptidase activity， oxidative stress， and cell death. The TCGA revealed 5 163 HCC prognosis-related proteins； after intersecting with UBE2S interacting proteins， 40 prognosis-related interacting proteins were found. Seven key proteins were determined through LASSO regression analysis， including UBE2S， heat shock protein family A member 8 （HSPA8）， heterogeneous nuclear ribonucleoprotein H1 （HNRNPH1），chaperonin containing TCP1 subunit 3 （CCT3），eukaryotic translation initiation factor 2 subunit 1 （EIF2S1）， receptor for activated C kinase 1 （RACK1）， and actin related protein 2/3 complex subunit 4 （ARPC4）， and the UIPM was constructed. There was significant difference in survival rate of the patients between high risk group and low risk group （P<0.05）. The ROC curve analysis results showed the area under ROC curve（AUC） values of UIPM for predicting 1-year， 2-year， and 3-year survival risk scores of the HCC patients were all greater than 0.7， indicating the model had high predictive accuracy. This was also confirmed by ICGC Database data. The univariate and multivariate Cox regression analysis results showed that the UIPM risk score was an independent prognostic risk factor for the HCC patients （P<0.05）. The nomogram results showed good consistency between predicted survival rate and actual survival rate of the patient. Conclusion A total of 97 interacting proteins that interact with UBE2S may promote the occurence and devolopment of HCC through oxidative stress and dysregulation of ferroptosis pathways. The UIPM risk score is an independent risk factor for the prognosis of HCC and can be used to predict the outcomes of the patients. UBE2S， HSPA8， HNRNPH1， CCT3， EIF2S1， RACK1， and ARPC4 could be regarded as the new biomarkers and therapeutic targets for HCC.

Objective To analyze the improvement effect of monk fruit on diabetic nephropathy（DN） by network pharmacology，and to elucidate its possible related mechanism. Methods The Traditional Chinese Medicine Systems Pharmacology（TCMSP） Database was used to detect the active ingredients and their targets of monk fruit； the DN target genes were screened out by DisGeNET Database and Genecards Database； the key targets of monk fruit against DN were obtained by comparing the monk fruit with DN targets； protein-protein interaction （PPI） network diagram was constructed by STRING Database and Cytoscape software； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed by Cytoscape software.Molecular docking technology was used to predict the binding abilities of the core targets and the main active ingredients of monk fruit. Results The TCMSP Database combined with the selection criteria was used to screen out a total of five active ingredients of monk fruit （ZINC03860434， Perlolyrine， beta-sitosterol， Kaempferol， and Flazin） as well as 85 targets represented by serine/threonine protein kinase 1 （AKT1），transcription factor RELA， c-Jun N-terminal kinase （JUN），and tumor necrosis factor （TNF）. Among them， Kaempferol contained the most targets.Among the 85 targets， 34 were associated with DN.The GO functional enrichment analysis mainly included biological process（BP） such as oxidative stress， regulation of inflammation and apoptosis， and cell signaling transduction.The KEGG enrichment analysis included advanced glycosylation end product（AGE）-receptor of AGE （AGE-RAGE） signaling pathway， TNF signaling pathway， and C-type lectin receptor signaling pathway.The results molecular docking technology of the main active ingredients of monk fruit and DN target proteins showed that 5 kinds of molecular docking engergy were －8.00－－5.00 kJ·mol^－1. Conclusion Kaempferol is the most effective active ingredient in the monk fruit for the treatment of DN， and its mechanism is mainly related to anti-inflammatory.

Objective To discuss the factors related to the prognosis in the alpha fetoprotein （AFP） negative hepatocellular carcinoma （HCC） patients，and to construct the nomogram for predicting the survival time of the patients. Methods The retrospective analysis on data of 2 064 cases of AFP negative HCC patients extracted from the Surveillance， Epidemiology， and End Results （SEER） Database was conducted， and all the patients were divided into training cohort and internal validation cohort at a ratio of 7∶3， and 101 AFP negative HCC patients from the Integrated Traditional Chinese and Western Medicine Hospital in Hunan Province were regarded as the external validation cohort.The univariate Cox regression analysis results were incorporated into the multivariate analysis， and the independent risk factors for the AFP negative HCC patients were obtained by multivariate Cox analysis to build a cancer specific survival （CSS） prognosis nomogram for the AFP negative HCC patients. The predictive efficacy and clinical utility of the nomogram were evaluated by time-dependent receiver operating characteristic curve （ROC）， calibration plots， and decision curve analysis （DCA）. The total score obtained from the nomogram was used for the risk stratification to compare the degree of risk discrimination between the nomogram and the American Joint Committee on Cancer （AJCC） staging system. Results Ten independent risk factors were selected by multivariate Cox regression analysis to construct 3-year， 4-year， and 5-year CSS prognostic nomograms for the AFP negative HCC patients， including the patient’s age， pathological grade， surgical status， radiotherapy status， chemotherapy status， lung metastasis status， tumor size， tumor T stage， tumor M stage， and marital status. The area under curve （AUC） for the 3-year， 4-year， and 5-year time-dependent ROC in the training cohort were 0.807 （95% CI： 0.786－0.828）， 0.804 （95% CI： 0.782－0.826）， and 0.813 （95% CI： 0.790－0.835）， respectively. In the internal validation cohort， they were 0.776 （95% CI： 0.743－0.810）， 0.772 （95% CI： 0.737－0.808）， and 0.789 （95% CI： 0.752－0.826）， and in the external validation cohort， they were 0.773 （95% CI： 0.677－0.868）， 0.746 （95% CI： 0.620－0.872）， and 0.736 （95% CI： 0.577－0.895）. The calibration plots verified that the nomogram fitted well with the perfect line.The DCA curve revealed that the net benefit of the nomogram was significatly higer than that of the AJCC staging system at certain probability thresholds compared with AJCC staging， the nomogram had a better ability to identify high-risk individuals. Conclusion The serum AFP expression is one of the prognostic markers for the HCC patients. For those patients with AFP negative expression in serum， different considerations should be taken. The nomogram model based on multiple risk factors is a promising clinical tool for assessing the CSS in the AFP negative HCC patients.

Objective To discuss the repairment effect of intra-articular injection of adipose derived stem cells （ADSCs） on articular cartilage destruction in the temporomandibular joint osteoarthritis （TMJOA） model rabbits， and to clarify the possible mechanism. Methods Twenty-seven rabbits were randomly divided into control group， model group， and ADSCs group. The ADSCs of the rabbits were extracted and cultured.The rabbit TMJOA model was prepared by monosodium-iodoacetate （MIA） injection technique.The temporomandibular joint cavity of the TMJOA model rabbits in ADSCs group was given two continuous intra-articular injections of 1.0×10⁶ mL^－1 ADSCs， while the rabbits in control and model group were given sequivalent volume of saline into the temporomandibular joint cavity. After 8 weeks， Micro-CT scan was performed on the temporomandibular joints of the rabbits in various groups； the bone volume fraction （BV/TV）， bone surface area/bone volume （BS/BV）， trabecular thickness （Tb.Th）， trabecular separation （Tb.Sp）， and trabecular number（Tb.N） of condyles tissue of the rabbits in various groups were analyzed； HE staining was used to observe the pathomorphology of condyles tissue of the rabbits in various groups； immunohistochemistry was used to detect the localization and expression levels of SRY-related high mobility group box gene 9（SOX9）， matrix metalloproteinase-13 （MMP-13）， and vascular endothelial growth factor （VEGF） proteins in condyles tissue of the rabbits in various groups；Western blotting method was used to detect the expression levels of SOX9， MMP-13， and VEGF proteins in condyles tissue of the rabbits in various groups. Results The micro-CT scan results showed that compared with control group， the BV/TV， Tb.Th， and Tb.N of condyles tissue of the rabbits in model group were significantly decreased （P<0.05）， while the BS/BV and Tb.Sp were significantly increased （P<0.05）； compared with model group， the BV/TV， Tb.Th， and Tb.N in condyles tissue of the rabbits in ADSCs group were significantly increased （P<0.05）， and the BS/BV and Tb.Sp were significantly decreased （P<0.05）. The HE staining results showed that the condylar cartilage surface of the rabbits in control group was smooth with clear layers and intact structure； compared with control group， the surface of condyles tissue of the rabbits in model group was irregular with thickened hypertrophic layer and areas of cell depletion and clustering； compared with model group， the pathological damage of condyles tissue of the rabbits in ADSCs group was significantly decreased.The immunohistochemical staining results showed that compared with control group and ADSCs group， the number of brown granule in condyles tissue of the rabbits in model group was increased， mainly concentrated in the hypertrophic layer，especially in the bone cartilage junction site and the expression levels of SOX9， MMP-13， and VEGF proteins in condyles tissue of the rabbits in model group were significantly increased （P<0.05）； compared with model group， the number of brown granule in condyles tissue of the rabbits in ADSCs group was significantly decreased，and the expression levels of SOX9， MMP-13， and VEGF proteins were significantly decreased（P<0.05）. The Western blotting results showed that compared with control group， the expression levels of SOX9， MMP-13， and VEGF proteins in condyles tissue of the rabbits in model group were significantly increased （P<0.05）； compared with model group， the expression levels of SOX9， MMP-13， and VEGF proteins in condyles tissue of the rabbits in ADSCs group were significantly decreased （P<0.05）. Conclusion Intra-articular injection of ADSCs can effectively repair the cartilage destruction in TMJOA， alleviate the cartilage injury， and mitigate the progression of osteoarthritis.

Objective To conduct the genetic analysis of a family with one patient suffering from juvenile Parkinson’s disease （JP） and discuss the clinical manifestations， genetic mutation characteristics， and treatment plans prompted by PRKN gene compound heterozygous mutations，and to enhance the clinicians’ awareness of this disease. Methods The clinical data of one patient with JP caused by PRKN gene mutations was analyzed， the clinical manifestations and genetic mutation features of the patient were summarized， and the related literatures were reviewed. Results The patient， a 16-year-old male， was admitted to the hospital due to unstable gait， trembling limbs with rigidity in both lower limbs for three years.The examination results revealed a panic gait， clear consciousness， fluent speech， normal muscle strength in limbs， increased “gear-like” muscle tone in both upper limbs， and “lead-pipe” rigidity in both lower limbs；the sensory functions and tendon reflexes were normal. The head， neck， and thoracic magnetic resonance imaging （MRI） results showed no abnormalities. ¹⁸F-fluorodeoxyglucose （¹⁸F-FDG） positron emission tomography/computed tomography （PET/CT）results showed that the head size and shape were normal， the glucose metabolism in the left cerebellum and middle temporal gyrus was slightly decreased， and the glucose metabolism in bilateral thalami， right frontal lobe， parietotemporal lobe， and left medial frontal lobe was increased. The dopamine transporter （DAT） PET/CT results showed that there was no radioactive distribution in the brain cortex and the DAT distribution in the posterior part of both striata was decreased. The whole-exome sequencing results showed the patient had two PRKN gene mutations，such as codons c.8T>A and c.850G>C compound heterozygous mutations，and each mutation was from one parent；the patient’s father carried the c.8T>A mutation， the patient’s mother carried the c.850G>C mutation， and the patient’s sister had the same genetic mutation site as the patient’s father. Conclusion PRKN gene compound heterozygous mutations may be the basis of the disease in this family. Identification of the mutation c.8T>A expands the mutation spectrum of the PRKN gene， and provides the valuable information for the research on the pathogenic genetic mutations of the JP patients.

Polycystic ovary syndrome (PCOS) is a heterogeneous disorder closely associated with reproductive endocrine dysfunction in the women. The etiology and pathogenesis of PCOS remain unclear. PCOS is the result of the combination of endocrine metabolic disorders, genetics, and environmental factors. Hyperandrogenemia (HA) and insulin resistance (IR) are the fundamental pathophysiological changes in the development of PCOS, and their interactions exacerbate the clinical manifestations of the PCOS patients. The family aggregation and twin study results confirm the genetic predisposition of PCOS; the genome-wide association study (GWAS) results confirm some risk loci and candidate genes of PCOS. The unhealthy lifestyle habits and environmental endocrine disruptors also play an important role in the progression of PCOS, and the gut microbita is involved in the pathogenesis of PCOS. This article provides a comprehensively retrospective analysis on the recent studies about PCOS,and reviews both internal factors and external factors related to the etiology and pathogenesis of PCOS.

Objective To analyze the potential therapeutic targets of Huangqin Tang in treatment of colorectal cancer （CRC） by network pharmacology and molecular docking techniques，and to clarify the related molecular mechanism. Methods The active component and target dataset for Huangqin Tang were constructed based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）；the CRC-disease related target dataset was built by Databases such as GeneCards， Online Mendelian Inheritance in Man （OMIM），and pharmacogenetics and Pharmacogenomics Knowledge Base （PharmGKB）. Drug-disease target intersect， Huangqin Tang herbal formula network， and protein-protein interaction （PPI） networks were built by R software， Cytoscape software， and STRING Database； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were conducted by R software and Metascape platform；molecular docking validation was performed with AutoDock and PyMOL software to assess the ligand-receptor binding. Results A total of 136 effective active components of Huangqin Tang were screened， and 242 potential targets were identified for treatment of CRC， including 18 core targets. Five core key targets closely related to CRC， identified through signaling pathway analysis， were protein kinase B1（AKT1）， mitogen-activated protein kinase 3 （MAPK3），proto-oncogene FOS，tumor protein p53 （TP53），and proto-oncogene MYC.The GO functional enrichment analysis results mainly involved various biological processes related to cellular stress responses. The KEGG signaling pathway enrichment analysis results showed that potential targets were highly enriched in the cancer pathway； further analysis on CRC core targets via KEGG signaling pathway revealed involvement primarily in pathways related to endocrine resistance， apoptosis， and epidermal growth factor receptor-tyrosine kinase inhibitor （EGFR-TKI） resistance. The molecular docking results showed that the active components of Huangqin Tang， including quercetin， kaempferol， baicalein， 7-methoxy-2-methyl isoflavone， and naringenin， were stably docked with AKT1， MAPK3， FOS， TP53， and MYC， and quercetin exhibited the best binding with AKT1. Conclusion The active components of Huangqin Tang can treat CRC through multi-target and multi-pathway. The core ligand quercetin and AKT1 may exert the therapeutic effect in CRC by regulating the phosphatidylinositol 3-kinase （PI3K）/AKT and mammalian target of rapamycin （mTOR） signaling pathways to influence the cell proliferation， differentiation， and apoptosis processes.

Objective To discuss the effect of royal jelly acid （10-HDA） on the proliferation and migration of the human colon cancer SW620 cells based on the network pharmacology， and to clarify its related molecular mechanism. Methods The active ingredients such as 10-HDA and their corresponding targets were retrieved by using the keyword “royal jelly” from the Traditiomal Chinese Medicine Systems Pharmacology （TMSCP）Database and the Traditiomal Chinese Medicine Integrated Database （TCMID）； the small molecule targets were predicted by the Swiss Target Prediction Database.The GeneCards Database and the Online Mendelian Inheritance in Man（OMIM） Database were used to obtain the targets with the keyword “Colon Cancer”；the protein-protein interaction （PPI） network was constructed by using the String Database and Cytoscape 3.8.0 Software to screen the core targets； the Gene Ontology （GO） function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were analyzed by Metascape Database； the specific ingredient 10-HDA was screened for the in vitro activity experiments. The human colon cancer SW620 cells with good growth status were divided into control group and different doses （1， 5， 10， 15， and 20 mmol·L^－1） of 10-HDA groups. The viabilities of the cells in various groups were detected by MTT method and the survival rates of the cells were calculated. The SW620 cells were divided into control group， low dose （5 mmol·L^－1） of 10-HDA group， middle dose （10 mmol·L^－1） of 10-HDA group， and high dose （15 mmol·L^－1） of 10-HDA group； Hoechst33342 staining method was used to observe the morphology of the cells in various groups； cell scratch test was used to detect the scratch healing rates of the cells in various groups；flow cytometry was used to detect the percentages of the cells at different cell cycles in various groups； biochemical method was used to detect the activities of total antioxidant capacity （T-AOC） and superoxide dismutase （SOD） in the cells in various groups； Western blotting method was used to detect the expression levels of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax），cysteine-containing aspartate proteolytic enzyme-3 （Caspase-3）， cysteine-containing aspartate proteolytic enzyme-9 （Caspase-9）， glycogen synthase kinase 3β （GSK3β）， β-catenin， and cyclin D1 proteins in the cells in various groups. Results Six active ingredients of royal jelly were screened out by the TCMSP Database， and 28 core targets of 10-HDA in the treatment of colon cancer were obtained. The GO function enrichment analysis mainly included the signaling pathways such as cell proliferation and apoptosis. The KEGG signaling pathway enrichment analysis included the cell cycle， prostate cancer， cell senescence， and p53 signaling pathways； the GSK3β/β-catenin signaling pathway was closely related to the cell cycle. Compared with control group， the viabilities of the cells in 5，10，15， and 20 mmol·L^－110-HDA groups were decreased in a dose-dependent manner （P<0.05 or P<0.01）， the numbers of apoptotic cells in different doses of 10-HDA groups were significantly increased， and the scratch healing rates of the cells were significantly decreased （P<0.05 or P<0.01）； the percentages of the cells at S phase in middle and high doses of 10-HDA groups were significantly increased （P<0.05 or P<0.01），the activities of T-AOC and SOD in the cells in different doses of 10-HDA groups were significantly decreased （P<0.05 or P<0.01）. Compared with control group， the expression level of Bcl-2 protein in the cells in low dose of 10-HDA group was significantly decreased （P<0.01）， and the expression level of GSK3β protein was significantly increased （P<0.05）； compared with control group， the expression levels of Bax， Caspase-3， Caspase-9， and GSK3β proteins in the cells in middle and high doses of 10-HDA groups were significantly increased （P<0.05 or P<0.01）， and the expression levels of Bcl-2， β-catenin，and CyclinD1 proteins were significantly decreased （P<0.01）. Conclusion 10-HDA can significantly inhibit the proliferation and migration of the colon cancer cells and promote the apoptosis and oxidation levels of the colon cancer cells，and its mechanism may be related to the activation of the GSK3β / β-catenin signaling pathway.

Objective To confirm the potential etiological factors of congenital aortic stenosis （AS） by genetic analysis on prenatal diagnostic results of the fetus with AS. Methods Amniocentesis for chromosomal G-band karyotyping combinated with single nucleotide polymorphism array （SNP-array） analysis was conducted on the amniotic fluid collected from a 25-week pregnant woman diagnosed as “fetus AS”； chromosome karyotyping was also performed on the peripheral blood of the fetal parents. Results The fetal karyotype analysis showed a chimeric Y-chromosome isobaric double-adherent granules. The SNP-array analysis results revealed a 11.2 Mb duplication in the Yp11.31q11.21 region and a 14.8 Mb deletion in the Yq11.21q11.23 region. Both the parents presented a normal karyotype， suggesting it was a newfound mutation. After extensive genetic counseling， the pregnant woman and her family chose to terminate the pregnancy locally. Conclusion The chromosomal karyotype of the chimeric Y-chromosome isobaric double-adherent granules may be a contributing factor to the AS phenotype in the male fetus. The combined use of chromosomal karyotyping and SNP-array analysis on the amniotic cells is instrumental in the early diagnosis of the disease.

The gastrointestinal motility disorder of the patients with Parkinson’s disease (PD) occurs in the early stages of this disease, even before the onset of motor symptoms. The gastrointestinal motility disorder is one type of gastrointestinal dysfunction, not only affect the absorption of medication, exacerbating the progression of PD, but also severely impact the quality of life of the patients. Therefore, it is essential to find new therapeutic targets to alleviate PD-induced gastrointestinal dysmotility in order to improve the progression of the disease and the quality of life of the patients. The gastrointestinal motility function is highly dependent on the health of the gut and central nervous regulating the gastrointestinal movements. A healthy gut is closely related to the integrity of the intestinal barrier, gut microbiota, neuroinflammation, and the normal function of enteric neurons responsible for the contraction and relaxation of the gastrointestinal tract. The gut function of the PD patients is compromised to some extent. This review summarizes the effects of the enteric nervous system, central nervous system, and gut microbiota on the development of gastrointestinal motility disorder of the PD patients;it also outlines the current therapeutic methods available and their limitations, with the aim of providing the new insights into the treatment of gastrointestinal motility disorder of the PD patients.

Objective To analyze the efficacy of anterior cervical Hybrid surgery and posterior cervical expansive open-door laminoplasty （EODL） in the treatment of multilevel cervical spondylotic myelopathy， and to discuss the selection of surgical methods for the patients with multilevel cervical spondylotic myelopathy. Methods The retrospective analysis was conducted of 70 patients with multilevel cervical spondylotic myelopathy who underwent surgery at Affilated Beijing Traditional Chinese Medicine Hospital of Capital Medical University from July 2017 to July 2020. Based on the different surgical methods， the patients were divided into anterior group （n=35 ）and posterior group（n=35）. The patients in anterior group underwent Hybrid surgery ［anterior cervical discectomy and fusion （ACDF） combined with artificial cervical disc replacement （ACDR）］，and the patients in posterior group underwent EODL. The hospitalization time， operation time， intraoperative blood loss， and postoperative drainage volume of the patients in two groups were recorded； the efficacy was evaluated by Japanese orthopaedic association （JOA） score， JOA improvement rate， neck disability index （NDI）， visual analogue scale （VAS） for pain， and postoperative satisfaction score； the complications of the patients in two groups after surgery were recorded. Results Compared with posterior group， the intraoperative blood loss， postoperative drainage volume， hospitalization time， and operation time of the patients in anterior group were significantly decreased （P<0.01）， and the preoperative score had no significant difference （P>0.05）. At the final follow-up after surgery， compared with posterior group， the JOA score and JOA improvement rate of the patients in anterior group were significantly increased （P<0.01）， and the NDI score and VAS score were significantly decreased （P<0.01）.Compared with before surgery， the JOA scores of the patients in two groups at the final follow-up after surgery were increased （P<0.01）， and the NDI and VAS scores were significant decreased （P<0.01）. The postoperative satisfaction of the patients in two groups was high based on the postoperative satisfaction score.There was no significant difference in the incidence of postoperative complication of the patients between two groups （P>0.05）. Conclusion Both the anterior cervical Hybrid surgery and EODL achieve the satisfactory results in the treatment of multilevel cervical spondylotic myelopathy. Hybrid surgery has the advantages of less bleeding and shorter surgery time， and the most suitable surgical method should be chosen clinically based on the actual situation of the patients.

Objective To discuss the effect of human umbilical cord mesenchymal stem cells culture supernatant （hUCMSCs-Sup） on the proliferation， apoptosis， and endometrium receptivity of the human endometrial stromal cells （hEndoSCs） treated with mifepristone （Ms）， and to clarify the possible mechanism. Methods The hEndoSCs were cultured in vitro and divided into control group and 40， 60， 80， and 100 μmol·L^－1 Ms groups. The survival rates of the cells in various groups were detected by MTT assay. The hEndoSCs were divided into control group， 40 μmol·L^－1 Ms group， and 60 μmol·L^－1 Ms group.The apoptotic rates of the cells in various groups were detected by flow cytometry； the expression levels of apoptosis-related protein B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） proteins in the cells in various groups were detected by Western blotting method， and the ratio of Bcl-2/Bax was calculated. After treated with hUCMSCs-Sup， the hEndoSCs were divided into control group， Ms group， Ms+hUCMSCs-Sup group， and Ms+hUCMSCs-Sup+3-methyladenine （3-MA） group.The survival rates of the cells in various groups were detected by MTT assay；the apoptotic rates of the cells in various groups were detected by flow cytometry；the expression levels of microtubule-associated protein 1 light chain 3B-Ⅱ （LC3B-Ⅱ） and microtubule-associated protein 1 light chain 3B-I （LC3B-Ⅰ） proteins in the cells in various groups were detected by Western blotting method， and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated； the expression levels of endometrium receptivity marker molecules mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. Results Compared with control group， the survival rates of the cells in 40， 60， 80， and 100 μmol·L^－1 Ms groups were significantly decreased （P<0.05）in a time-dependent and dose-dependent manner. Compared with control group， the apoptotic rates of the cells in 40 and 60 μmol·L^－1 Ms groups were significantly increased （P<0.05）， and the ratios of Bcl-2/Bax were significantly decreased （P<0.05）. After treated with hUCMSCs-Sup， compared with control group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms group were significantly decreased （P<0.05）， the apoptotic rate was significantly increased （P<0.05）， and the expression levels of homeobox A10 （HOXA10）， leukemia inhibitory factor （LIF）， and integrin subunit beta 3 （ITGB3） mRNA in the cells were significantly decreased （P<0.05）；compared with Ms group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup group were significantly increased （P<0.05），the apoptotic rate was significantly decreased （P<0.05）， and the expression levels of HOXA10， LIF， and ITGB3 mRNA in the cells were significantly increased （P<0.05）； compared with Ms+hUCMSCs-Sup group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup+3-MA group were significantly decreased （P<0.05）. Conclusion hUCMSCs-Sup can increase the survival rate and decrease the apoptotic rate of the hEndoSCs after treated with Ms，and increase the endometrium receptivity，and its mechanism may be associated with the activation of autophagy of the hEndoSCs by hUCMSCs-Sup.

Objective To discuss the improvement effect of curcumin combined with fecal bacteria transplantation on the mice with dextran sulfate sodium （DSS）-induced ulcerative colitis （UC），and to clarify the related mechanism. Methods Fifty mice were randomly divided into control， model， curcumin， fecal bacteria transplantation， and combination groups. Except for the mice in control group （given distilled water），the mice in the other groups were given distilled water containing 2% DSS to establish the UC models. The mice in curcumin group were gavaged with 0.4 mL of 60 mg·kg^－1 curcumin solution once per day for 10 d； the mice in fecal bacteria transplantation group underwent enema with 0.2 mL of fecal bacteria suspension once per day for 10 d； the mice in combination group received the enema of 0.2 mL fecal bacteria suspension and gavaged with 0.4 mL of 60 mg·kg^－1 curcumin solution. At the end of the experiment， the disease activity index （DAI） and colon macroscopic damage index （CDMI） of the mice in various groups were calculated； the morphology of colon tissue of the mice in various groups was detected by HE staining；the levels of interleukin （IL）-1β， tumor necrosis factor-α（TNF-α）， IL-4， and IL-10 in colon tissue of the mice in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method；the expression levels of occludin and zonula occludens-1 （ZO-1） mRNA and proteins in colon tissue of the mice in various groups were detected by real-time fluorescence quantitative（RT-qPCR） and Western blotting methods. Results The intestinal mucosal epithelial structure of the mice in control group was intact and continuous with regular glandular arrangement and without inflammatory cell infiltration or ulceration； the intestinal mucosal epithelial structure of the mice in model group exhibited loss of colonic mucosal epithelium， disordered glandular arrangement， reduced goblet cells， congestion and edema in mucosal and submucosal layers， and extensive infiltration of inflammatory cells with widespread small ulcers； the intestinal mucosal epithelial structure of the mice in curcumin， fecal bacteria transplantation， and combination groups exhibited relatively intact colonic mucosal epithelial structures with reduced inflammatory cell infiltration and ameliorated mucosal and submucosal congestion and edema. Compared with control group， the DAI and CDMI of the mice in model group were increased （P<0.05）， the levels of IL-1β and TNF-α were increased （P<0.05）， the levels of IL-4 and IL-10 were decreased （P<0.05），and the expression levels of occludin and ZO-1 mRNA and proteins were decreased （P<0.05）；compared with model group， the DAI and CDMI of the mice in curcumin， fecal bacteria transplantation， and combination groups were decreased （P<0.05），the levels of IL-1β and TNF-α were decreased （P<0.05）， the levels of IL-4 and IL-10 were increased （P<0.05）， and the expression levels of occludin and ZO-1 mRNA and proteins were increased （P<0.05）. Compared with curcumin group and fecal bacteria transplantation group，the DAI and CDMI of the mice in combination group were decreased （P<0.05），the levels of IL-1β and TNF-α were decreased （P<0.05）， the levels of IL-4 and IL-10 were increased （P<0.05）， and the expression levels of occludin and ZO-1 mRNA and proteins were increased （P<0.05）. Conclusion Curcumin combined with fecal bacteria transplantation can ameliorate the pathological damage in colonic tissue of the UC mice， inhibit the secretion of inflammatory factors， and promote the repaiment of intestin mucosa.

Objective To discuss the distinctive sonographic feature and the biological behavior of renal angiomyolipoma （RAML）， and to provide the reference for the clinicians to make the accurate diagnosis of RAML. Methods The clinical data of one patient with invasive classical RAML combined with pseudaneurysm formation were collected. The sonographic appearances were analyzed in conjunction with the pathological characteristics to clarify the biological behavior of RAML， and the relevant literatures were reviewed. Results The patient， a 60-year-old female， visited the local hospital due to discomfort in the lumbar area，and received CT examination，and the CT examination results revealed a left renal mass，so the patient came to our hospital. The specialist clinical examinations and laboratory investigations were unremarkable. The ultrasound results indicated an enlarged left kidney with a cystic and solid mass at the upper pole， which featured pseudaneurysm formation （originating from the interlobar arteries）； the enhanced CT image results suggested a high probability of upper pole renal carcinoma combined with aneurysmal formation within the tumor， alongside invasion into the left adrenal gland. The patient underwent laparoscopic radical left nephrectomy， and the postoperative pathology confirmed the diagnosis of invasive classical RAML. Conclusion The classical RAML can exhibit the invasive biological behavior. The pseudaneurysm formation is a special sonographic manifestation of RAML， which can be challenging to differentiate from the other renal tumors.

Objective To discuss the protective effect of gingerone on the hippocampal neuron HT22 cells after oxygen-glucose deprivation/reoxygenation （OGD/R），and to clarify the related mechanism. Methods The HT22 cells were cultured， and the OGD/R cell injury model was established by setting the gradient of OGD/R time. The HT22 cells were divided into control group，OGD/R group， OGD/R+1 μmol·L^－1 gingerone group， OGD/R + 10 μmol·L^－1 gingerone group， OGD/R+100 μmol·L^－1 gingerone group，and OGD/R+0.2% dimethyl sulfoxide（DMSO） group.The viability of the cells in various groups was detected by CCK-8 assay； the survival rates of the cells in various groups were calculated to determine the optimal drug concentration of gingerone. The cells were divided into control， OGD/R group， OGD/R+ gingerone， and OGD/R+gingerone+nuclear factor erythroid-2-related factor 2（Nrf2） inhibitor（ML385） groups.The cells in OGD/R + gingerone group were treated with gingerone for 4 h before OGD treatment for 8 h followed by reoxygenation for 8 h， and the cells in OGD/R+gingerone+ML385 group were treated with 10 μmol·L^－1 ML385 for 6 h before gingerone treatment. The viability of the cells in various groups was detected by CCK-8 assay；the expression levels of Nrf2， heme oxygenase-1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） proteins in the cells in various groups were detected by Western blotting method；the activity of superoxide dismutase （SOD） and the level of malondialdehyde （MDA） in the cell culture supernatant in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method. Results Compared with control group， the survival rate of the HT22 cells was below 50% after treated with OGD for 8 h and reoxygenation for 8 h， so the HT22 cell OGD/R model was established by treated with OGD for 8 h and reoxygenation for 8 h. Compared with OGD/R group，the survival rates of the cells in OGD/R+different doses of gingerone groups were increased to various extents， and the survival rate of the cells in OGD/R+ 100 μmol·L^－1 gingerone group was significantly increased （P<0.01）； so 100 μmol·L^－1 gingerone was used for the subsequent experiment. Compared with control group， the viability of the cells in OGD/R group was significantly decreased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bax proteins in the cells were significantly increased （P<0.01）， while the expression level of Bcl-2 protein in the cells was significantly decreased （P<0.05）， and the SOD activity in the cell culture supernatant was significantly decreased （P<0.01）， and the level of MDA was significantly increased （P<0.01）； compared with OGD/R group， the viability of the cells in OGD/R + gingerone group was significantly increased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bcl-2 proteins in the cells were significantly increased （P<0.05 or P<0.01）， while the expression level of Bax protein in the cells was decreased（P<0.05）， the SOD activity in the cell culture supernatant was significantly increased （P<0.01）， and the level of MDA was significantly decreased （P<0.01）； compared with OGD/R + gingerone group， the viability of the cells in OGD/R + gingerone + ML385 group was significantly decreased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bcl-2 proteins were significantly decreased （P<0.01）， while the expression level of Bax protein in the cells was significantly increased （P<0.01），the SOD activity in the cell culture supernatant was significantly decreased （P<0.01）， and the level of MDA was significantly increased （P<0.05）. Conclusion Gingerone alleviates the oxidative stress damage，and thereby plays an inhibiory effect on the apoptosis of the HT22 neurons by activating the Nrf2/HO-1 signaling pathway after OGD/R.

Objective To discuss the differential effects of apolipoprotein E （APOE） gene polymorphism in the neurotoxicity-reactive astrocytes， and to provide the theoretical basis for the study of the pathogenesis of Alzheimer’s disease （AD）. Methods The primary cortical astrocytes from the APOE-knockout mice （APOE ^-/- ） were isolated and cultured in vitro，and the purity of the cells was identified by immunofluorescence staining. The human APOE3 and APOE4 recombinant over-expression plasmids were constructed and separately transfected into the primary APOE ^-/- astrocytes， and the APOE ^-/- primary cells were regarded as control. Western blotting method was used to detect the expression levels of APOE and glial fibrillary acidic protein （GFAP） proteins in the cells； enzyme-linked immunosorbent assay （ELISA） method was used to detect the APOE level in the cellular culture supernatant. The inflammatory models were prepared with the primary astrocytes transfected with APOE3 and APOE4 and co-stimulated with interleukin-1α（IL-1α）， tumor necrosis factor （TNF）， and complement C1q.The cells were divided into APOE3+PBS group， APOE4+PBS group， APOE3+IL-1α+TNF+C1q group， and APOE4+IL-1α+TNF+C1q group. Cell immunofluorescence staining method was used to observe the morphology of the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of glypican 4 （Gpc4）， glypican 6 （Gpc6）， thrombospondin 1 （Thbs1）， thrombospondin 2 （Thbs2）， SPARC-like protein 1 （Sparcl1） and glial cell line derived neurotrophic factor （GDNF）， C3，and S100 calcium binding protein B （S100B） mRNA in the cells in various groups； microsphere phagocytosis assay was used to detect the phagocytic capacities of the cells in various groups； Western blotting was used to detect the protein expression levels of B-cell lymphoma 2 （Bcl-2），and cysteinyl aspartate specific protease-3 （Caspase-3） proteins in the cells in various groups. Results Compared with APOE ^-/- group， the expression levels of APOE and GFAP proteins in the cells and the APOE level in the cellular culture supernatant in transfected APOE3 and transfected APOE4 groups were increased （P<0.01）. The fluorescence microscope observation results showed that compared with APOE3+PBS and APOE4+PBS groups， the astrocytic processes in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group became shorter and the cell bodies became larger； compared with APOE3+IL-1α+TNF+Cq1 group， the astrocytic processes in APOE4+IL-1α+TNF+Cq1 group were even shorter. Compared with APOE3+PBS and APOE4+PBS groups， the expression levels of Gpc4， Gpc6， Thbs1， Thbs2，and Sparcl1 mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were significantly decreased （P<0.01）； compared with APOE3+IL-1α+TNF+Cq1 group， the expression levels of Gpc4， Gpc6，Thbs1， Thbs2， and Sparcl1 mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group were significantly decreased （P<0.05 or P<0.01）. Compared with APOE3+PBS and APOE4+PBS groups， the expression levels of GDNF mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were decreased（P<0.01），and the expression levels of C3 and S100B mRNA were increased（P<0.01）；compared with APOE3+IL-1α+TNF+Cq1 group， the expression level of GDNF mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased（P<0.05），and the expression levels of C3 and S100B mRNA were increased（P<0.05）. Compared with APOE3+PBS group and APOE4+PBS group，the numbers of hagocytosis of microspheres in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were significantly decreased； compared with APOE3+IL-1α+TNF+Cq1 group，the number of hagocytosis of microspheres in the cells in APOE4+IL-1α+TNF+Cq1 group was significantly decreased. Compared with APOE3+PBS group and APOE4+PBS group，the expression levels of Bcl-2 protein in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were decreased （P<0.05 or P<0.01）and the expression levels of Caspase-3 protein were significantly increased（P<0.01）；compared with APOE3+IL-1α+TNF+Cq1 group， the expression level of Bcl-2 protein in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased（P<0.01），and the expression level of Caspase-3 protein was increased（P<0.05）. Conclusion The APOE4 genotype has a stronger ability to induce the inflammatory factors compared with APOE3；it can lead to a neurotoxicity-reactive astrocyte phenotype，increase the neurotoxicity，affect the astrocyte apoptosis， and aggravate the neuron damage.

Objective To discuss the effect of ligustilide on the cardiac function and angiogenesis in the rats with heart failure，and to clarify its regulatory effect on protein kinase D1 （PKD1）/hypoxia-inducible factor-1α（HIF-1α）/vascular endothelial growth factor （VEGF） pathway. Methods The SD rats were randomly divided into sham operation group， model group， ligustilide group， PKD1/HIF-1α/VEGF signaling pathway inhibitor CID755673 （CID） group， and ligustilide+CID group. The heart failure rat model was established by ligation of the left anterior descending coronary artery. The rats in ligustilide group were injected intravenously with 20 mg·kg^－1 ligustilide， the rats in CID group were injected intraperitoneally with 50 mg·kg^－1 CID， and the rats in ligustilide+CID group were injected intraperitoneally with 50 mg·kg^－1 CID followed by intravenous injection of 20 mg·kg^－1 ligustilide， once per day for 4 consecutive weeks.The cardiac function indexes of the rats in various groups were detected by echocardiography；the percentages of myocardial infarction areas of the rats in various groups were detected by 2，3，5-triphenyltetrazolium chloride （TTC） staining； the pathomorphology of myocardium tissue of the rats in various groups was observed by HE staining； the expression levels of PKD1， HIF-1α， CD31， and VEGF mRNA and proteins in ischemic area of myocardium tissue of the rats in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. Results Compared with sham operation group， the rats in model group and CID group had altered myocardial cell morphology， increased intercellular gaps， disorganized arrangement， visible muscle fiber breaks and inflammatory cell infiltration； the rats in ligustilide group and ligustilide+CID group had relatively orderly myocardial fiber arrangement， fewer myocardial fiber breaks and decreased number of inflammatory cells. Compared with sham operation group， the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） of the rats in model group were decreased （P<0.05）， the left ventricular end-systolic diameter （LVESD） and left ventricular end-diastolic diameter （LVEDD） were increased （P<0.05）， and the expression levels of PKD1， HIF-1α， CD31，and VEGF mRNA and proteins in myocardium tissue were decreased （P<0.05）. Compared with model group， the LVEF and LVFS of the rats in ligustilide group were increased （P<0.05）， the LVESD and LVEDD were decreased （P<0.05），the percentage of myocardium infarction area was decreased （P<0.05）， and the expression levels of PKD1， HIF-1α，CD31， and VEGF mRNA and proteins in myocardium tissue were increased （P<0.05）； compared with model group，the LVEF and LVFS of the rats in CID group were decreased （P<0.05）， the LVESD and LVEDD were increased （P<0.05）， the percentage of myocardium infarction area was increased （P<0.05）， and the expression levels of PKD1， HIF-1α， CD31， and VEGF mRNA and proteins in myocardium tissue were decreased （P<0.05）； compared with ligustilide group， the LVEF and LVFS of the rats in ligustilide+CID group were decreased （P<0.05）， the LVESD and LVEDD were increased （P<0.05）， the percentage of myocardium infarction area was increased （P<0.05）， and the expression levels of PKD1， HIF-1α， CD31， and VEGF mRNA and proteins in myocardium tissue were decreased （P<0.05）；compared with CID group， the LVEF and LVFS of the rats in ligustilide+CID group were increased （P<0.05）， the LVESD and LVEDD were decreased （P<0.05）， the percentage of myocardium infarction area was decreased（P<0.05）， and the expression levels of PKD1， HIF-1α， CD31， and VEGF mRNA and proteins in myocardium tissue were increased （P<0.05）. Conclusion Ligustilide can promote the angiogenesis， reduce the myocardium infarction area， and improve the cardiac function in the rats with heart failure；it works through activation of the PKD1/HIF-1α/VEGF pathway.

Objective To discuss the regulatory effect of berberine （BBR） on fatty acids in the human glioma T98G cells and its effect on the cell proliferation， migration， and invasion， and to clarify its potential mechanism. Methods The T98G cells at logarithmic growth phase were divided into control group and different concentrations （25， 50， and 100 mg·L^－1） of BBR groups. Cell wound healing assay was used to detect the migration rates of the cells in various groups； Transwell chamber assay was used to detect the invasion rates of the cells in various groups.The T98G cells at logarithmic growth phase were divided into control group and 100 mg·L^－1BBR group，and Mass spectrometry was used to detect the fatty acid contents in the cells in two groups. The T98G cells at logarithmic growth phase were divided into control group and different concentrations （50， 100， and 150 mg·L^－1） of BBR groups； Western blotting method was used to detect the expression levels of phosphatidylinositol 3-kinase （PI3K）， phosphorylated PI3K （p-PI3K）， protein kinase B （AKT）， phosphorylated AKT （p-AKT）， sterol regulatory element-binding protein 1 （SREBP-1），and fatty acid synthase （FASN） in the cells in various groups.The expression of FASN was suppressed by gene silencing technology， and the T98G cells at logarithmic growth phase were divided into control group， shFASN1 group， and shFASN2 group. Western blotting method was used to detect the expression levels of FASN protein in the cells in various groups； clone formation assay was used to detect the clone formation of the cells in various groups；cell wound healing assay was used to detect the migration rates of the cells in various groups. Results Compared with control group， the migration rates and invasion rates of the cells in different concentrations of BBR groups were decreased in a concentration-dependent manner （P<0.01）， and the fatty acid content in the cells in 100 mg·L^－1 BBR group was significantly decreased （P<0.01）.Compared with control group，the expression levels of p-PI3K，p-AKT， SREBP-1， and FASN proteins in the cells in 150 mg·L^－1 BBR group were significantly decreased （P<0.05 or P<0.01）， and the expression level of SREBP-1 protein in the cells in 100 and 150 mg·L^－1 BBR groups were significantly decreased （P<0.01）. After suppression of FASN expression， compared with control group， the expression levels of FASN protein in the cells in shFASN1 and shFASN2 groups were significantly decreased （P<0.01）， and the expression level of FASN protein in the cells in shFASN2 group was lower than that in shFASN1 group （P<0.05）； compared with control group， the numbers of clone formation and migration rates of the cells in shFASN1 and shFASN2 groups were significantly decreased （P<0.01）， and the migration rate of the cells in shFASN2 group was significantly lower than that in shFASN1 group （P<0.05）. Conclusion BBR interferes with fatty acid synthesis in the glioma T98G cells by reducing the expression of the PI3K/AKT/SREBP-1/FASN pathway related proteins， and decrease their migration and invasion capabilities.

Objective To discuss the inhibitory effect of chelerythrine （CHE） on the migration， invasion， and epithelial-mesenchymal transition （EMT） of the human ovarian cancer SKOV3 cells，and to clarify the associated mechanism. Methods The SKOV3 cells were cultured in vitro and divided into control group and 2.5， 5.0， 10.0， 20.0， and 40.0 μmol·L^－1 CHE groups.Methylthiazolydiphenyl-tetrazolium（MTT） assay was used to detect the inhibitory rates of proliferation of the cells in various groups. The SKOV3 cells were cultured in vitro and divided into control group， transforming growth factor-β1 （TGF-β1） group， TGF-β1+5 μmol·L^－1 CHE group， and TGF-β1+10 μmol·L^－1 CHE group.Cell scratch assay was used to detect the migration rates of the cells in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups； Western blotting method was used to detect the expression levels of E-cadherin， N-cadherin， and Vimentin proteins in the cells in various groups； immunofluorescence staining method was used to detect the fluorescence intensities of E-cadherin and N-cadherin in the cells in various groups. Results The MTT assay results showed that compared with control group， the inhibitory rates of proliferation of the cells in 5.0， 10.0， 20.0， and 40.0 μmol·L^－1 CHE groups were significantly increased （P<0.05 or P<0.01）. The cell scratch assay results showed that compared with control group， the migration rate of the cells in TGF-β1 group was increased （P<0.01）； compared with TGF-β1 group， the migration rates of the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with control group， the numbers of migration and invasion cells in TGF-β1 group were significantly increased （P<0.05）； compared with TGF-β1 group， the numbers of migration and invasion cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of E-cadherin protein in the cells in TGF-β1 group was significantly decreased （P<0.01）， while the expression levels of N-cadherin and Vimentin proteins were increased （P<0.05 or P<0.01）； compared with TGF-β1 group， the expression levels of E-cadherin protein in the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly increased （P<0.01）， and the expression levels of N-cadherin and Vimentin proteins were significantly decreased （P<0.01）. The immunofluorescence staining results showed that compared with control group， the fluorescence intensity of E-cadherin in the cells in TGF-β1 group was decreased， and the fluorescence intensity of N-cadherin was increased； compared with TGF-β1 group， the fluorescence intensities of E-cadherin in the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly increased， and the fluorescence intensities of N-cadherin were decreased. Conclusion CHE can inhibit the proliferation， migration， invasion， and EMT of the human ovarian cancer SKOV3 cells.

Objective To analyze the clinical presentations and diagnostic and treatment process of one patient with autoimmune encephalitis（AE） with double positive anti-N-methyl-D-aspartate receptor （NMDAR） and anti-γ-aminobutyric acid B receptor （GABA_BR） secondary to herpes simplex virus encephalitis（HSVE），and to improve the clinicians’ awareness of this disease. Methods The clinical data of one AE patient with double positive anti-NMDAR and anti-GABA_BR secondary to HSVE were collected， the diagnostic and therapeutic processes were summarized， and the relevant literatures were reviewed. Results The patient， a 36-year-old male， developed a headache followed by limb convulsions， and progressed to disturbed consciousness. After admission， the routine biochemistry of the cerebrospinal fluid （CSF） was abnormal， and the herpes simplex virus-1 （HSV-1） IgG antibody showed positive in the CSF； both CSF and serum tests for NMDAR antibodies were positive； the head magnetic resonance imaging （MRI） results showed abnormal signals in the right occipital white matter， leading to the diagnosis of HSVE secondary to anti-NMDAR encephalitis. Several months later， the patient experienced psychiatric behavior abnormalities， cognitive dysfunction， and sleep disorders， and both the serum NMDAR and GABA_BR antibodies showed positive results， prompting the diagnosis of HSVE secondary anti-NMDAR encephalitis and anti-GABA_BR encephalitis. After treatment with steroid pulse therapy and intravenous immunoglobulin （IVIG）， the patient’s condition was improved and the patient was discharged. At one-year follow-up， the patient’s psychiatric symptoms had completely resolved， leaving mild cognitive impairment. Conclusion If the clinical symptoms of the patients recovering from antiviral treatment for HSVE is worsened， secondary AE should be highly suspected；it is important to complete autoimmunity antibody testing as soon as possible for the early diagnosis and treatment to improve the prognosis of the patient.

Objective To discuss the clinical characteristics， diagnosis processes， and treatment methods of one patient with congenital intrabdominal hernia，and to summarize the potential misconceptions during the diagnostic and treatment processes， and to improve the clinicians’ awareness of this disease. Methods The clinical data and auxiliary examination results of one patient with congenital intrabdominal hernia were collected and analyzed， and the related literatures were reviewed. Results The patient， a 65-year-old male， sought care at the local hospital due to upper abdominal pain before 2 d；there were no significant abnormalities in the examination results at the cocal hospital；blood glucose>25 mmol·L^－1.After receiving hypoglycemic， rehydration， and blood purification treatment， the condition of the patient was worsened， presenting with confusion， hypotension， and respiratory distress；the patient admitted in our hospital for further diagnosis and treatment.After admission，the patient was given despite fluid resuscitation， mechanical ventilation， and supportive treatment， but there was no improvement in the symptoms；interventional radiology was performed angiography of the abdominal artery and right femoral vein， which showed no significant vascular abnormalities in the abdomen. An abdominal paracentesis yielded a mixed bloody fluid， suggesting the concealed intraperitoneal disease； exploratory laparotomy was performed. During operation， the intrabdominal hernia with small intestine necrosis and septic shock were diagnosed， and partial small intestine resection， anastomosis， adhesiolysis， and abdominal irrigation and drainage were carried out. The patient had a good recovery and was discharged on the 14th day after operation. Conclusion Congenital intrabdominal hernia is a very rare cause of intestinal obstruction in the adults，and high suspicion for intrabdominal hernia is one of the differential diagnosis for atypical acute abdomen；early multidisciplinary intervention can be lifesaving for the patients.

Objective To screen the aging genes closely associated with pelvic organ prolapse （POP） by bioinformatics techniques， and to clarify the potential clinical significance and value of key genes. Methods Gene Expression Omnibus （GEO） Database was used to download the datasets GSE53868 and GSE151188 for POP-related genes with the keyword “pelvic organ prolapse”. The aging-related genes were obtained from Aging Atlas， CellAge， and the Human Ageing Genomic Resources （HAGR） Databases；the intersection of genes related with POP in two groups provided a list of differentially expressed genes （DEGs） associated with aging in POP； gene Set Enrichment Analysis （GSEA） was conducted with R software version 4.2.1； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis of DEGs were conducted by the Database for Annotation， Visualization and Integrated Discovery （DAVID）； the protein-protein interaction （PPI） network was constructed with Cytoscape 3.9.1 software；the top 10 Hub genes were selected by cytoHubba plugin； the infiltration of 22 types of immune cells in the patients in POP group and control group was analyzed by CIBERSORT deconvolution method using R software；the key genes were further screened by LASSO regression algorithm； the correlation and diagnostic efficacy between key genes and immune cell infiltration were analyzed. Results From the Aging Atlas， CellAge， and HAGR Databases， 724 aging-related genes were identified. Intersection with the POP expression profile yielded an aging gene expression matrix related to POP containing 624 genes， and 29 POP-related DEGs were identified after differential analysis， including 2 upregulated genes and 27 downregulated genes. The GSEA results showed that the upregulated pathways were mainly related to diabetes and cellular senescence， whereas the downregulated pathways included Alzheimer’s disease and hypoxia-inducible factor-1 （HIF-1） signaling pathways.The GO functional enrichment analysis mainly enriched in the biological processes such as the response of the cells to lipopolysaccharide， inflammatory response， and negative regulation of cell proliferation. The KEGG signaling pathway enrichment analysis mainly enriched in interleukin-17 （IL-17）， tumor necrosis factor （TNF）， and nuclear factor-kappa B （NF-κB） signaling pathways. The PPI network analysis got 10 Hub genes including interleukin-6 （IL-6）， interleukin-1B （IL-1B）， prostaglandin-endoperoxide synthase 2 （PTGS2）， and NF-kappa-B inhibitor alpha （NFKBIA）. The CIBERSORT deconvolution method results showed a relatively higher infiltration proportion of neutrophils and activated mast cells in the patients in POP group， the activated mast cells had a positive correlation with most of the DEGs （r>0.5） and the macrophages had a significant positive correlation with IL-1B （r>0.6）. The key genes Jun D proto-oncogene （JUND）， Snail homolog 1 （SNAI1）， amphiregulin （AREG）， Lamin A/C （LMNA）， and superoxide dismutase 2 （SOD2） selected by LASSO regression analysis had high diagnostic efficacies， and the area under receiver operating characteristic curve （ROC） （AUC） were all greater than 0.75. Conclusion During the aging process，the genes such as JUND，SNAI1，AREG，LMNA，and SOD2 may participate in the pathophysiology of POP through various pathways，including inflammation-related pathways，transcription regulation，and affecting collagen secretion and metabolism，thereby influence the connective tissue support function and promote the occurrence and development of POP.

Objective To investigate the efficacy of Balanophora involucrata Hook.f. in treatment of hyperuricemia （HUA） based on network pharmacology， molecular docking， and hyperuricemia models in vivo and in vitro，and to clarify the main targets of its active components and related signaling pathway mechanism. Methods The potential targets of Balanophora involucrata Hook.f. in treatment of HUA were identified by Databases such as the Traditional Chinese Medicine Database in Taiwan， the Chinese Herbal Medicine Identification Database，Professional Chemical Database， TargetNet Database， SwissTargetPrediction Database， GeneCards， Therapeutic Target Database （TTD），DrugBank Database， DisGeNET Database， Online Mendelian Inheritance in Man （OMIM） Database， and Venny Database. STRING Database and Cytoscape software were used to construct the active component-predictive target network and protein-protein interaction （PPI） network for Balanophora involucrata Hook.f.；topological analysis was used to select the main active components and core targets；Gene Ontology （GO） functional and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed by R software； AutoDock Vina software was used for molecular docking validation. The NRK-52E cells were divided into blank control group， blank administration group， model group， and different concentrations （2.0， 10.0， and 50.0 μmol·L^-1） of erythrodiol （EDT） groups. High-performance liquid chromatography culture （HPLC） was used to detect the uric acid （UA） levels in the cell culture supernatants in various groups. The male ICR mice were divided into blank control group， blank administration group， model group， and EDT group； the mice in the last two groups were used to prepare the HUA models； kits were used to detect the levels of UA， creatinine （Cr）， and blood urea nitrogen （BUN） in serum of the mice in various groups；the bilateral kidney tissue of the mice was harvested and weighed； the kidney indexes of the mice in various groups were calculated；TUNEL staining was used to observe the apoptosis in kidney tissue of the mice in various groups；Western blotting method was used to detect the expression levels of protein kinase B （AKT），phosphorylated AKT （p-AKT），phosphoinositide 3-kinase （PI3K），phosphorylated PI3K（p-PI3K）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and matrix metalloproteinase-9 （MMP-9） proteins in kidney tissue of the mice in various groups. Results Six active components of Balanophora involucrata Hook.f.were identified， involving 116 intersecting targets and 14 core targets.The enrichment analysis yielded 1 828 GO terms and 145 signaling pathways. The molecular docking results showed that EDT had good binding activity with MMP-9. The high uric acid cell experiment results showed that compared with blank control group， the UA level in the cells in model group was significantly increased （P<0.01）； compared with model group， the UA levels in the cells in 2.0， 10.0， and 50.0 μmol·L^-1 EDT groups were significantly decreased （P<0.01）. Compared with blank control group， the levels of UA， Cr， and BUN in serum of the mice in model group were increased（P<0.01）， and the kidney indexes were significantly increased （P<0.01）； compared with model group， the levels of UA， Cr， and BUN in serum of the mice in EDT group were decreased （P<0.05 or P<0.01），and the kidney index was significantly decreased （P<0.05 or P<0.01）. Compared with blank control group， the number of apoptotic cells in kidney tissue of the mice in model group was increased； compared with model group， the number of the apoptotic cells in kidney tissue of the mice in EDT group was significantly decreased. Compared with blank control group， the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in model group were significantly decreased （P<0.05 or P<0.01）， while the expression levels of Bax and MMP-9 proteins were significantly increased （P<0.01）； compared with model group， the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in EDT group were significantly increased （P<0.05 or P<0.01）， and the expression levels of Bax and MMP-9 proteins were significantly decreased （P<0.01）. Conclusion The active component of Balanophora involucrata Hook.f.，EDT，has a UA-decreasing effect and may inhibit the apoptosis and alleviate the kidney injury by activating the PI3K/AKT signaling pathway.

Objective To screen out the key genes affecting lung adenocarcinoma （LUAD） through bioinformatics methods，and to analyze their biological functions and the influences on the LUAD prognosis. Methods The GSE118370 and GSE136043 chip data were downloaded from the Gene Expression Omnibus（GEO） Database. The LUAD-related data were selected from the The Cancer Genome Atlas（TCGA） Database. R software was used to analyze the co-expressed differentially expressed genes （DEGs）； clusterProfile R package was utilized for Gene Ontology （GO） functional enrichment analysis； DAVID Database was used for the Kyoto Gene and Genome Encyclopedia （KEGG） signaling pathway enrichment analysis； STRING Database was used to construct the protein-protein interaction （PPI） network； Cytoscape was used to screen out the top 10 key genes； GEPIA Database and Human Protein Atlas（HPA） Database were used to analyze the expressions of key genes mRNA and protein in normal lung tissue and LUAD tissue， and their expressions in LUAD tissues with different stages；immune infiltration analysis and survival analysis were used to analyze the correlation between the expressions of key genes and the survival time of the patients. Results In total， 428 DEGs were screened out. The GO functional analysis results showed that the DEGs of LUAD were mainly enriched in biological process（BP） such as epithelial-mesenchymal transition （EMT）， cellular component（CC） such as the cell base， and molecular function（MF） such as extracellular matrix （ECM） structure formation.The KEGG signaling pathway analysis results showed that the DEGs of LUAD were mainly enriched in the pathways like cytokine receptor interactions.topoisomerase Ⅱ alpha（TOP2A），abnormal spindle microtubule assembly（ASPM），cyclin B1，（CCNB1），cell division cycle associated 8（CDCA8），baculoviral IAP repeat containing 5（BIRC5），aurora A（AURKA），kinesin family member 20A（KIF20A），centrosomal protein 55（CEP55），centromere protein F（CENPF），and targeting protein for xklp2（TPX2） were selected as the key genes. Compared with normal lung tissue， the expression levels of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 mRNA in lung tissue of the LUAD patients were increased （P<0.01）， and the expression levels of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 proteins were increased. There were significant differences in the expression levels of CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， and TPX2 mRNA in the LUAD tissue with different stages （P<0.01）. Compared with the LUAD patients with stage Ⅰ，stage Ⅱ，and stage Ⅲ，the expression levels of CCNB1， CDCA8， AURKA， KIF20A， CEP55， and TPX2 mRNA in lung tissue of the LUAD patients with stage Ⅳ were increased（P<0.01）； compared with the LUAD patients with stage Ⅰ，stage Ⅱ， and stage Ⅳ，the expression level of BIRC5 mRNA in lung tissue of the LUAD patients with stage Ⅲ was increased （P<0.01）. The expressions of 10 key genes were negatively correlated with the B-lymphocyte infiltration （－0.253≤r≤－0.014， P<0.01）； the expressions of TOP2A， ASPM， CDCA8， BIRC5， CEP55， CENPF， and TPX2 were positively correlated with the neutrophil infiltration （0.049≤r≤0.165，P<0.01）； the expressions of CCNB1 and AURKA were negatively correlated with the CD4 T lymphocyte， macrophage， and dendritic cell infiltration （－0.210≤r≤－0.100，P<0.01）. The high expression of CDCA8 increased the risk of LUAD deterioration （P<0.01）， and the high expressions of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 increased the death risk of the patients（P<0.01）. Conclusion TOP2A， ASPM， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 are the key genes involved in the development and progression of LUAD. They may promote the development of LUAD by accelerating the EMT process， and their high expressions suggest the poor prognosis and elevate the death risk of the LUAD patients.

Objective To construct the sex-determined region Y-box 17 （SOX17） over-expression lentiviral vector， and to construct the cell line stably over-expressing SOX17 by using the PC12 cells infected with SOX17 over-expression lentivirus. Methods The over-expression sequence of SOX17 was designed and synthesized based on the National Center for Biotechnology Information （NCBI） Database， and was connected to the GV492 lentiviral vector after being doubly digested with BamHⅠ and AgeⅠ enzymes to construct the GV492-SOX17 over-expression recombinant plasmid. The agarose gel electrophoresis was used to identify the PCR products， and the positive bacteria carrying the GV492-SOX17 over-expression recombinant plasmid were selected， cloned and sequenced. The GV492 empty plasmid and GV492-SOX17 over-expression recombinant plasmid were transfected into the human embryonic kidney HEK 293T cells， respectively. After transfected for 48 h， the GV492 control lentivirus and GV492-SOX17 over-expression lentivirus were packaged and the viral titer was detected. The PC12 cells were divided into blank group， GV492 control group， and GV492-SOX17 group. The cells in blank group was not treated， and the cells in GV492 control and GV492-SOX17 groups were infected with the corresponding lentivirus （multiplicity of infection = 100）， and the PC12 cells successfully infected with lentivirus were selected with 10 mg·L^－1 puromycin. The growth state and green fluorescence expression of the PC12 cells were observed under fluorescence microscope；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of SOX17 mRNA in the PC12 cells in various groups；Western blotting method was used to detect the expression level of SOX17 protein in the PC12 cells in various groups. Results The gene fragment length of GV492-SOX17 over-expression recombinant plasmid was about 744 bp， and the sequence of GV492-SOX17 over-expression recombinant plasmid gene was identical to the designed and synthesized SOX17 over-expression sequence. The titers of GV492 control lentivirus and GV492-SOX17 over-expression lentivirus were both 2.5×108 TU·mL^－1. The growth state of the PC12 cells in various groups was good and the green fluorescence was expressed. The RT-qPCR results showed that the expression level of SOX17 mRNA in the cells in GV492-SOX17 group was significantly higher than those in blank group and GV492 control group（P<0.01）. The Western blotting results showed that there was a specific band appeared at a relative molecular mass of 44 000， suggesting the SOX17 protein was successfully expressed in the PC12 cells； compared with blank group and GV492 control group， the expression level of SOX17 protein in the cells in GV492-SOX17 group was increased （P<0.05）. Conclusion The GV492-SOX17 lentiviral expression vector is successfully constructed and the PC12 cell line stably over-expressing GV492-SOX17 is established.

Objective： To analyze the clinical performance and diagnosis process of the autoimmune encephalitis （AE） patients with positive double antibodies in the cerebrospinal fluid（CSF）， and to provide the references for the diagnosis and treatment of such patients. Methods The clinical manifestations， magnetic resonance imaging （MRI） of the head， electroencephalogram （EEG）， CSF characteristics， and prognosis of one patient with AE positive for anti-glutamic acid decarboxylase （GAD） 65 and anti-γ-aminobutyric acid B receptor （GABA_BR） double antibodies in the CSF were retrospectively analyzed，and the literatures were reviewed. Results The patient was a 47-year-old male with subacute onset and progressively aggravated symptoms， mainly presenting with headaches and episodic convulsions， and blurred consciousness.The MRI results of the head suggested that the lesions were located on the right side of the cerebral falx in the frontal and parieto-occipital lobes；the CSF was positive for the anti-GAD65 and anti-GABA_BR double antibodies， and the EEG results showed the abnormal spike and slow waves， so the patient was diagnosed as AE. After anti-inflammatory and other symptomatic treatments， the patient was gradually improved and was discharged；the patient was given continuous oral corticosteroid treatment， but after 5 months， the patient was relapsed with acute onset， presenting with convulsion accompanied by drooling from the corner of the mouth. The head MRI results showed there was an abnormal high signal in the right temporal lobe. After corticosteroid treatment， the patient was improved. Conclusion The AE patients with positive double antibodies in CSF are more likely to relapse. Steroid anti-inflammatory treatment is effective. When intracranial lesions are located in the frontal and parieto-occipital lobes， it should be considered the possibility of symptoms such as convulsions. It is necessary to complete the EEG and the other inspections as soon as possible.

Objective To discuss the protective effect of fructus mume total flavonoids （FMF） on the injury of the SH-SY5Y cells induced by 1-methyl-4-phenylpyridinium（MPP⁺） through regulating microRNA （miR）-145-3p expression，and to clarify the mechanism. Methods The SH-SY5Y cell model of Parkinson’s disease （PD） was established by MPP⁺ induction. The optimal intervention concentration and action time of MPP⁺ and FMF were screened out by CCK-8 assay. The cells were divided into control group （untreated）， model group （treated with 500 μmol·L^－1 MPP⁺ for 24 h）， MPP⁺+mimics group （transfected with miR-145-3p mimics）， MPP⁺+NC group （transfected with miR-145-3p NC）， MPP⁺+FMF group treated with 0.25 g·L^－1 FMF for 24 h）， MPP⁺+mimics+FMF group （transfected with miR-145-3p mimics and treated with 0.25 g·L^－1 FMF for 24 h），and MPP⁺+NC+FMF group （transfected with miR-145-3p NC，and （treated with 0.25 g·L^－1 FMF for 24 h）. CCK-8 assay was used to detect the proliferation abilities of the cells in various groups； Annexin Ⅴ-FITC/PI staining was used to the detect the apoptotic rates of the cells in various groups； transmission electron microscope was used to observe the ultrastructure morphology of mitochondrial autophagy in the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-145-3p in the cells in various groups； Western blotting method was used to detect the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in various groups. Results After treated with different concentrations of MPP⁺， the proliferation ability of the cells was significantly decreased（P<0.01）， and the PD cell model was successfully constructed. The optimal concentrations and action time of MPP⁺ and FMF intervention were 500 μmol·L^－1， 24 h， and 0.25 g·L^－1，24 h， respectively.The CCK-8 assay results showed that the proliferation ability of the cells in model group was significantly lower than that in control group （P<0.01）； the proliferation abilities of the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than that in model group（P<0.01）； the proliferation ability of the cells in MPP⁺+mimics+FMF group was significantly higher than that in MPP⁺+FMF group （P<0.01）. The Annexin V-FITC/PI staining results showed that the apoptotic rate of the cells in model group was significantly higher than that in control group （P<0.01）； the apoptotic rates of the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly lower than that in model group （P<0.01）； the apoptotic rate of the cells in MPP⁺+mimics+FMF group was significantly lower than that in MPP⁺+FMF group （P<0.01）. The transmission electron microscope results showed that the cells in control group showed the uniform and intact mitochondrial morphology， the cells in model group showed the enlarged and irregular mitochondria with vacuolar changes，and the cells in MPP⁺+mimics group and MPP⁺+FMF group showed the improved mitochondrial structure， compared with model group， the autophagosome numbers of the cells in MPP⁺+mimics group and MPP⁺+FMF group were increased； the cells in MPP⁺+mimics+FMF group showed more intact mitochondrial structure， compared with MPP⁺+FMF group， the autophagosome numbers in the cells was increased. The RT-qPCR results showed that the expression level of miR-145-3p in the cells in model group was significantly lower than that in control group （P<0.01）； the expression levels of miR-145-3p in the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than that in model group （P<0.01）； the expression level of miR-145-3p in the cells in MPP⁺+mimics+FMF group was significantly higher than that in MPP⁺+FMF group （P<0.01）.The Western blotting results showed that the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/LC3-Ⅰ in the cells in model group were lower than those in control group （P<0.05）； the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than those in model group （P<0.01）； the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in MPP⁺+mimics+FMF group were significantly higher than those in MPP⁺+FMF group （P<0.01）. Conclusion FMF may promote the autophagy， alleviate the mitochondrial dysfunction， and attenuate the injury of the SH-SY5Y cells induced by MPP⁺， and its mechamism is related to upregulating the miR-145-3p expression.

Objective To discuss the effectiveness of machine learning method combined with magnetic resonance diffusion weighted imaging （DWI） for recognition of the acute ischemic stroke （AIS） patient with onset time within 4.5 h，and to provide the reference for assisted assessment of onset time of AIS patients. Methods A total of 227 AIS patients with complete DWI imaging data were divided into onset time≤ 4.5 h group （n=70） and onset time >4.5 h group （n=157） based on their time from onset to DWI examination. The patients were randomly divided into training set （n=158） and test set （n=69） at a ratio of 7∶3. The regions of interest （ROI） were designated on the DWI images by ITK-SNAP annotation software， and the image features were extracted from the ROI images by the Pyradiomics package.The redundant features with high consistency were removed after evaluating the correlation of each feature by Spearman correlation test. Least absolute shrinkage and selection operator （LASSO） regression model with 10-fold cross-validation was used to recognize the image features of the AIS patients with onset time within 4.5 h. Seven machine learning algorithms， including Support Vector Machines （SVM）， K-Nearest Neighbors （KNN）， Decision Tree， Extra Trees， Random Forest， XGBoost， and LightGBM， were used to train the models. The performance of the models was evaluated by receiver operating characteristic curve （ROC） and the area under the curve （AUC）. Results A total of 107 image features， including 18 first-order features， 14 shape features， and 75 texture features， were extracted from the ROI images， among which 22 features were finally selected for recognition of the AIS patients with onset time within 4.5 h， including 4 first-order features， 6 shape features， and 12 texture features. The XGBoost model yielded the best results in recognizing the AIS patients with onset time within 4.5 h， and the AUC was 0.817， and the accuracy， sensitivity， and specificity were 0.739， 0.733， and 0.814，respectively. Conclusion The XGBoost model based on DWI imaging can effectively recognize the AIS patients with onset time within 4.5 h， which has certain clinical significance in assisting the assessment of onset time of the AIS patients.

Objective To discuss the effect of usnic acid on the proliferation， apoptosis， migration， and invasion of the hypertrophic scar fibroblasts （HSFBs）， and to clarify its regulatory effect on the c-Jun N-terminal kinase （JNK）/mitogen-activated protein kinase （MAPK） signaling pathway. Methods The HSFBs at logarithmic growth phase were divided into control group， 10 μmol·L^－1 usnic acid group， 20 μmol·L^－1 usnic acid group， and 50 μmol·L^－1 usnic acid group. Except for control group， the cells in the other groups were treated with the corresponding concentrations of usnic acid. The survival rates of the cells in various groups were detected by methylthiazolyidiphenyl-tetrazolium bromide（MTT） method； the apoptotic rates of the cells in various groups were detected by flow cytometry；the invasion and migration abilities of the cells in various groups were detected by Transwell chamber experiment；the expression levels of phosphorylated JNK （p-JNK）， JNK， B-cell lymphoma-2（Bcl-2）， and Bcl-2-associated X protein （Bax） proteins in the cells in various groups were detected by Western blotting method. Results The MTT and flow cytometry results showed that compared with control group， the survival rates of the cells in 10， 20， and 50 μmol·L^－1 usnic acid groups were decreased （P<0.05）， and the apoptotic rates were increased （P<0.05）. Compared with 50 μmol·L^－1 usnic acid group， the survival rates of the cells in 10 and 20 μmol·L^－1 usnic acid groups were decreased （P<0.05）， and the apoptotic rates were increased （P<0.05）.The Transwell chamber experiment results showed that compared with control group，the numbers of invasion and migration cells in 10，20 and 50 μmol·L^－1 usnic acid groups were decreased（P<0.05）； compared with 10 μmol·L^－1 usnic acid group，the numbers of invasion and migration cells in 20 and 50 μmol·L^－1 usnic acid groups were decreased（P<0.05）；compared with 20 μmol·L^－1 usnic acid group，the numbers of invasion and migration cells in 50 μmol·L^－1 usnic acid groups were decreased（P<0.05）.The Western blotting results showed that compared with control group，the expression levels of Bcl-2 protein in the cells in 10， 20， and 50 μmol·L^－1 usnic acid groups were decreased（P<0.05）， and the expression levels of Bax and p-JNK proteins were increased（P<0.05）；compared with 10 μmol·L^－1 usnic acid group，the expression levels of Bcl-2 protein in the cells in 20 and 50 μmol·L^－1 usnic acid groups were decreased， and the expression levels of Bax and p-JNK proteins in the cells were increased（P<0.05）； compared with 20 μmol·L^－1 usnic acid group，the expression level of Bcl-2 protein in the cells in 50 μmol·L^－1 usnic acid group was decreased（P<0.05）， and the expression levels of Bax and p-JNK proteins were increased（P<0.05）. Conclusion Usnic acid can inhibit the proliferation， invasion， and migration of the HSFBs， induce the apoptosis of the HSFBs， and activate the JNK/MAPK signaling pathway.

Objective To discuss the effect of polysaccharides from porphyra on the inflammatory factors secreted by macrophage RAW264.7 cells in the mice induced by lipopolysaccharide （LPS），and to clarify their immunomodulatory effect. Methods The RAW264.7 cells at logarithmic growth phase were divided into control group， LPS group， and LPS+polysaccharide from porphyra group. The viabilities of the cells in various groups were detected by CCK-8 method；the levels of interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α）， monocyte chemotactic protein-1 （MCP-1）， macrophage inflammatory protein　（MIP）-1α，and MIP-1β in the cell culture supernatants in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method. Results Compared with control group， the survival rate of the cells and the levels of IL-6， TNF-α， MCP-1， MIP-1α， and MIP-1β in the culture supernatant in LPS group and LPS+polysaccharide from porphyra group were significantly increased （P<0.01）. Compared with LPS group， the survival rate of the cells and the levels of IL-6， TNF-α， MCP-1， MIP-1α， and MIP-1β in the culture supernatant in LPS+polysaccharide from porphyra group were significantly decreased （P<0.01）. Conclusion polysaccharides from porphyra can inhibit the impact of the mouse macrophages on the secretion of inflammatory factors IL-6，TNF-α，MCP-1，MIP-1α，and MIP-1β，and have the potential immunomodulatory effect.

Objective To discuss the effect of ligustrazine on growth of the glioma stem cells（GSCs）subcutaneous xenografts in the nude mice， transforming growth factor-β （TGF-β） signaling pathway and epithelial-mesenchymal transition（EMT），and to provide the reference for selecting drugs in the clinical therapy of glioma. Methods The human brain glioma U87 cells were collected and the GSCs were cultured， separated， enriched， and identified. Forty female BALB/c nude mice were used to establish the GSCs subcutaneous xenograft models， which was randomly divided into model group and low， medium， and high doses of ligustrazine groups， and there were 10 mice in each group. The mice in low， medium， and high doses of ligustrazine groups were intraperitoneally injected with ligustrazine once every 3 d， and the dosages were 1.5， 3.0， and 6.0 mg·kg^－1， respectively， while the mice in model group were only given equal volume of saline，and this process continued for 15 d. The transplanted tumor volumes and tumor weights were detected， and the inhibitory rates of tumor weights of the mice was caculated. The tumor growth curve was drawn.HE staining was used to observe the pathomorphology of tumor tissue of the mice；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of TGF-β1 and transforming growth factor β receptor Ⅰ（TβRⅠ） mRNA in tumor tissue of the mice in various groups； Western blotting method was used to detect the expression levels of TGF-β1， TβRⅠ，small mother against decapentaplegic homolog 2/3 （Smad2/3）， phosphorylated Smad2/3 （p-Smad2/3），E-cadherin， TWIST family bHLH transcription factor 1 （TWIST1），Vimentin， and Snail proteins in tumor tissue of the mice in various groups. Results The GSCs were successfully enriched and separated from the U87 cells， and the immunofluorescence staining resutls showed the positive expression of stem cell marker CD133. Compared with model group， the transplanted tumor volumes of the nude mice in low， medium， and high doses of ligustrazine groups were significantly decreased （P<0.05）， in a dose-dependent manner； the tumor weights were also significantly decreased （P<0.05） in a dose-dependent manner as well. The inhibitory rates of tumor weights in low， medium， and high doses of ligustrazine groups were 12.72%， 39.90%， and 55.36%， respectively.The HE staining results showed that the cells in tumor tissue of the mice in model group were gradually increased and exhibited good morphology and tight connection， the growth density was high， and the atypical nuclei was obviously； the tumor tissue structures of the mice in low， medium， and high doses of ligustrazine groups were disordered， the cell density was gradually decreased， the nuclear condensation and karyorrhexis were increased，the necrotic zones were gradually enlarged， and these improvements became more obvious as the increasing of the dose of ligustrazine. The RT-qPCR results showed that compared with model group， the expression levels of TGF-β1 and TβRⅠ mRNA in tumor tissue of the mice in low， medium， and high doses of ligustrazine groups were decreased （P<0.05） in a dose-dependent manner.The Western blotting results showed that compared with model group， the expression levels of E-cadherin protein in tumor tissue of the mice in low， medium， and high doses of ligustrazine groups were increased （P<0.05），and the expression levels of TWIST1，Vimentin，Snail，TGF-β1，TβRⅠ，and p-Smad2/3 proteins were decreased（P<0.05） in a dose-dependent manner. Conclusion Ligustrazine can inhibit the growth of GSCs subcutaneous xenografts in the nude mice， and can interfere the TGF-β1/Smad2/3 signaling activation and EMT induction.

Objective To discuss the change of reticulocyte hemoglobin （RET-He） level of the anemic patients with different degrees and different types of anemia， and to analyze its auxiliary diagnostic value in microcytic hypochromic anemia. Methods The clinical data of 369 anemic patients and 40 healthy physical examination individuals were collected. The patients were divided into mild anemia group （125 cases）， moderate anemia group （163 cases）， and severe anemia group （81 cases） based on the hemoglobin （Hb） level，and 40 healthy physical examination individuals were regarded as control group. The patients were also divided into macrocytic anemia group （91 cases）， normocytic anemia group （108 cases）， simple microcytic anemia group （23 cases）， and microcytic hypochromic anemia group （147 cases） based on the morphological types of anemia. The red blood cell count， Hb level， blood cell specific volume （HCT）， mean red blood cell volume （MCV）， mean red blood cell Hb level （MCH），mean red blood cell Hb concentration （MCHC）， and RET-He level of the subjects in various groups were detected；the receiver operating characteristic （ROC） curve was drawn to obtain the diagnostic cutoff value for the disease； the area under curve （AUC）， sensitivity， and specificity were calculated. Results Compared with control group， the RET-He levels of the patients in moderate and severe anemia groups were significantly decreased （P<0.01）； compared with mild anemia group， the RET-He levels of the patients in moderate and severe anemia groups were significantly decreased（P<0.01）. The RET-He levels of the patients in macrocytic anemia， normocytic anemia， simple microcytic anemia， and microcytic hypochromic anemia groups showed a decreasing trend （P<0.01）； compared with control group， the RET-He levels of the patients in simple microcytic anemia and microcytic hypochromic anemia groups were significantly decreased （P<0.01）； compared with mild macrocytic anemia group， the RET-He levels of the patients in mild normocytic anemia， mild simple microcytic anemia， and mild microcytic hypochromic anemia groups were significantly decreased （P<0.01）. Compared with mild normocytic anemia group， the RET-He levels of the patients in mild simple microcytic anemia group and mild microcytic hypochromic anemia group were significantly decreased （P<0.01）. Compared with moderate macrocytic anemia group， the RET-He levels of the patients in moderate normocytic anemia group， moderate microcytic anemia group， and moderate microcytic hypochromic anemia group were significantly decreased （P<0.05 or P<0.01）；compared with moderate normocytic anemia group， the RET-He level of the patients in moderate microcytic hypochromic anemia group was significantly decreased （P<0.01）； compared with severe macrocytic anemia group and severe normocytic anemia group， the RET-He level of the patients in severe microcytic hypochromic anemia group was significantly decreased （P<0.01）；compared with moderate simple microcytic anemia group， the RET-He level of the patients in severe simple microcytic anemia group was significantly increased （P<0.01）； compared with mild microcytic hypochromic anemia group， the RET-He levels of the patients in moderate microcytic hypochromic anemia group and severe microcytic hypochromic anemia group were significantly decreased （P<0.01）；compared with moderate microcytic hypochromic anemia group， the RET-He level of the patients in severe microcytic hypochromic anemia was significantly decreased （P<0.01）.The cutoff value of RET-He level for diagnosing microcytic hypochromic anemia was 27.25 pg， and the sensitivity was 91.2%， the specificity was 90.5%， and the AUC was 0.968［95% confidence interval （CI）：0.952—0.984］， suggesting that the RET-He level could make an auxiliary diagnosis for microcytic hypochromic anemia. Conclusion The RET-He level can assist in the diagnosis of anemia with different morphological types and degrees， and has certain clinical application value for the diagnosis of microcytic hypochromic anemia.

Objective To discuss the effect of ganoderic acid A （GAA） on the proliferation， apoptosis， and migration ability of the PC-9 cells， and to clarify its mechanism. Methods The non-small cell lung cancer （NSCLC） PC-9 cells were cultured in vitro and divided into control group （without GAA）， low dose of GAA group （0.1 mmol·L^－1 GAA）， and high dose of GAA group （0.5 mmol·L^－1 GAA）. MTT method was used to detect the proliferation rates of the PC-9 cells in various groups； flow cytometry was used to detect the apoptotic rates of the PC-9 cells in various groups； scratch wound healing assay was used to detect the scratch healing rates of the PC-9 cells in various groups； Transwell chamber assay was used to detect the migration abilities of PC-9 cells in various groups； the expression levels of vascular endothelial growth factor （VEGF） and hypoxia-inducible factor-1α （HIF-1α） mRNA and protein in the PC9 cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. Results The MTT assay results showed that compared with control group， the proliferation rate of the cells in low dose of GAA group was decreased after treated for 48 and 72 h （P<0.05）； after treated for 24， 48， and 72 h， the proliferation rate of the cells in high dose of GAA group was decreased （P<0.05）； compared with low dose of GAA group， the proliferation rate of the cells in high dose of GAA group was decreased after treated for 24， 48， and 72 h （P<0.05）.The flow cytometry results showed that compared with control group and low dose of GAA group， the apoptotic rate of the cells in high dose of GAA group was increased （P<0.05）. The cell scratch wound healing assay results showed that compared with control group and low dose of GAA group， the scratch wound healing rate of the cells in high dose of GAA group was decreased （P<0.05）. The Transwell chamber assay results showed that compared with control group and low dose of GAA group， the number of migration cells in high dose of GAA group was decreased （P<0.05）. The RT-qPCR and Western blotting results showed that compared with control group and low dose of GAA group， the expression levels of HIF-1α and VEGF mRNA and proteins in the cells in high dose of GAA group were decreased （P<0.05）. Conclusion High dose of GAA can inhibit the proliferation of the PC-9 cells and promote the apoptosis， and its mechanism may be related to regulating the expressions of VEGF and HIF-1α mRNA and proteins.

Objective To discuss the effect of head acupuncture on the neurological function and the expressions of hypoxia-inducible factor （HIF）-1α and vascular endothelial growth factor receptor 2 （VEGFR2） in the brain tissue of rats with focal cerebral ischemia， and to clarify the related mechanism. Methods The modified middle cerebral artery occlusion（MCAO） method was used to prepare the focal cerebral ischemia rat models.Twenty successfully modeling rats were randomly divided into model group and treatment group， and there were 10 rats in each group；another 10 SD rats were selected as sham operation group.The rats in treatment group were treated with scalp acupuncture. The neurological function scores of the rats in various groups were recorded and the model was verified； Morris water maze test was used to detect the learning and memory functions of the rats in various groups； enzyme-linked immunosorbent assay （ELISA） method was used to detect the level of HIF-1α in serum and brain tissue of the rats in various groups；Western blotting method was used to detect the expression levels of HIF-1α and VEGFR2 proteins in brain tissue of the rats in various groups. Results The neurological function scores detection results showed that there was no neurological deficit of the rats in sham operation group； compared with sham operation group， the neurological function scores of the rats in model group and treatment group were increased （P<0.05）. The Morris water maze test results showed that compared with sham operation group， the time to find the platform of the rats in model group was increased （P<0.05）； compared with model group， the time to find the platform of the rats in treatment group was decreased （P<0.05）. The ELISA results showed that compared with sham operation group， the levels of HIF-1α in serum and brain tissue of the rats in model group were increased （P<0.05）；compared with model group， the levels of HIF-1α in serum and brain tissue of the rats in treatment group were increased （P<0.05）. The Western blotting results showed that compared with sham operation group， the expression levels of HIF-1α and VEGFR2 proteins in brain tissue of the rats in model group were increased （P<0.05）； compared with model group， the expression levels of HIF-1α and VEGFR2 proteins in brain tissue of the rats in treatment group were increased （P<0.05）. Conclusion The head acupuncture may restore the function of brain tissue in the rats with focal cerebral ischemia and up-regulate the levels of HIF-1α and VEGFR2 in brain tissue of the rats， and its related mechanism may be related to activating the HIF/VEGFR2 angiogenesis pathway.

Objective To discuss the effect of hydrogel-based delivery of basic fibroblast growth factor （bFGF） on the function of the NIH3T3 fibroblast cells after oxygen-glucose deprivation （OGD），and to clarify the improvement effect of bFGF-loaded hydrogel on the oxidative stress responses in the fibroblasts under the OGD condition. Methods The bFGF-containing agarose hydrogel was prepared， and the NIH3T3 cells at logarithmic growth phase were divided into blank group， 20% agarose group， and 20% agarose + glutaraldehyde group. The viabilities of the NIH3T3 cells after co-cultured with the hydrogel-eluted fluid for 6， 12， and 24 h were detected by CCK-8 assay. The NIH3T3 cells were divided into control group， OGD group， and OGD + different masses of bFGF-loaded groups （OGD + 1 ng bFGF group， OGD+10 ng bFGF group， and OGD+100 ng bFGF group）. The expression levels of α-smooth muscle actin （α-SMA）， type Ⅰ collagen， and type Ⅲ collagen proteins in the NIH3T3 cells in various groups were detected by Western blotting method； the content of malondialdehyde （MDA） and activities of lactate dehydrogenase （LDH） and superoxide dismutase （SOD） in the NIH3T3 cells were detected by assay kits； the in vitro release rate of bFGF in the NIH3T3 cells in various groups was detected by enzyme-linked immunosorbent assay （ELISA） method. Results The CCK-8 assay results showed that compared with blank group， the viability of the NIH3T3 cells between blank group， 20% agarose group， and 20% agarose+ glutaraldehyde group at different elution times had no significant difference（P>0.05）. The Western blotting results showed that the expression levels of α-SMA and type Ⅰ collagen proteins in the NIH3T3 cells in OGD group were significantly higher than that in control group （P<0.05 or P<0.01）.Compared with OGD group，the expression levels of α-SMA and type Ⅰ collagen proteins in the NIH3T3 cells in OGD+1 ng bFGF， OGD+10 ng bFGF， and OGD+100 ng bFGF groups were significantly decreased （P<0.05 or P<0.01） in a dose-dependent manner.Compared with control group，the level of MDA and activity of LDH in the NIH3T3 cells in OGD group were increased（P<0.05）， while the activity of SOD was decreased （P<0.05）. Compared with OGD group， the level of MDA and the activity of LDH in the NIH3T3 cells in OGD + 1 ng bFGF group were decreased （P<0.05）， while the activity of SOD was increased （P<0.05）.The ELISA results showed that about 10% bFGF was released from the hydrogel within 4 h， about 15% bFGF was relleased from the bydrogel with in 12 h， and about 21% bFGF was released from the hydrogel within 24 h. Conclusion The bFGF-loaded hydrogel can modulate the transformation of the fibroblasts under the OGD condition，inhibit the expression of typeⅠ collagen protein，and improve the oxidative stress responses in the fibroblasts. Moreover，this system exhibits a sustained drug-release ability.

Objective To discuss the influence factors of retromolar space in the patients with different osteofacial types， and to analyze the correlation between the mandibular bone structure and retromolar space and craniofacial structure. Methods A total of 200 adult orthodontic patients with fully erupted dentition except the third molars and crowding less than 5 mm were chosen and divided into skeletal type Ⅰ low angle group， skeletal type Ⅰ average angle group， skeletal type Ⅰ high angle group， skeletal type Ⅱ average angle group， and skeletal type Ⅲ average angle group. The sella nasion A point angle（SNA）， sella nasion B point angle（SNB）， AB plane angle（ANB）， skull base anterior-mandibular plane angle（SN-MP），facial height index（FHI），mandibular ramus length（Go-Co），and mandibular body length（Go-Gn） of all the patients were detected. Cone-beam computed tomography （CBCT） technology was used to detect the shortest distances C0 and C2 from the mandibular second molar to the anterior border of the ramus at the planes parallel to the cuspid line with a depth of 0 and 2 mm form the occlusal plane， and the shortest distances R2， R4， R6， R8， and R10 from the tooth root to the lingual cortical bone of the mandible at the enamel-bone junction in the root direction 2， 4， 6， 8， and 10 mm planes.The correlation was analyzed by using Spearman correlation analysis. Results Compared with skeletal type Ⅰ low angle group， the SN-MP of the patients in skeletal type Ⅰ average angle group and skeletal type Ⅰ high angle group was increased significantly （P<0.01）， and the FHI was decreased significantly （P<0.01）. Compared with skeletal type Ⅰ average angle group， the SN-MP of the patients in skeletal type I high angle group was increased significantly （P<0.01），FHI was decreased significantly （P<0.01）. Compared with skeletal type Ⅰ average angle group， the ANB of patients in skeletal type Ⅱaverage angle group was significantly increased （P<0.01）， while the ANB of the patients in skeletal type Ⅲ average angle group were significantly decreased （P<0.01）. Compared with skeletal type Ⅱ average angle group， the ANB of the patients in skeletal type Ⅲ average angle group was significantly decreased （P<0.01）. Under the C2， R2， R4， R6， R8， R10 planes and in the Go-Co and Go-Gn， compared with skeletal type Ⅰ low angle group， the retromolar spaces of the patients in skeletal type Ⅰ average angle group and the skeletal type Ⅰ high angle group were significantly decreased （P<0.01）； compared with skeletal type Ⅰ average angle group， the retromolar space of the patients in skeletal type Ⅰ high angle group was significantly decreased （P<0.01）. Under the C0 plane， compared with skeletal type Ⅰ low angle group and the skeletal type Ⅰ average angle group， the retromolar space of the patients in skeletal type Ⅰ high angle group was decreased （P<0.01）. Under the C0， R2， R4， R6， R8， and R10 planes， compared with skeletal type Ⅰ average angle group， the retromolar space of the patients in skeletal type Ⅱ average angle group was significantly decreased （P<0.01）， the retromolar space of the patients in skeletal type Ⅲ average angle group was increased （P<0.01）； compared with skeletal type Ⅱ average angle group， the retromolar space of the patients in skeletal type Ⅲ average angle group was increased （P<0.01）.Under the C2 plane and in the Go-Co and Go-Gn， compared with skeletal type Ⅰ average angle group and skeletal type Ⅱ average angle group， the retromolar space of the patients in skeletal type Ⅲ average angle group was increased （P<0.01）. The retromolar space，Go-Co and Go-Gn of the patients in skeletal type Ⅰ low angle group， skeletal type Ⅰ average angle group， and skeletal type Ⅰ high angle group under all planes had positive correlations with the SNB and FHI （P<0.01）， and a negative correlation with SN-MP （P<0.01）；the retromolar space at all levels had a positive correlation with Go-Co （P<0.01）； except for the C2 plane， the remaining retromolar space under all planes had positive correlations with Go-Gn （P<0.01）；there was a positive correlation between Go-Co and Go-Gn （P<0.01）；there was a negative correlation between Go-Gn and ANB（（P<0.01）.The retromolar space and Go-Co and Go-Gn of the patients in skeletal type Ⅰ average angle group， skeletal type Ⅱ average angle group， and skeletal type Ⅲ average angle group under all planes had positive correlations with SNB （P<0.01），and the retromolar space had positive correlations with Go-Co and Go-Gn （P<0.01）； there was a positive correlation between Go-Co and Go-Gn （P<0.01）；there was a positive correlation between Go-Co and FHI（P<0.01）. Conclusion The skeletal type is an important influence factor of the retromolar space of mandible， when the molar distalization plan is formulated， the skeletal type of the patient should be considered； CBCT should be used to evaluate the available space for mandibular molar distalization.

Objective To discuss the effect factors of enteral nutrition feeding intolerance （FI） of the patients with severe acute pancreatitis （SAP）， and to construct and verify the risk prediction model. Methods The data of 246 SAP patients were collected and the patients were divided into tolerance group （n=143） and non-tolerance group （n=103） according to whether the enteral nutrition FI occurred.Age， intra-abdominal pressure， hypertriglyceridemia， hypoproteinemia， and other related data of the patients in two groups were compared， the predictive factors of FI were screened out by Logistic regression analysis，and the prediction model was constructed.The area under receiver operating characteristic （ROC） curve （AUC） and Hosmer-Lemeshow test were used to verify the prediction effect of the model. Ninety-two SAP patients were selected to verify the predictive effect of the model. Results A total of 6 predictors，such as hypertriglyceridemia （OR=1.962， 95%CI：1.088－3.538，P=0.025），hypoproteinemia（OR=1.920， 95%CI：1.049－3.516，P=0.034），acute physiological and chronic health status Ⅱ（APACHEⅡ）≥score 20 points （OR=3.118， 95%CI：1.720－5.653，P<0.01），intra-abdominal pressure ≥12 mmHg （OR=2.826， 95%CI：1.534－5.205，P=0.001），time to start enteral nutrition ≥72 h（OR=2.350，95%CI：1.179－4.683，P=0.015）， and addition of microecological agent （OR=0.379， 95%CI：0.209－0.685，P=0.001） were included. The formula of the model was Z=－1.206+0.674×hypertriglyceridemia+0.652×number of patients with hypoproteinemia+1.137×number of patients with APACHEⅡ score ≥20 points +1.039×number of patients with intra-abdominal pressure ≥12 mmHg+0.855×number of patients with time to start enteral nutrition ≥72 h－0.971×number of patients with　addition of microecological agent. The Hosmer-Lemeshow test results showed that χ²=5.985 and P=0.901， and the AUC of the prediction model was 0.793 （95%CI： 0.735－0.851， P<0.01），the Yoden index was 0.498，the optimal critical value was 0.499， the sensitivity was 0.818， and the specificity was 0.680. The agent verification results of the model showed that the AUC was 0.880 （95%CI： 0.811－0.948，P<0.01），the sensitivity was 0.774，the specificity was 0.867， and the accuracy rate was 81.52%. Conclusion The constructed risk prediction model of SAP enteral nutrition FI has high sensitivity and specificity， and good prediction efficiency.

Objective To analyze the clinical characteristics and risk factors of the patients with primary Sjogren’s syndrome （pSS） complicated with interstitial lung disease （ILD）， and to improve the clinicians’understandings of the disease， and to provide the evidence for the early detection of extradular organ involvement by the clinicians. Methods The clinical characteristics of 176 patients with pSS were collected and were divided into pSS complicated with ILD group （n=21） and pSS complicated without ILD group （n=155） according to whether the patients were complicated with ILD.The levels of hemoglobin （Hb）， white blood cells （WBC）， platelets （PLT）， C-reactive protein （CRP）， erythrocyte sedimentation rate （ESR），rheumatoid factor （RF），immunoglobulin G （IgG），immunoglobulin M （IgM），immunoglobulin A （IgA）， complement 3 （C3）， complement 4 （C4）， and the positive rates of granular type antinuclear antibodies， cytoplasmic granular type antinuclear antibodies， homogeneous type antinuclear antibodies， anti-SSA， anti-SSB， and anti-Ro-52 antibodies of the patients in two groups were compared.Univariate and multivariate Logistic regression analysis were used to analyze the clinical characteristics and laboratory indexes of the pSS patients complicated with ILD. Results Among the 176 pSS patients， the ratio of male to female was 1∶28， with the median age of 49 （39－58） years and the median course of 24 （24－48） months. Compared with pSS complicated without ILD group，the age of the patients in ILD group was older （P<0.01）， and the level of ESR was increased（P<0.01），the positive rates of cytoplasmic granular type antinuclear antibody and anti-Ro-52 antibody were increased （P<0.01）. The multivariate Logistic regression analysis results showed that the increasing of ESR level （OR=1.046，95% CI： 1.026－1.066， P=0.001） and positive granulosa type （OR=4.676，95% CI： 1.156－18.916， P=0.031） were the risk factors for ILD complicated with pSS. Conclusion The patients with pSS complicated by ILD have an older onset age and higher ESR levels； ILD is more likely to occur when the cytoplasmic granular type antinuclear antibody and anti-Ro-52 antibody of the patients are positive； the increasing of the ESR level and positive coated granular type antinuclear antibody are the risk factors for the pSS complicated with ILD.

Objective To analyze the clinical data of the patients with non-syndromic supernumerary teeth （NSMST）， and to explore the possible mechanism of occurrence of supernumerary teeth， diagnostic criteria，and treatment method， and to enhance the comprehension of the clinicians about this disease. Methods The clinical data of one patient with NSMST were collected. Based on the specialist examination and relevant literature review results， the causes， diagnostic points and treatment principles of the supernumerary teeth were analysed. Results The patient， a 19-year-old male， checked in the clinic due to his subjective feeling of facial asymmetry， denying any systemic disease or history of genetic disorders. The intraoral examination results showed 4 supernumerary teeth， all of them showed the resembling molars morphology. The cone beam CT （CBCT） results showed that the roots of 4 erupted supernumerary teeth were fully developed. One impacted supernumerary tooth could be seen on the palatal root side of tooth 18， resembling a premolar， with fully developed roots， and there was a calcified crown shadow being observed distal mesially， and no root formation could be seen. The X-ray cephalometric analysis results showed a smaller SNB angle than the normal value， indicating the deficiency in the mandibular development； a larger FH-MP angle indicated a steep mandibular plane and high-angle pattern. There were a total of 6 supernumerary teeth on the right maxillary molar area， so the diagnosis was NSMST. The surgical extraction of the supernumerary teeth was recommended， followed by combined orthognathic-orthodontic treatment to restore normal facial morphology and dental arch form. The patient did not undergo treatment due to his personal reasons. Conclusion The NSMST patients needs to be definitely diagnosed through the imaging examinations clinically，the surgical removal is the principle treatment method，and early discovery and treatment can minimise the impact on the patient’s facial region and dentition.

Objective To analyze the imaging performance and clinical diagnosis and treatment process of one patient with peri-gallbladder fluid caused by the posterior duodenal perforation primarily diagnosed by ultrasound， and to provide the clinical diagnostic evidence for this disease. Methods The clinical data， laboratory examination， gastroscope performance， and imaging performance of one patient with peri-gallbladder fluid caused by the posterior duodenal perforation was collected. The process of diagnosis and treatment was recorded and followed up. The related literatures were reviewed to analyze the clinical characteristics and imaging performances of the posterior duodenal perforation. Results The patient was a 50-year-old male who had constant dull pain in the upper right abdomen for over 20 d， without obvious cause and intensified after meals， accompanied by radiculalgia in the right waist and back.The ultrasonic examination at a local hospital showed an occupying lesion in the gallbladder， and the patient came to our hospital for the further diagnosis and treatment. On the day of admission， the ultrasound examination showed poor fasting gallbladder filling， continuous and uniformly thickened gallbladder wall.The multiple strong echoes were observed in the gallbladder cavity， and the chaotic distributed hypoechoic and anechoic areas could be seen around the gallbladder， extending to the back of the duodenal bulb， and connected with the bulb by a narrow strip of gaseous shadow. The ultrasound results showed that there was perforation of the posterior wall of the duodenal bulb， leading to pericholecystic fluid accumulation， with poor gallbladder filling due to compression， and chronic inflammation of the gallbladder wall associated with gallstones. The enhanced computed tomography （CT） results clearly showed the perforation site in the posterior wall of the duodenal bulb， and the CT conclusion was consistent with the ultrasound conclusion. The gastroscope results showed a large deep ulcer on the posterior wall of the duodenal bulb， and it confirmed to be diagnosed as the peri-gallbladder fluid caused by ulcer perforation. A gastric tube and jejunal nutrition tube were inserted， and the symptomatic treatments such as anti-inflammation， analgesia， and acid suppression were carried out. The ultrasound reexamination within 3 months of treatment showed the peri-gallbladder fluid was gradually decreased and the gallbladder cavity was gradually filled.Since the patient’s gallbladder inflammation symptoms persisted，the cholecystectomy was performed， and it was pathologically diagnosed as chronic cholecystitis complicated with gallstones. Conclusion The posterior duodenal perforation is often misdiagnosed or missed due to atypical clinical manifestations.The imaging examination can assist in early clinical diagnosis. When the presence of abdominal gas does not interfere with observation， the ultrasound can serve as the first choice and effective method for the diagnosis of gastrointestinal perforation.

Objective To discuss the risk factors of recurrence of papillary thyroid microcarcinoma （PTMC），and to provide the reference for the individualized diagnosis and treatment of PTMC. Methods The retrieval time was limited from the establishment to April 2022， and the case-control studies related to PTMC recurrence searched in PubMed， EMBase， Elsevier Science Direct and Web of Science Databases were collected.Review Manager 5.4 software was used to perform the Meta-analysis and ADDIS 1.16.8 software was used to make the Bayesian network Meta-analysis，and the risk factors of PTMC recurrence were screened out and ranked. Results Fourteen studies were included， involving 7 834 PTMC patients who underwent surgical treatment，302 patients experienced lymphnode recurrence， local thyroid bed recurrence， or distant metastasis recurrence. Age， multifocality， tumor size， extrathyroidal invasion， lymphnode metastasis， and vascular invasion of the patients were related to the PTMC recurrence （P<0.05）. Lymphnode metastasis was the most significant risk factor for the PTMC recurrence， followed by vascular invasion， and multifocality was the least significant risk factor. There were no significant correlations between gender， lesion histopathological background， gland histopathological background， radioactive iodine treatment， surgical margin， and the extent of thyroidectomy with PTMC recurrence （P>0.05）. Conclusion Lymphnode metastasis， vascular invasion， tumor size>5 mm，age，extrathyroidal invasion， and multifocality are the risk factors for the PTMC recurrence.The clinician may consider to treat the PTMC patients with routine central lymphnode dissection to prevent disease recurrence，and develop the individualized treatment plans based on the risk factor stratification.

Objective To discuss the method of primary culture and activity evaluation system of the neural stem cells （NSCs） of the fetal rats， and to confirm the optimal subculture conditions of the NSCs. Methods The SD rats with 11－14 d gestation were chosen，and the primary NSCs of the fetal rats were extracted. Nestin immunofluorescence staining was used to identify the NSCs. The NSCs were subcultured at the cell densities of 1.0×10⁴，2.0×10⁴，6.0×10⁴，1.0×10⁵， 1.6×10⁵ and 2.0×10⁵ mL^－1， then the morpholgy of the neurospheres was observed 48 h after passage and the diameter of the neurospheres was calculated. The survival rates of NSCs in the neurospheres with different diameters were detected by live-dead cell staining. Results The expression of nestin in the NSCs was positive， indicating that the cultured cells were the NSCs. The NSCs showed aggregation growth and strong cell-cell adhesion pattern forming neurospheres with an average diameter of （152.72±47.52） μm， and there were few single cells scattered between the neurospheres. Compared with 2.0×10⁴ mL^－1 subculture density，the diameters of the neurospheres at the subculture densities of differences 6.0×10⁴ mL^－1 and 1.0×10⁵ mL^－1 were increased （P<0.05）. There were no significant differences in the diameters of the neurospheres at the subculture densities of 1.0×10⁵ mL^－1， 1.6×10⁵ mL^－1， and 2.0×10⁵ mL^－1 （P>0.05）. Compared with neurospheres with diameters of 0—40 μm， 40—60 μm， 60—80 μm，and 80—100 μm， the survival rate of NSCs in the neurospheres with the diameters of 100—200 μm was decreased （P<0.05）. Conclusion The neurospheres with the diameters of 80—100 μm when subcultured at the density of 1.0×10⁵ mL^－1 can effectively improve the efficiency and activity of subculture of the NSCs.

Objective To improve the pre-processing laboratory test method for the severe acute respiratory syndome，coronavirus （SARS-CoV-2） specimens， and to evaluate its effectiveness. Methods A total of 90 SARS-CoV-2 negative specimens and 30 SARS-CoV-2 positive specimens were collected. The specimens were divided into traditional group， modified 1 group， and modified 2 group， according to the test method. The specimens in traditional group were inactivated by using the guanidine salts and a 56 ℃， 30 minute water bath， followed by single tube oscillation， specimen de-identification， and single-handed lid opening for testing. The specimens in modified 1 group were inactivated by using the guanidine salts， followed by single tube oscillation， specimen de-identification， and single-handed lid opening for testing. The specimens in modified 2 group were inactivated by using the guanidine salts， followed by batch oscillation in vertical， horizontal， and 180-degree angle， specimen de-identification， and single-handed lid opening for testing. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the cycle threshold （Ct） values and detection time of the specimens in various groups； Spearman correlation analysis was used to analyze the correlation of Ct values between various groups； Blank-Altman method was used to perform the consistency test； the detection time of SARS-CoV-2 of the specimens in various groups was recorded. Results The Ct values of the specimens in modified 1 group were strongly correlated with those in traditional group （P<0.01）， and the correlation coefficient（r）=0.975 4 （0.966 2－0.982 1）. The Ct values of the specimens in modified 2 group were strongly correlated with those in traditional group （P<0.01）， r=0.986 2 （0.980 9－0.990 0）. The inter-group deviation of detecting SARS-CoV-2 negative specimens between modified 1 group and traditional group was 0.158 2（－0.613 7－0.930 1）， and the inter-group deviation of detecting SARS-CoV-2 positive specimens was 0.117 0 （－0.403 1－0.637 1）， indicating good consistency.The inter-group deviation between modified 2 group and traditional group for detecting SARS-CoV-2 negative specimens was 0.015 7（－0.550 4－0.581 8）， and for detecting SARS-CoV-2 positive specimens was 0.022 7（－0.454 1－0.499 4）， which indicated the good consistency. The detection time of 90 SARS-CoV-2 specimens in modified 1 group was 15 min， while the detection time of 90 SARS-CoV-2 specimens in modified 2 group was 10 min and the detection time of 90 SARS-CoV-2 specimens in traditional group was 45 min. Compared with traditional group， the detection time in modified 1 group was reduced by 67%and the detection time in modified 2 group was reduced by 11% . Conclusion The modified SARS-CoV-2 specimen testing method shows strong correlation and good consistency with the traditional testing method. It can save the detection time and improve the efficiency.

Objective To discuss the resistance and regeneration effects of long non-coding RNA （lncRNA） GPRC5D-AS1 on the muscle atrophy of myocytes in the mice induced by dexamethasone， and to clarify its mechanism. Methods The C2C12 cells at logarithmic growth phase were selected. The survival rates of the cells after treated with 0， 25， 50， 75， 100， 200 and 400 mg·L^－1 dexamethasone for 24， 48 and 72 h were detected， which was used to select the optimal dose and time to induce the muscle atrophy model.The expression level of lncRNA GPRC5D-AS1 in the C2C12 cells was detected by real-time fluroscence quantitative PCR（RT-qPCR） method to validate the cell model. The C2C12 cells were divided into normal group， control group，lncRNA GPRC5D-AS1-NC group，and lncRNA GPRC5D-AS1-OE group. The expression levels of lncRNA GPRC5D-AS1 in the cells in various groups were detected by RT-qPCR method. The levels of reactive oxygen species （ROS）， glutathione peroxidase 4 （GPX4）， ferrous ion （Fe²⁺） and malondialdehyde （MDA） in the cells in various groups were detected by flow cytometry and enzyme-linked immunosorbent assay （ELISA） method； the ultrastructural changes of mitochondria in the cells in various groups were observed under transmission electron microscope； the expression of myogenic differentiation protein （MyoD） in the cells in various groups was detected by immunofluorescence； the expression levels of ferroptosis-related proteins in the cells in various groups were detected by Western blotting method. Results The best modeling effect was achieved with 100 mg·L^－1 dexamethasone at 48 h. Compared with normal C2C12 cells， the expression level of lncRNA GPRC5D-AS1 in the C2C12 cells after treated with 100 mg·L^－1 dexamethasone for 48 h was decreased （P<0.05）， confirming that lncRNA GPRC5D-AS1 had a regulatory role in the muscle atrophy model. The RT-qPCR results showed that compared with normal group， the expression level of lncRNA GRPC5D-AS1 in the cells in model group was decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression level of lncRNA GRPC5D-AS1 in the cells in lncRNA GRPC5D-AS1-OE group was increased （P<0.05）. The flow cytometry results showd that compared with normal group， the ROS level in the cells in model group was increased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the ROS level in the cells in lncRNA GRPC5D-AS1-OE group was decreased （P<0.05）. The ELISA results showed that compared with normal group， the levels of Fe²⁺ and MDA in the cells in model group were increased （P<0.05）， and the level of GPX4 was decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the levels of Fe²⁺ and MDA in the cells in lncRNA GRPC5D-AS1-OE group were decreased （P<0.05）， and the level of GPX4 was increased （P<0.05）. The transmission electron microscope results showed that the cells in normal group were round and normal in size with rich cilia on the membrane， and the mitochondria had a normal morphology； the cells in model group had less mitochondria and granulation in the cytoplasm， with blurred and swollen mitochondrial structures and typical ferroptosis-related manifestations； the number of mitochondria in the cells in lncRNA GRPC5D-AS1-OE group was increased，and the granulation in cytoplasm of the cells was decreased， and showed clearer mitochondrial structures. The immunofluorescence detection results showed that compared with normal group，the expression of MyoD in the cells in model group was decreased and the cells exhibited muscle atrophy； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression of MyoD in the cells in lncRNA GRPC5D-AS1-OE group was increased， and the cells exhibited less muscle atrophy and more regeneration of myocytes. The Western blotting results revealed that compared with the normal group， the expression level of long-chain acyl-coenzyme A synthase （ACSL）4 protein in the cell in model group was increased （P<0.05）， and the expression levels of solute carrier family 7 member 11 （SLC7A11） and GPX4 proteins were decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression level of ACSL4 protein in the cells in lncRNA GRPC5D-AS1-OE group was decreased （P<0.05）， and the expression levels of SLC7A11 and GPX4 proteins were increased （P<0.05）. Conclusion LncRNA GPRC5D-AS1 can protect the ferroptosis in the myocytes induced by dexamethasone and promote the repairment and regeneration of the cells，and its mechanism is related to regulating the SLC7A11/GPX4/ACSL4 signaling pathway.

Objective To discuss the risk factors of renal function preservation and trifecta of the patients with renal cell carcinoma（RCC） after robot-assisted partial nephrectomy （RAPN）， and to provide the evidences for the preperative evaluation，postoperative treatment and long-term follow-up. Methods A retrospective analysis was conducted on the the clinical data of 111 cases of RCC patients undergoing RAPN.The patients were divided into trifecta group （n=73） and non-trifecta group（n=38） according to whether the trifecta outcome was achieved； according to the changes of 24 h etimated glomerular fltration rates（eGFR） before and after operation，the patients were divided into postoperative 24 h eGFR decreasing≤10% group（n=85） and postoperative 24 h eGFR decreasing>10% group（n=26）.The age， gender，American Society of Anesthesiologists（ASA） score， body mass index （BMI）， hypertension，diabetes，preoperative eGFR，percentages of postoperative 24 h eGFR change， hilar tumor，dorsal ventral positions of tumor，maximum tumor diameters， sugrical aproaches，warm ischemia time（WIT），intraoperative estimated blood loss （EBL），tumor pathological types，tumor TNM stages， RENAL scores， PADUA scores， central indexes （C-index）， renal tumor invasion indexes （RTII），and tumor contact area （CSA）of the patients in two groups were compared.Multivariate Logistic regression was used to analyze the influencing factors of the patients’ achievement of trifecta and the postoperative 24 h eGFR decreasing>10%. Multiple linear regression was used to analyze the influencing factors of the changes of postoperative 24 h eGFR. Results A total of 73 patients achieved trifecta among all the 111 patients.The univariate analysis results showed that there were statistically significant differences in age， hypertension， tumor maximum diameter， RENAL score， PADUA score， C-index， RTII， CSA，and EBL of the patients in trifecta group and non-trifecta group. The multivariate Logistic analysis results showed that EBL was an independent influencing factor for failing to achieve trifecta of the patients after RAPN（OR=1.006，95%CI=1.001—1.011，P=0.020）.There were statistically significant differences in tumor maximum diameters， RENAL scores， PADUA scores， C-indexes， RTII， CSA， WIT， EBL， and tumor TNM stages of the patients（P<0.05） in 24 h postoperative eGFR decreasing > 10% and postoperative 24 h eGFR decreaing ≤10% groups. The multivariate Logisitc regression analysis results showed that RTII was an independent influencing factor for the postoperative 24 h eGFR decreasing> 10% of the patients （OR=4.442，95%CI=1.049—18.806，P=0.043）.The tumor maximum diameter，RENAL，PADUA，C-index， RTII， CSA， WIT， EBL，and tumor TNM stage had no correlation with the postoperative eGFR changes； RTII had negative correlationship with postoperative 24 h eGFR changes（B=－7.204，95%CI=－14.305－－0.102，P=0.047）. Conclusion EBL is an independent influencing factor for failure to achieve trifecta outcomes of the patients after RAPN； RTII is negatively correlated with the postoperative 24 h eGFR changes.

objective To discuss the morphological changes and related factors of soft and bone tissues in chin in the skeletal class Ⅲ malocclusion patients at three dimensions after orthognathic operation. Methods A total of 19 adult patients underwent orthognathic operation were collected， including 9 males and 10 females. All the patients received craniofacial computed tomography （CT） scans one week before operation（T0） and 12 months after （T1） operation to obtain the facial data of the soft and bone tissues， with a three-dimensional reconstruction and registration by using ProPlan CMF 3.0 software. The measurements and calculations were made on the thickness of chin soft tissue， the horizontal， sagittal， and vertical changes in anatomical landmarks， and the correlation and ratios between them. Results The most significant retrusion of soft tissue after operation was in the lower lip vermilion and the mentolabial fold area. The posterior displacement of soft tissue was gradually decreased， spreading from this area to the periphery. The posterior change in the submental area was relatively minor， with an anterior trend at the submental point. Compared with T0 time point， the soft tissue thickness at the lower lip （LL）at T1 time point was increased （P<0.05）.The linear regression analysis results showed that at the horizontal（X）， as the bone tissue changed to the left，the anatomical landmarks of soft tissue in chin LL （B=0.795， R2=0.832）， mentolabial sulcus （Si） （B=0.876，R2=0.987）， soft tissue pogonion （Pos） （B=0.890， R2=0.971）， and menton of soft tissue（Mes） （B=0.942，R2=0.978） showed negative changes， all moved to the left； the movement directions of soft and bone tissues were consistent. At the sagittal （Y） direction， as the bone tissue moved backward， the LL （B=0.882， R2=0.934）， Si （B=0.946， R2=0.847）， Pos （B=0.839， R2=0.909） and Mes （B=0.666，R2=0.455） all moved backward，and the movement directions of soft and bone tissues were consistent.At the vertical （Z） direction， as the bone tissue moved upwards， the LL （B=0.932， R2=0.686）， Si （B=0.834， R2=0.469）， and Mes （B=0.925， R2=0.709） moved downwards， while the Pos （B=0.487， R2=0.444） moved upwards. In horizontal， sagittal， and vertical directions， the correlation between soft-tissue movement（ΔSM） and bone-tissue movement（ΔBM） of chin anatomical landmarks was generally strong （0.6<r<1.0，P<0.01）. Conclusion The changes of soft and hard tissues in the chin of skeletal class Ⅲ malocclusion patients at horizontal， sagittal， and vertical directions after orthognathic operation are influenced by bone tissue movements， showing the positive correlation. The correlations at the horizontal and sagittal directions are stronger， and the correlation at the vertical direction is weaker.

Objective To discuss the effect of shear wave elastography （SWE） on the differential diagnosis of benign and malignant breast non-mass lesion （NML）， and to clarify its clinical significance. Methods A total of 158 patients with breast NML were selected and divided into benign lesion group （ n=83） and malignant lesion group（ n=75） according to the postoperative pathological type of the patients.The routine ultrasound and SWE examinations were conducted before operation， and the SWE parameters of the lesions， including the maximum （E_max）， average （E_mean），minimum （E_min）， standard deviation （E_sd） and whether the stiff rim sign presented or not at 1， 2 and 3 mm around the lesion were recorded. The SWE characteristics and differences in SWE parameters of the NML patients between two groups were analyzed， and the receiver operating characteristic curve（ROC） was plotted， and the area under curve （AUC）， specificity and sensitivity were calculated. Results There were statistically significant differences in age， lesion size， palpation， and whether menopause had crested of the NML patients between two groups（ P<0.01）. All the lesions of the patients in two groups appeared as hyperechoic and irregular in shape. Compared with benign lesion group， the percentage of the patients with non-parallel， calcification and posterior echo attenuation in malignant lesion group was significantly increased （ P<0.01）， and the percentage of the patients without blood supply was significantly decreased （ P<0.01）. Compared with benign lesion group， the E_max， E_mean， and E_sd of the NML patients in malignant lesion group were significantly increased （ P<0.01）， and the E_max， E_mean， and E_sd at 1， 2， and 3 mm around the lesion were all increased （ P<0.01）. The stiff rim sign manifested as a circular or semi-circular red region around the lesions of the patients in malignant lesion group. Compared with benign lesion group， the percentage of hard rim sign of the NML patients in malignant lesion group was significantly increased （ P<0.01）.Compared with inside the lesion，the E_min at 1，2 and 3 mm around the lesion of the NML patients in benign lesion group and malignant lesion group were decreased（ P<0.01）. The AUC of E_max at 1， 2 and 3 mm around the lesion were 0.872， 0.860， 0.873 and 0.866，respectively； the AUC of E_mean were 0.796， 0.822， 0.820 and 0.815，respectively； the AUC of E_sd were 0.832， 0.857， 0.859， and 0.842，respectively；the sensitivity and specificity of E_max inside the lesion were 85.33% and 83.13%，respectively；the sensitivity and specificity of E_max at 1 mm around the lesion were 82.67% and 80.72%，respectively； the sensitivity and specificity at 2 mm around the lesion were 78.67% and 87.95%，respectively； the sensitivity and specificity at 3 mm around the lesion were 80.00% and 86.75%，respectively； the specificity of E_sd at 2 mm around the lesion was highest （93.33%）. Conclusion The SWE parameters inside and around the lesion and the manifestation of stiff rim sign can provide the reference for the differential diagnosis of benign and malignant breast NML， and have good diagnostic performance.

Objective To discuss the expression levels of cyclooxygenase-2 （COX-2） and β-catenin in the in situ endometrium tissue， lesion tissue， and serum of the adenomyosis （AM） patients，and to clarify their relationships with dysmenorrhea. Methods A total of 90 cases of confirmed AM patients with dysmenorrhea who underwent abdominal hysterectomy were selected as AM group， and 30 cases of patients with uterine myomas （UM） without dysmenorrhea were selected as UM group. The degree of dysmenorrhea of the AM patients was assessed by using the visual analogue scale （VAS）， and 90 cases of AM patients were divided into mild dysmenorrhea group， moderate dysmenorrhea group， and severe dysmenorrhea group （n=30）. Immunohistochemistry （IHC） was used to detect the expression levels of COX-2 and β-catenin proteins in endometrium tissue of the patients in two groups； enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of COX-2 and β-catenin in serum of the patients in two groups；the association between the expression levels of COX-2 and β-catenin in endometrium tissue of the patients in two groups and degree of dysmenorrhea of the patients was analyzed by Spearman correlation analysis. Results The COX-2 protein expression was mainly located in the cytoplasm of endometrium tissue， while β-catenin protein was mainly expressed in the cell membrane or cytoplasm of endometrium tissue. Compared with UM group， the expression levels of COX-2 protein in in situ endometrium tissue and lesion tissue of the patients in AM group were increased（P<0.05），and the expression level of β-catenin protein was decreased（P<0.05）. Compared with in situ endometrium tissue of the patients in AM group， the expression level of COX-2 protein in lesion endometrium tissue of the patients in AM group was increased（P<0.05）.The expression level of COX-2 protein in the lesion tissue of the AM patients with moderate dysmenorrhea and the expression level of COX-2 protein in in situ endometrium and lesion tissues of the AM patients in severe dysmenorrhea group were significantly higher than those in mild dysmenorrhea group（P<0.05）； the expression level of COX-2 protein in in situ endometrium and lesion tissues of the AM patients in severe dysmenorrhea group were significantly higher than those in moderate dysmenorrhea group（P<0.05）.There were no significant differences in the expression levels of β-catenin protein in in situ endometrium tissue and lesion tissue of the AM patients between different degrees of dysmenorrhea groups （P>0.05）. Compared with UM group，the serum level of COX-2 of the patients in AM group was significantly increased （P<0.05），and the serum levels of β-catenin of the patients in AM group was decreased （P<0.05）.Compared with mild dysmenorrhea group，the serum COX-2 levels of the patients in moderate and severe dysmenorrhea groups were increased（P<0.05）. There were no significant differences in serum β-catenin level of the patients between different degrees of dysmenorrhea groups （P>0.05）.The expression level of COX-2 protein in lesion tissue of the AM patients was negatively correlated with the expression level of β-catenin protein （r=－0.364， P<0.05）； in in situ endometrium and lesion tissues of the AM patients， the expression level of COX-2 protein was positively correlated with the degree of dysmenorrhea （r=0.511，P<0.05； r=0.696， P<0.05）， suggesting that the expression level of COX-2 protein was positively correlated with the degree of dysmenorrhea and there was no correlation between the expression level of β-catenin protein and the degree of dysmenorrhea （P>0.05）. The correlation between the serum levels of COX-2 and β-catenin of the patients were negative （r=－0.534， P<0.05）， and the COX-2 level of the patients was positively correlated with the degree of dysmenorrhea （r=0.613， P<0.05）. Conclusion The increasing of expression level of COX-2 and decreasing of expression level of β-catenin and their correlation both induce AM development and endometriotic invasion， and high expression of COX-2 may promote the dysmenorrhea， which may serve as a potential molecular target for the treatment of AM and associated dysmenorrhea.

Objective To disuss the effect of glucagon like peptide-1 receptor （GLP-1R） agonist Exendin-4（E4） on injury of cardiomyocytes in the hyperglycemia and hyperlipidemia（HGHL） environment， and to clarify its related mechanism. Methods The H9C2 cardiomyocytes of the rats were divided into control group， HGHL group， HGHL+E4 group， HGHL+E4+iron death agonist（Erastin） group.The HGHL model of the H9C2 cells was induced by 33 mmol·L^－1 glucose and 500 μmol·L^－1 palmitate，and the cells in HGHL+E4 group and HGHL+E4+Erastin group were treated with 100 nmol·L^－1 E4 and 5 μmol·L^－1 Erastin. CCK-8 assay was used to detect the survival rates of cells in various groups； Hoechst 33258 fluorescence staining was used to observe the apoptosis of cells in various groups； flow cytometry was used to detected the apoptotic rates of cells in various groups；2'，7'-dichlorodihydrofluorescein diacetate（DCFH-DA） fluorescence probe was used to detect the levels of reactive oxygen species （ROS） in the cells in various groups； the corresponding kit was used to detect the levels of glutathione （GSH） and malondialdehyde （MDA） in cells in various groups；JC-1 staining was used to detect the mitochondrial membrane potential （MMP） in cells in various groups； FerroOrange fluorescence probe was used to detect the levels of Fe²⁺ in the cells in various groups； Western blotting method was used to detect the expression levels of glutathione peroxidase 4 （GPX4） and solute carrier family 7 member 11（SLC7A11） in the cells in various groups. Results Compared with control group， the survival rate of cells in HGHL group was decreased （P<0.05），and the cells were fragmented and collapsed， which showed apoptotic morphology；the apoptotic rate and the levels of ROS and MDA in the cells were increased（P<0.05）， the levels of GSH and MMP were decreased（P<0.05）， the level of Fe²⁺ was increased（P<0.05），and the expression levels of GPX4 and SLC7A11 proteins were decreased（P<0.05）.Compared with HGHL group， the survival rate of the cells in HGHL+E4 group was increased （P<0.05），and the apoptosis was decreased；the apoptotic rate and the levels of ROS and MDA were decreased（P<0.05）， while the levels of GSH and MMP were increased（P<0.05）， the level of Fe²⁺ was decreased（P<0.05）， and the expression levels of GPX4 and SLC7A11 proteins were decreased （P<0.05）.Compared with HGHL+E4 group， the survival rate of the cells in HGHL+E4+Erastin group was decreased（P<0.05）， the cell fragmentation and wrinkling were obviously decreased，the apoptotic rate and the levels of ROS and MDA were increased（P<0.05）， the levels of GSH and MMP were decreased（P<0.05）， the level of Fe²⁺ was increased（P<0.05），and the expression levels of GPX4 and SLC7A11 proteins were decreased（P<0.05）. Conclusion GLP-1R agonists can reduce the cardiomyocyte injury in the HGHL environment， and the protective effect may be related to inhibiting the iron death.

Objective To discuss the safety of secukinumab treatment during pregnancy，and to provide the reference for the treatment of psoriasis patients during pregnancy. Methods A case of pustular psoriasis safely treated with secukinumab during pregnancy was collected and combined with the relevant literature review， the safety of secukinumab treatment for the psoriasis during pregnancy was analyzed. Results The patient was a 28-year-old woman with a body weight of 50 kg. The patient had been experiencing generalized erythema and scales for 5 years， which worsened with pustules for 1 year.The dermatology examination results showed large patches of erythema， silver-white scales and small pustules on the skin of the patient， no changes in the nails， and was not accompanied by the joint symptoms. The patient was diagnosed as generalized pustular psoriasis. After being treated with secukinumab for 2 months， the psoriasis area and severity index （PASI） was 0； the patient unexpectedly became pregnant after treated for 10 months.Secukinumab was continued throughout the pregnancy until 20 d before delivery， and the patient gave birth to a healthy full-term female baby with the body weight of 3.6 kg. Conclusion Secukinumab can be used as a therapeutic option for the patients with psoriasis during pregnancy， but more clinical cases and drug research are needed to evaluate its safety during pregnancy.

Objective To discuss the expression of Klotho in placenta exosomes （Exo）of the patients with pre-eclampsia （PE），and to clarify its effect on the oxidative stress in the vascular endothelial cells. Methods The clinical data of 40 pregnant women including 20 with normal pregnancy（NP） women （NP group） and 20 PE patients （PE group） were collected. The placenta Exo in peripheral blood of the patients in two groups were isolated， and the oe-Klotho and oe-NC plasmids were transfected into the human chorionic trophoblast cells（HTR-8/SVneo）， respectively， and were regarded as oe-Klotho group and oe-NC group.The expression levels of Klotho mRNA and protein in placenta Exo and the HTR8/SVneo cells in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method and Western blotting method. The human umbilical vein endothelial cells （HUVECs） with good growth status were taken and divided into PE-Exo group （co-cultured with placenta Exo from the patients with PE），NP-Exo group （co-cultured with placenta Exo from the NP subjects）， oe-Klotho-Exo group （co-cultured with Exo from the HTR-8/SVneo cells transfected with oe-Klotho）， and oe-NC-Exo group （co-cultured with Exo from the HTR-8/SVneo cells transfected with oe-NC） according to the sources of Exo.The expression levels of exosomal marker proteins CD63， TSG101， and placenta Exo-specific marker PLAP protein were identified by transmission electron microscope （TEM） and Western blotting method； the levels of nitric oxide （NO）， reactive oxygen species （ROS）， and malondialdehyde （MDA） and the activities of superoxide dismutase （SOD） in the HUVECs in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method；the expression levels of endothelial nitric oxide synthase （eNOS） mRNA in the HUVECs in various groups was detected by RT-qPCR method；the expression levels of eNOS protein in the HUVECs in various groups were detected by Western blotting method. Results Compared with NP group， the systolic and diastolic blood pressures of the patients in PE group were increased （P<0.05）， the body weight of newborn and placenta weight were significantly decreased （P<0.05 or P<0.01）. The TEM observation results showed that the placenta Exo derived from the subjects in both NP and PE groups being round or oval vesicular discoid structures with complete membrane and similar shape， with a diameter of 50—100 nm. The Exo from the subjects in both groups highly expressed Exo marker proteins CD63 and TSG101， and placental Exo-specific protein PLAP. The nanoparticle tracking analysis （NTA） results showed that the particle size of Exo was 50—200 nm， suggesting that the vesicles isolated from the peripheral blood were derived from placental Exo. The TEM， Western blotting， and NTA results confirmed that the vesicles from oe-NC group and oe-klotho group had typical Exo structures， molecular markers， and particle size characteristics. The Exo with green fluorescence expression could be observed in the HUVECs， suggesting that the placenta Exo could be taken up by the HUVECs. The ELISA results showed that compared with NP-Exo group， the level of NO and activity of SOD in PE-Exo group were significantly decreasd（P<0.05 or P<0.01）， and the levels of ROS and MDA were significantly increased（P<0.05 or P<0.01）； compared with oe-NC-Exo group， the level of NO and activity of SOD in the HUVECs in oe-Klotho-Exo group were increased （P<0.05）， and the levels of ROS and MDA were significantly decreased （P<0.05 or P<0.01）. The RT-qPCR results showed that compared with NP group， the expression level of Klotho mRNA in placenta Exo of the patients in PE group was significantly decreased （P<0.01），and the expression level of eNOS mRNA in the HUVECs was significantly decreased （P<0.01）； compared with oe-NC group， the expression level of Klotho mRNA in Exo in the trophoblast cells in oe-Klotho group was significantly increased （P<0.01）； compared with oe-NC-Exo group， the expression level of eNOS mRNA in the HUVECs in oe-Klotho-Exo group was significantly increased （P<0.01）. The Western blotting results showed that compared with NP group， the expression level of Klotho protein in placenta Exo of the patients in PE group was significantly decreased （P<0.01）； compared with NP-Exo group， the expression level of eNOS protein in the HUVECs in PE-Exo group was significantly decreased （P<0.01）； compared with oe-NC group， the expression level of Klotho protein in Exo in the trophoblast cells in oe-Klotho group was significantly increased （P<0.01）； compared with oe-NC-Exo group， the expression level of eNOS protein in the HUVECs in oe-Klotho-Exo group was significantly increased （P<0.01）. Conclusion The placenta Exo from the PE patients may inhibit the production of NO in the HUVECs and promote the oxidative stress to impair the endothelial cell function. The over-expression of Klotho in Exo in the trophoblast cells can reduce the production of NO and level of oxidative stress in the HUVECs.

Objective To discuss the effect of the expression of stathmin 1 （STMN1） in placenta tissue of the pre-eclampsia （PE） patients on the invasion and migration functions of the trophoblast cells， and to clarify the related mechanism. Methods The placenta tissues of 17 healthy pregnant women （control group） and 15 PE patients （PE group） were selected. The human trophoblast HTR-8 cells were divided into siRNA-STMN1 group （transfected with siRNA-STMN1）， siRNA-NC group （transfected with negative control sequence siRNA-NC），pcDNA3.1-STMN1 group （transfected with pcDNA3.1-STMN1）， and pcDNA3.1-NC group （transfected with negative control sequence pcDNA3.1-NC）. The expression intensities of STMN1 and epithelial-mesenchymal transition（EMT）-related proteins in placenta tissue of the patients in two groups were observed by immunohistochemistry （IHC） staining； the proliferation rates of the HTR-8 cells in various groups were detected by CCK-8 method； the migration abilities of the HTR-8 cells in various groups were detected by cell scratch experiment； the invasion abilities of the HTR-8 cells in various groups were detected by Transwell chamber experiment； the expression levels of STMN1 mRNA in placenta tissue of the patients in two groups and HTR-8 cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） method； the expression levels of STMN1， E-cadherin， and N-cadherin proteins in placenta tissue of the patients in two groups and HTR-8 cells were detected by Western blotting method. Results Compared with control group， the systolic blood pressure， diastolic blood pressure， and the level of urine protein of the patients in PE group were significantly increased （P<0.01）， and the body weight of the newborn was increased （P<0.05）.The IHC staining results showed that compared with control group， the expression intensities of STMN1 and E-cadherin proteins in placenta tissue of the patients in PE group were increased， and the expression intensity of N-cadherin protein was decreased.The CCK-8 results showed that compared with siRNA-NC group， the proliferation rate of the HTR-8 cells in siRNA-STMN1 group was decreased（P<0.05）； compared with pcDNA3.1-NC group，the proliferation rate of the HTR-8 cells in pcDNA3.1-STMN1 group was increased（P<0.05）.The cell scratch experiment results showed that compared with siRNA-NC group，the migration rate of the HTR-8 cells in siRNA-STMN1 group was decreased（P<0.05）；compared with the pcDNA3.1-NC group， the migration rate of the HTR-8 cells in pcDNA3.1-STMN1 group was increased （P<0.05）. The Transwell chamber experiment results showed that compared with siRNA-NC group， the number of invasion cells in siRNA-STMN1 group was significantly decreased （P<0.01）； compared with pcDNA3.1-NC group， the number of invasion cells in pcDNA3.1-STMN1 group was significantly increased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression level of STMN1 mRNA in placenta tissue of the patients in PE group was decreased （P<0.05）； compared with siRNA-NC group， the expression level of STMN1 mRNA in the HTR-8 cells in siRNA-STMN1 group was significantly decreased （P<0.01）； compared with pcDNA3.1-NC group， the expression level of STMN1 mRNA in the HTR-8 cells in pcDNA3.1-STMN1 group was significantly increased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of STMN1 protein in placenta tissue of the patients in PE group was decreased （P<0.05）. Compared with siRNA-NC group， the expression levels of STMN1 and N-cadherin proteins in the HTR-8 cells in siRNA-STMN1 group were decreased （P<0.05）， and the expression level of E-cadherin protein was increased （P<0.05）. Compared with pcDNA3.1-NC group， the expression levels of STMN1 and N-cadherin proteins in the HTR-8 cells in pcDNA3.1-STMN1 group were increased （P<0.05）， and the expression level of E-cadherin protein was decreased （P<0.05）. Conclusion The expression of STMN1 in placenta tissue of the PE patients is significantly reduced，suggesting that STMN1 may be involved in the development of PE by regulating the EMT process of the trophoblast cells and inhibiting the cell proliferation， invasion， and migration.

Objective To discuss the pregnancy outcomes of the patients with different body mass index （BMI） underwent fresh embryo transfer cycles and frozen-thawed embryo transfer （FET） cycles after treated with long regimen controlled ovarian hyperstimulation （COH）， and to analyze the correlation between BMI and the pregnancy outcomes after treated with assisted reproductive technology （ART）. Methods The patients who underwent long regimen COH were divided into fresh embryo transfer cycle group （n=1 957） and FET cycle group （n=2 328）， according to their BMI， the patients in both groups were further classified into lean group（BMI <18.5 kg·m^－2）， normal group（18.5 kg·m^－2 ≤BMI <24.0 kg·m^－2）， overweight group（24.0 kg·m^－2 ≤BMI <28.0 kg·m^－2）， and obese group（BMI≥28.0 kg·m^－2）.The general informations， COH conditions， embryo conditions and clinical pregnancy outcomes of the patients in various groups were collected and analyzed. Results There were 1 957 patients， 1 957 cycles in fresh embryo transfer cycle group， which included 103 cycles in lean group， 1 254 cycles in normal group， 476 cycles in overweight group and 124 cycles in obese group. Compared with normal group， the levels of basal follicle-stimulating hormone （bFSH）， basal estradiol （bE2） and basal luteinizing hormone （bLH） of the patients in lean group were significantly increased （P<0.01）； the age of the patients in overweight group was increased （P<0.05）， the levels of bFSH， bE2， basal progesterone （bP） and bLH of the patients in overweight group were significantly decreased （P<0.01）； the levels of bFSH， bE2， and bLH of the patients in obese group were significantly decreased （P<0.01）， while the infertility time and basal testosterone （bT） level were significantly increased （P<0.05 or P<0.01）. Compared with lean group， the use time of gonadotropins（Gn） of the patients in overweight group was significantly increased （P<0.01）， the fertilization rates and transferrable embryo rates of the patients in normal， overweight and obese groups were decreased （P<0.01）. In FET cycle group， there were 1 914 patients， 2 328 cycles， including 128 cycles in lean group， 1 503 cycles in normal group， 557 cycles in overweight group and 140 cycles in obese group. Compared with lean group， the ages of patients in normal， overweight and obese groups were significantly increased （P<0.05 or P<0.01）. Conclusion For the infertile patients who are going to undergo the in vitro fertilization （IVF）， maintaining normal body weight could reduce the dosage of ovulation induction drugs， obtain more embryos， and offer more chances of pregnancy.

Objective To discuss the potential mechanism of Panax quinquefolium triolsaponins （PQTS） in the occurrence and development of ischemic stroke by using network pharmacology and molecular docking technique. Methods The potential targets of PQTS acing on IS were obtained through Swiss Target Prediction Database， Encyclopedia of Traditional Chinese Medicine（ETCM） Database， SEA Search Server Database， DisGeNET Database， and so on； the protein-protein interaction （PPI） network diagram of the key potential targets was established by STRING Database and Cytoscape 3.9.1 software；the core tagets of PQTS acting on IS were got by topology network analysis； Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were used to analyze the potential targets through DAVID online analysis website； the PQTS-target-signaling pathway network was constructed by Cytoscape 3.9.1 software and the topology network analysis was used to obtain the potential main active compositions； AutoDock Vina software was used to verify the molecular docking between the active ingredients and core targets. Results There were 122 potential targets of PQTS acting on IS； the GO function enrichment analysis was mainly included the regulation of apoptosis， intracellular signal transduction process， and regulations of extracellular substances by cells； the KEGG function analysis included the interleukins signaling pathways， phosphatidylinositol 3 kinase/protein kinase B （PI3K/Akt） signaling pathway and phosphatidylinositol 3 kinase-protein kinase B-mammalian target of rapamycin （PI3K-Akt-mTOR） signaling pathway. The molecular docking analysis results showed that pseudo-ginsenoside F11， 20（S）-protopanaxatriol， ginsenoside Rg1， ginsenoside Rh1， pseudo-ginsenoside RT5， and ginsenoside Re could form the stable conformations with signal transducer and activator of transcription 3（STAT3），phosphatidylinositol 3-kinases catalytic suburit α（PIK3CA），epidermal growth factor receptor （EGFR）， and mitogen-activated protein kinase 14（MAPK14） with lower binding energy. Conclusion The protective effect of PQTS on IS may be related to the STAT3， PIK3CA， EGFR， MAPK14， and PI3K/Akt signaling pathways.

Objective To discuss the regulatory effect of microRNA-181a-5p（miR-181a-5p） targeting BTB and CNC homology 2 （BACH2） on the apoptosis and invasion of the acute lymphoblastic leukemia （ALL） cells，and to clarify its mechanism. Methods The BACH2 over-expression recombinant plasmid（overExpBACH2）， miR-181a-5p inhibitor，and inhibitor-NC were constructed in vitro，and the transfection effect after transfected with human ALL T lymphocytes CCRF-CEM was detected by real-time fluorescence quantitative PCR （RT-qPCR） and immunofluorescence staining. The CCRF-CEM cells were divided into control group， inhibitor-NC group， miR-181a-5p inhibitor group， BACH2 over-expression（overExpBACH2） group， miR-181a-5p inhibitor+empty vector（EV） group， and miR-181a-5p inhibitor+ overExpBACH2 group. The proliferation activities of cells in various groups were detected by CCK-8 assay； the percentages of cells at G₂ phase and apoptotic rates of cells in various groups were detected by flow cytometry； the expression levels of CyclinD3， B-cell lymphoma-2（Bcl-2），Bcl-2-associated X protein（Bax）， and cysteinyl aspartate specific proteinase-3（Caspase-3） proteins in cells in various groups were detected by Western blotting method； the numbers of invasion cells in various groups were detected by Transwell chamber test； the expression levels of matrix metalloproteinase-9（MMP-9），matrix metalloproteinase 14（MMP-14）， BACH2 mRNA， and miR-181a-5p in cells in various groups were detectd by RT-qPCR method. Results Both miR-181a-5p inhibitor and overExpBACH2 vector were successfully transfected into the CCRF-CEM cells. Compared with control group，the proliferation activities of the cells in overExpBACH2 group and miR-181a-5p inhibitor+EV group were decreased （P<0.05）， and the percentages of the cells at G₂ phase and the apoptotic rates were increased （P<0.05）； the expression levels of CyclinD3 protein in the cells were decreased （P<0.05）， the expression levels of Caspase-3 protein and the ratios of Bax/Bcl-2 were increased （P<0.05）， the numbers of invasion cells were decreased （P<0.05）， the expression levels of MMP-9， MMP-14 mRNA，and miR-181a-5p were decreased （P<0.05），and the BACH2 mRNA expression levels were increased （P<0.05）. Compared with miR-181a-5p inhibitor group and overExpBACH2 group， the change trends of the above indexes in miR-181a-5p inhibitor+overExpBACH2 group were more obvious. Conclusion miR-181a-5p can target BACH2 and regulate the apoptosis and invasion of the ALL cells， which provides the new ideas for the clinical target therapy of ALL.

Objective To investigate the expression of thymosin beta 4 （Tβ4） in the breast cancer tissue and its effect on the migration and invasion of the breast cancer MCF-7 cells， and to clarify its possible mechanism. Methods Immunohistochemical staining was used to detect the expressions of Tβ4 protein in cancer tissue of 67 female breast cancer patients. The human breast cancer MCF-7 cells were divided into blank control group， siRNA-control group， and siRNA-Tβ4 group. The expression levels of Tβ4 mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method， the numbers of invasion cells in various groups were detected by Transwell chamber assay， and the scratch healing rates of cells in various groups were detected by cell scratch assay. Results Among 67 breast cancer patients， the Tβ4 was positively expressed in tumor tissue of 59 patients （88.06%）. The positive expression rates of Tβ4 was associated with the T stage， N stage， and clinical stage of the breast cancer patients （P<0.05）； there were no significant correlations between the positive expression rate of Tβ4 with patients’ age， estrogen receptor（ER） expression， human epidermal growth factor receptor-2（HER2） expression， and Ki-67 expression （P>0.05）. Compared with blank control group and siRNA-control group， the expression level of Tβ4 mRNA in the cells in siRNA-Tβ4 group was decreased（P<0.01）， the number of invasion cells was decreased（P<0.05）， and the scratch healing rate of the cells was significantly decreased（P<0.01）. Conclusion The expression level of Tβ4 in breast cancer cells is high， the expression of Tβ4 is closely correlated with the T stage， N stage and clinical stage， and targeted silencing Tβ4 could inhibit the invasion and migration abilities of the breast cancer cells.

Objective To discuss the effect of multi-kinase inhibitor motesanib combined with enhancer of zeste homolog 2（EZH2）inhibitor GSK126　on the proliferation and apoptosis of the liver cancer Huh7 cells in vitro， and to clarify its related mechanism. Methods The Huh7 cells were treated with different concentrations （0，1，5，10，20，40，and 60 μmol·L^－1）of motesanib.The proliferation rates of Huh7 cells in various groups were detected by CCK-8 assay；Western blotting method was used to detect the expression levels of EZH2 and phosphorylated EZH2（p-EZH2） proteins in the Huh7 cells in different concentrations（0，2.5，5.0，10.0，20.0 μmol·L^－1） of motesanib groups.The Huh7 cells were treated with motesanib and GSK126 in various groups；the appropriate drug concentrations were selected by CCK-8 assay and colony formation assay.The Huh7 cells were divided into control group，motesanib（10 μmol·L^－1） group， GSK126（ 5 μmol·L^－1）group， and combination （motesanib+GSK126） group. The 5-ethynyl-2'-deoxyu ridine（EdU） fluorescence staining assay was used to detect the proliferation rates of Huh7 cells in various groups；the apoptotic rates of Huh7 cells in various groups were detected by flow cytometry； Western blotting method was used to detect the expression levels of extracellular regulated protein kinases（ERK）， phosphorylated ERK （p-ERK），protein kinase B（AKT），and phosphorylated AKT（p-AKT） proteins in the Huh7 cells in various groups. Results The CCK-8 assay results showed that proliferation rate of Huh7 cells in different concentrations of motesenib groups were decreased gradually with the increasing of drug concentrations； compared with motesenib （0 μmol·L^－1） group， the proliferation rates of the Huh7 cells in 20， 40， and 60 μmol·L^－1 motesenib groups were decreased significantly （P<0.05 or P<0.01）.The Western blotting results showed that compared with 0 μmol·L^－1 motesanib group， the expression levels of EZH2 and p-EZH proteins in the Huh7 cells in 5，10， and 20 μmol·L^－1 motesanib groups were increased （P<0.05 or P<0.01）.The CCK-8 results and colony formation experiment results showed that 10 μmol·L^－1 motesanib and 5 μmol·L^－1 GSK126 could be used for the following experiment.Compared with control group，the proliferation rates of the Huh7 cells in motesanib group，GSK126 group，and combination group were significantly decreased（P<0.05 or P<0.01）； compared with motesanib group and GSK126 group， the proliferation rate of the cells in combination group was decreased （P<0.01），and the apoptotic rate was increased （P<0.01）.Compared with control group，motesanib group and GSK126 group， the expression levels of p-AKT and p-ERK proteins in the Huh7 cells in combination group were significantly decreased （P<0.05 or P<0.01）. Conclusion Motesanib alone has no significantly inhibitory effect on the proliferation and promoting effect on the apoptosis of the liver cancer Huh7 cells， which may be related to the increasing of EZH2 leading to the cell drug resistance. The combination of motesanib and GSK126 enhances the proliferation inhibition and pro-apoptotic effects of motesanib on the Huh7 cells by inhibiting AKT and ERK signaling pathways.

Objective To discuss the effect of the expression of post-meiotic segregated protein 2 （PMS2） on the biological behaviors of colon cancer SW480 cells， and to clarify the relationships between PMS2 and excision repair cross-complementation group 1（ERCC1） and extracellular regulated protein kinase （ERK） signal transduction pathway. Methods The PMS2 siRNA and PMS2 over-expression plasmids were respectively transfected into the colon cancer SW480 cells （knockdown group and PMS2 over-expression group）. The PMS2 knockdown control group （siRNA-NC group） and PMS2 over-expression control group （PMS2 control group） were wet up.Real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of PMS2 mRNA in cells in various groups； the expression levels of PMS2 protein in the cells in various groups were detected by Western blotting method； the proliferation activities of cells in various groups were detected by CCK-8 assay； cell scratch test was used to detect the migration rates of cells in various groups；the numbers of invasion cells in various groups were detected by Transwell chamber assay； the apoptotic rates of cells in various groups after cisplatin treatment were detected by flow cytometry；bioinformatics analysis was performed on the relationship between upstream and downstream proteins of PMS2， ERCC1， and ERK by using String Database；the PMS2 and ERCC1 knockdown in the SW480 cells were treated with 3 siRNA.The RT-qPCR method was used to verify the reaction between PMS2 and ERCC1；the expression levels of PMS2，extracellular regulated protein kinases 1/2（ERK1/2）， and phosphorylated ERK1/2（p-ERK1/2） proteins in the cells in various groups were detected by Western blotting method. Results The RT-qPCR and Western blotting results showed that the cell model of PMS2 gene knockdown and over-expression was successfully established. Compared with siRNA-NC group， the cell proliferative activity and cell migration rate in PMS2 knockdown group were increased（P<0.05 or P<0.01）， the number of invasion cells was increased（P<0.01），the apoptotic rate was decreased after cisplatin treatment （P<0.05）； compared with PMS2 control group， the cell proliferative activity and cell migration rate in PMS2 over-expression group were decreased（P<0.05）， the number of invasion cells was decreased（P<0.01）， and the apoptotic rate was increased after cisplatin treatment （P<0.05）.The protein-protein interaction （PPI） enrichment P value was 2.09e-07， including a total of 13 interaction nodes such as ERCC1 and ERK1/2， suggesting that there might be a regulatory effect between PMS2， ERCC1 and ERK1/2. Compared with siRNA-NC group， the expression levels of ERCC1 mRNA in the cells in various PMS2-knockout groups were significantly decreased （P<0.05 or P<0.01）. There was no significant difference in the expression of PMS2 mRNA in the cells between various ERCC1 knockdown groups and siERCC1-NC group （P>0.05）. Compared with siRNA-NC group， the expression levels of PMS2， ERK1/2， and p-ERK1/2 proteins in the cells in PMS2 knockdown group were decreased （P<0.05 or P<0.01）； compared with PMS2 control group， the expression levels of PMS2， ERK1/2，and p-ERK1/2 proteins in the cells in PMS2 over-expression group were increased （P<0.05）. Conclusion PMS2 expression can affect the proliferation， migration， invasion，and anti-apoptotic abilities of the colon cancer SW480 cells.PMS2 interacts with ERCC1 and participates in ERK signaling transduction pathway by regulating ERCC1.

Objective To observe the effect of invisible orthodontic appliance without brackets in the treatment of the patient with anterior teeth fan-shaped displacement caused by periodontitis， and to provide the clinical basis for the clinical orthodontists in the treatment of these patients. Methods One patient with anterior teeth fan-shaped displacement caused by periodontitis， received the periodontal-orthodontic combination treatment with the invisible orthodontic appliance without brackets. After the periodontal basic treatment， the patient’s periodontal condition was stabilized，The extraoral photographs， intraoral photographs，and imagological examination data of the patient before and after treatment were collected， combined with the literature review， the diagnotic plans，treatment plans， and key points of treatment of the patient were discussed. Results One 43-year-old female patient with Angle Class Ⅱ malocclusion complicated with anterior teeth fan-shaped displacement caused by periodontitis received the periodontal-orthodontic combination treatment with invisible orthodontic appliances without brackets. The extraoral and intraoral photographs before and after correction were compared， the patient’s periodontal condition was stable， and the orthodontic treatment successfully decreased the anterior teeth and closed the gap； the normal overjet and overbite and normal canine and molar relationship were established. The imagological examination before and after correction were compared， the anterior teeth of the patient were significantly adducted， and the height of the alveolar bone was basically the same as that before correction. Conclusion The periodontal- orthodontic combination treatment with invisible orthodontic appliance without brackets has a good effect in the patient with fan-shaped anterior teeth displacement caused by periodontitis， which is conducive to the long-term stability of periodontal condition of the patients.

Objective To detect the effect of quercetin （QUE） on the growth， lung metastasis，cell invasion and migration of transplanted tumor of the human lung cancer A549 cells transplanted tumor in the mice， and to clarify its related mechanism. Methods The mice were divided into control group （no intervention）， positive drug group （5 mg·kg^－1cisplatin） and low， medium and high doses （12.5， 25.0，and 50.0 mg·kg^－1 QUE）of QUE groups.Except for control group，the mice in other groups were constructed transplanted tumor models using A549 cells.The inhibitory rates of tumor weights and inhibitory rates of lung metastasis of mice in various groups were detected after treated for 10 d. The human lung cancer A549 cells were cultured in vitro and divided into normal control group， QUE group， QUE+Runt related transcription factor 3 （Runx3） negative control sequence （si-NC） （UE+si-NC） group， and QUE+Runx3 interference sequence （si-Runx3） （QUE+si-Runx3） group. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of Runt mRNA in the A549 cells in various groups； CCK-8 assay was used to detect inhibitory rates of proliferation of cells in various groups； Transwell chamber experiment was used to detect the numbers of invasion and migration cells in various groups； Western blotting method was used to detect the expression levels of Runx3， Wnt3a， β-catenin， matrix metallopeptidase-2 （MMP-2），and matrix metallopeptidase-9 （MMP-9） proteins in cells in various groups. Results Compared with control group， the inhibitory rate of tumor weight， inhibitory rate of lung metastasis and the expression levels of Runx3 protein in transplant tumor tissue of the mice in positive drug group， low， medium and high doses of QUE groups were increased（P<0.05）， and the expression levels of Wnt3a， β-catenin， MMP-2， and MMP-9 proteins were decreased （P<0.05）. Compared with normal control group，the inhibitory rate of proliferation of the A549 cells in QUE group，the expression levels of Runx3 mRNA and protein in the A549 cells in QUE group were significantly increased （P<0.05）， the numbers of invasion and migration cells were decreased （P<0.05），the expression levels of Wnt3a， β-catenin， MMP-2，and MMP-9 proteins in the cells were significantly decreased （P<0.05）. Compared with QUE+si-NC group， the inhibitory rate of proliferation of the A549 cells in QUE group，the expression levels of Runx3 mRNA and protein in the A549 cells in QUE+si-Runx3 group were significantly decreased （P<0.05）， the numbers of invasion and migration cells，the expression levels of Wnt3a， β-catenin， MMP-2，and MMP-9 proteins in the cells were significantly increased （P<0.05）. Conclusion QUE can inhibit the metastasis and invasion of transplanted tumor of the mice， and its mechanism may be related to promoting the Runx3 expression and inhibiting the Wnt/β-catenin signaling pathway， then inhibiting the invasion and migration of the lung cancer A549 cells.

Objective To analyze the effect of expression of microRNA-17-5p（miR-17-5p） in the colorectal cancer cell-derived exosomes （Exo） on the chemotherapy sensitivity of colorectal cancer cells，and to clarify its possible mechanism. Methods Real-time fluorescence quantitative PCR （qRT-PCR） method was used to detect the expression levels of miR-17-5p in the human colorectal cancer HCT116 cells， CT26 cells， LoVo cells， HT29 cells， SW620 cells， SW480 cells， and human normal colorectal mucosal epithelial HIEC cells.The CT26 cells were divided into control group，miR-NC inhibitor group， and miR-17-5p inhibitor group， and the exosomes were extracted；the morphology of the Exo was observed under transmission electron microscope； the distribution of particle size was detected by nanoparticle tracking analysis； the expressions levels of marker proteins CD9， CD63， apoptosis-inducing factor 6 interacting protein （Alix），and tumor susceptibility gene 101 （TSG101） in the Exo were detected by Western blotting method； the expression level of miR-17-5p in the Exo was detected by RT-qPCR method. The CT26 cells were divided into control group， Exo group， Exo-miR-NC inhibitor group， and Exo-miR-17-5p inhibitor group. The CT26 cells were treated with CT26 cell-derived Exo in different transfection groups， and the uptake of Exo by CT26 cells were observed by Exo green fluorescence marker PKH67 dye trace method； MTT assay was used to detect the inhibitiory rates of proliferation of the CT26 cells after treated with 1.25， 2.50， 5.00， 10.00， 20.00， and 40.00 mg·L^－1 5-fluorouracil（5-FU）； flow cytometry was used to detect the apoptotic rates of the CT26 cells in various groups； cell immunofluorescence staining was used to observe the expression intensities of microtubule-associated protein 1B light chain 3 （LC3B） in the CT26 cells in various groups； Western blotting method was used to detect the expression levels of autophagy-related proteins microtubule-associated protein light chain 3Ⅱ （LC3-Ⅱ）， microtubule-associated protein light chain 3Ⅰ （LC3-Ⅰ）， autophagy effector protein Beclin-1， and P62 proteins in CT26 cells in various groups，and the ratio of LC3-Ⅱ/LC3-Ⅰ was calculated. Results The expression levels of miR-17-5p in the human colorectal cancer HCT116， CT26， LoVo， HT29， SW620， and SW480 cells were significantly higher than that in the HIEC cells （P<0.05）； the isolated particles showed typical spherical vesicles with a peak particle size at about 140 nm， and the CD9， CD63， Alix and TSG101 proteins were all significantly expressed， indicating that Exo was successfully isolated.Compared with control group，the expression level of miR-17-5p in the Exo in miR-17-5p inhibitor group was significantly decreased （P<0.05）； compared with control group， obvious PKH67 staining could be observed around the CT26 cells in Exo group， Exo-miR-NC inhibitor group， and Exo-miR-17-5p inhibitor group， indicating that the CT26 cells could take in the Exo. Compared with control group， the inhibitory rate of proliferation and the apoptotic rate of the CT26 cells in Exo group were decreased after treated with 1.25， 2.50， 5.00， 10.00， 20.00，and 40.00 mg·L^－1 5-FU （P<0.05）， the intracellular LC3B fluorescence intensity was increased （P<0.05）， the ratio of LC3-Ⅱ/LC3-Ⅰ and the expression level of Beclin-1 protein were increased （P<0.05）， while the expression level of P62 protein was decreased（P<0.05）； compared with Exo group， the inhibitory rate of proliferation and apoptotic rate of the CT26 cells in Exo-miR-17-5p inhibitor group were increased after treated with 1.25， 2.50， 5.00， 10.00， 20.00，and 40.00 mg·L^－1 5-FU（P<0.05）， the intracellular LC3B fluorescence intensity was decreased （P<0.05）， the ratio of LC3-Ⅱ/LC3-Ⅰand the expression level of Beclin-1 protein were decreased （P<0.05），and the expression level of P62 protein was increased （P<0.05）. Conclusion The inhibition of expression of miR-17-5p in Exo derived from colorectal cancer cells can improve the chemosensitivity of the colorectal cancer cells， and the mechanism may be related to the inhibition of cell autophagy levels.

Objective To discuss the inhibitory effect of gallic acid-lecithin complex （GA-LC） on the proliferation of the lung cancer A549 cells induced by X-ray irradiation， and to clarify its possible mechanism. Methods The A549 cells were divided into different concentrations（0， 0.05， 0.10， 0.15， 0.20， 0.25， 0.30， and 0.35 g·L^－1）of GA-LC groups.CCK-8 assay was used to detect the proliferation activities of A549 cells in various groups， and the half-inhibitory concentration （IC₅₀） value of GA-LC was determined. The A549 cells were divided into control group， GA-LC group， 4 Gy X-ray irradiation group， and GA-LC+4 Gy X-ray irradiation group. CCK-8 assay was used to detect the proliferation activities of cells in various groups； flow cytometry was used to detect the reactive oxygen species （ROS） levels，mitochondrial membrane potential （MMP） levels and apoptotic rates of cells in various groups； the mitochondrial green fluorescence probe Mito-tracker was used to detect the permeability of mitochondrial membrane. Results Compared with 0 g·L^－1 GA-LC group， the proliferation activities of the cells in 0.15， 0.20， 0.25， 0.30 and 0.35 g·L^－1 GA-LC groups were significantly decreased （P<0.05）， and the IC₅₀ value of GA-LC to the A549 cells was 0.171 1 g·L^－1. Compared with control group， the proliferation activities and MMP levels of the cells in GA-LC group， 4 Gy X-ray irradiation group， and GA-LC+4 Gy X-ray irradiation group were significantly decreased （P<0.05 or P<0.01）， while the ROS levels and apoptotic rates of the cells in GA-LC group and GA-LC+4 Gy X-ray irradiation group were significantly increased （P<0.05 or P<0.01）. Compared with GA-LC group， the MMP level of the cells in GA-LC+4 Gy X-ray irradiation group was decreased（P<0.01）， while the ROS level and apoptotic rate of the cells in GA-LC+4 Gy X-ray irradiation group were significantly increased （P<0.05）； compared with 4 Gy X-ray irradiation group， the proliferation activity and MMP level of the cells in GA-LC+4 Gy X-ray irradiation group were decreased （P<0.05）， while the ROS level and apoptotic rate of the cells in GA-LC+4 Gy X-ray irradiation group were significantly increased （P<0.05 or P<0.01）. The mitochondrial fluorescence probe Mito-tracker staining results showed that the green fluorescence could be observed under fluorescence microscope； compared with control group， the green fluorescence intensities in the cells in GA-LC group， 4 Gy X-ray irradiation group， and GA-LC+4 Gy X irradiation group were increased，and the green fluorescence intensity in the cells in GA-LC+4 Gy X-ray irradiation group was the most obvious，demonstrating that the mitochondrial membrane permeability was increased. Conclusion GA-LC can enhance the inhibitory effect of X-ray irradiation on the proliferation activity of the lung cancer A549 cells， and its mechanism is related to the induction of oxidative stress，thereby increasing the mitochondrial permeability， decreasing the MMP and inducing the apoptosis.

Objective To discuss the biomechanical effect of the invisible orthodontic appliance in the remote displacement of mandibular molars assisted by the micro-implants in different parts by finite element analysis method， and to identify the optimal scheme of the micro-implant implantation site. Methods The cone beam computed tomography（CBCT） data of an Angle Class Ⅲ adult male patient with malocclusion defermity was obtained， and the Mimics Medical and 3-Matic modeling software were used to establish a three-dimensional finite element model of the remote mandibular molars with the invisible orthodontic appliance. According to whether the microimplants were used， the patients were divided into control group （without microimplants， condition 1）， and three experimental groups ［interroot micro-implant group of the first and second mandibular premolars （condition 2）， interroot micro-implant group of the second and first mandibular premolars （condition 3）， and interroot micro-implant group of the first and second mandibular molars （ condition 4）］. In the Ansys Workbench finite element analysis software， the second molar of mandible of models in various groups was moved at a step of 0.2 mm， and the molar displacement assisted by traction from the micro-implant to the invisible orthodontic appliance was applied with 2 N/side. The tooth displacement trends， deformation characteristics of the invisible orthodontic appliance，and Von Mises equivalent stress nephograms were analyzed. Results The distal and intrusive movement of the teeth to be treated were in the order of condition 4 > condition 3> condition 2> condition 1， and the distal movement of the second mandibular molar in condition 4 was 0.188 mm. In condition 1， the orthodontic teeth showed the displacement trend of mesial and labial movement， while in experimental groups， the orthodontic teeth showed the displacement trend of distal and lingual movement in the order of condition 4> condition 3 > condition 2. The extrusion deformation variable between the first molar and the second molar was the largest， and the stress peak value was 192.15 Mpa. After the stress was released， the stress concentration in control group was still located between the first molar and the second molar of the appliance， while the stress concentration in experimental groups was located on buccal surface of the appliance， and the stress peak value in condition 4 was 56.48 Mpa. Conclusion The use of micro-implant anchorage to assist the distal displacement of mandibular molars can increase the molar displacement and reduce the loss of anterior anchorage. The further back the implant site is， the more obvious the effect of molar displacement is， and the higher the tooth movement efficiency of the invivible orthodontic appliance is.

Objective To discuss the effect of baicalin on the proliferation of the human tongue squamous cell carcinoma （tongue squamous cell carcinoma） CAL27 cells， and to clarify its potential mechanism. Methods The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations （12.5， 25.0， 50.0， 100.0， and 200.0 μmol·L^－1） of baicalein groups， the clone formation of cells in various groups was observed by crystal violet staining； the proliferation rates of cells in various groups were detected by CCK-8 method；the levels of reactive oxygen species （ROS） in cells in vaious groups were detected by 2'， 7'-dichlorofluorescein diacetate （DCFH-DA） fluorescence probe； the mitochondrial membrane potential （MMP） of cells in various groups were detected by Rhodamine 123 fluorescence probe. The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations （50， 100， and 200 μmol·L^－1） of baicalin groups. Flow cytometry was used to detect the percentages of the cells at different cell cycles and the apoptotic rates of cells in various groups. The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations （50 and 100 μmol·L^－1） of baicalein groups， N-neneneba acetylcysteine （NAC） group， 50 μmol·L^－1 baicalin+NAC group，and 100 μmol·L^－1 baicalin+NAC group. The levels of ROS and MMP of cells in vraious groups were detected by DCFH-DA fluorescence probe and Rhodamine 123 fluorescence probe. Results The crystal violet staining results showed that compared with control group， the numbers of clone formation of the cells in different concentrations of baicalin groups were decreased in a concentration-dependent manner， and there was almost no clone formation of the cells in 200 μmol·L^－1 baicalin group.The CCK-8 assay results showed that compared with control group， the proliferation rates of the cells in different concentrations of baicalin groups were significantly decreased in a concentration-dependent manner（P<0.05 or P<0.01）. Compared with control group， the ROS levels of the cells in different concentrations of baicalin groups were significantly increased（P<0.05） and the MMP levels were significantly decreased （P<0.05）. Compared with control group， the percentages of the cells at S phase in different concentrations of baicalin groups were significantly increased（P<0.05）， the percentages of the cells at G₀/G₁ phase were significantly decreased （P<0.05）， and the apoptotic rates were significantly increased （P<0.05）. After the combination of baicalein and NAC， compared with control group，the ROS levels of the cells in 50 and 100 μmol·L^－1 baicalein groups were significantly increased （P<0.05） and the MMP levels were significantly decreased （P<0.05）；compared with 50 and 100 μmol·L^－1 baicalein groups， the ROS levels of the cells in 50 μmol·L^－1 baicalein+NAC group and 100 μmol·L^－1 baicalein+NAC group were significantly decreased （P<0.05），and the MMP levels were significantly increased （P<0.05）. Conclusion Baicalein can inhibit the proliferation of the CAL27 cells by activating the mitochondrial oxidative stress pathway.

Objective To discuss the improvement effect of baicalin on the myocardial hypertrophy and apoptosis induced by abdominal aorta ligation of the rats， and to clarify the potential mechanism. Methods The SD rats were randomly divided into control group， model group， low dose of baicalin （50 mg·kg^－1） group， and high dose of baicalin （100 mg·kg^－1） group.The rat model of myocardial hypertrophy was established by abdominal aorta ligation method. The rats in low and high doses of baicalin groups were given 50 and 100 mg·kg^－1 baicalin.After six weeks， the left ventricular ejection fraction （LVEF）， left ventricular posterior wall thickness at end-diastolic （LVPWd）， left ventricular posterior wall thickness at end-systolic （LVPWs），interventricular septal thickness at end-diastolic （IVSd） and interventricular septal thickness at end-systolic （IVSs） of rats in various groups were measured by echocardiography； the heart weight index（HWI） and left ventricular weight index （LVWI） of rats in various groups were calculated； HE staining was used to observe the pathomorphology of myocardium tissue of the rats in various groups； the serum levels of interleukin-8 （IL-8），interleukin-1β （IL-1β），tumor necrosis factor-α （TNF-α）， and interleukin-6 （IL-6） of the rats in various groups were detected by enzyme-linked immunosorbnent assay（ELISA） method；the expression levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax），apoptosis-inducing factor （AIF），and calpain-1 proteins in myocardium tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group， the LVEF of the rats in model group was significantly decreased（P<0.01），and the LVPWd， LVPWs， IVSd， IVSs， HWI， and LVWI were significantly increased（P<0.01）， the myocardium tissue were disordered arrangement； the serum levels of TNF-α，IL-6，IL-8， and IL-1β were increased（P<0.01）； the expression levels of Bcl-2 protein in myocardium tissue of the rats were decreased（P<0.01）， while the expression levels of Bax， AIF， and calpain-1 proteins were increased（P<0.01），and the ratio of Bax/Bcl-2 was increased（P<0.01）. Compared with model group， the LVEF of the rats in low and high doses of baicalin groups was increased（P<0.01），and the LVPWd， LVPWs， IVSd， IVSs， HWI， and LVWI were decreased（P<0.05 or P<0.01）；the cardiomyocytes in myocardium tissue were arranged in order；the serum levels of TNF-α，IL-6，IL-8， and IL-1β were decreased（P<0.05）；the expression levels of Bcl-2 protein in myocardium tissue of the rats were increased（P<0.01）， while the expression levels of Bax， AIF， and calpain-1 proteins were decreased（P<0.05）， and the ratios of Bax/Bcl-2 were decreased（P<0.05 or P<0.01）. Compared with low dose of baicalin group， the LVPWS and HWI of the rats in high dose of baicalin group were decreased（P<0.05）；the levels of TNF-α，IL-6，IL-8，and IL-1β in serum and the expression level of Bax protein were decreased（P>0.05），but there were no statistically significant differences in the other indexes（P>0.05）. Conclusion Baicalin can improve the myocardial hypertrophy and inhibit the inflammation and apoptosis induced by abdominal aorta ligation of the rats， and its mechanism may be related to the regulation of calpain-1/AIF signaling pathway.

Objective To discuss the effect of Cadherin-17 on the proliferation and apoptosis of the colorectal cancer （CRC） cells，and to clarify its possible mechanism. Methods The Cadherin-17 gene over-expression and small interference plasmids were constructed and packaged as the lentivirus and transfected into the SW480 cells to construct the stable transfection strain of over-expression and interference virus. The expression levels of Cadherin-17 mRNA and protein in the cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods， and the transfection efficiency was verified and the stable transfection strain was identified. The SW480 cells were divided into control group， empty vector group， Cadherin-17 over-expression plasmid （OV-Cadherin-17） group and Cadherin-17 small interference plasmid （si-Cadherin-17） group. The activities of cells in various groups were detected by CCK-8 assay；the apoptotic rates of cells in various groups were detected by flow cytometry； the expression levels of Cadherin-17，B-cell lymphoma-2 （Bcl-2）， Bcl2-associated X （Bax）， cytochrome c （Cyt-c） and cysteinyl aspartate specific proteinase-3 （Caspase-3），and the phosphatidylinosital-3-kinase/protein kinase B/mamalian target of repamycin （PI3K/AKT/mTOR） signaling pathway-related proteins in the cells in various groups were detected by Western blotting methods. The cells were treated with PI3K inhibitor LY294002 and divided into control group， LY294002 group， OV-Cadherin-17+LY294002 group，and si-Cadherin-17+LY294002 group； the proliferation activities and apoptotic rates of cells in various groups and the expression levels of Bcl-2，Bax，Cyt-c，Caspase-3 and the expression levels of PI3K/AKT/mTOR signaling pathway-related proteins in the cells in various groups were detected. Results The RT-qPCR and Western blotting results showed that the OV-Cadherin-17 and si-Cadherin-17 transfection and stable transfection stain were successfully constructed.Compared with control group， the proliferation ability of the cells in OV-Cadherin-17 group was increased （P<0.01）， the apoptotic rate was decreased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were decreased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were increased （P<0.01）， and the expression levels of phosphorylated PI3K（p-PI3K），phosphorylated AKT（p-Akt）， and phosphorylated mTOR（p-mTOR） proteins were increased （P<0.01）； the proliferation abilities of the cells in si-Cadherin-17 and LY294002 groups were decreased （P<0.01）， the apoptotic rates were increased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were increased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were decreased （P<0.01），and the expression levels of p-PI3K， p-AKT， and p-mTOR proteins in the cells were decreased （P<0.01）； compared with LY294002 group， the proliferation ability of the cells in OV-Cadherin-17+LY294002 group was increased （P<0.01）， the apoptotic rate was decreased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were decreased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were increased （P<0.01）， the expression levels of p-PI3K， p-AKT， and p-mTOR proteins were increased （P<0.01）， the proliferation activity of the cells in si-Cadherin-17+LY294002 group was decreased （P<0.01）， the apoptotic rate was increased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were increased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were decreased （P<0.01），and the expression levels of p-PI3K， p-AKT， and p-mTOR proteins were decreased （P<0.01）. Conclusion Cadherin-17 can promote the proliferation and inhibit the apoptosis of the CRC cells， and its mechanism may be related to the activition of PI3K/AKT/mTOR signaling pathway regulated by Cadherin-17.

Methods Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-320a in serum of the patients with actue myocardial infarction （AMI） and the myocardial H9C2 cells induced by H/R. The miR-320a inhibitor， inhibitor NC，small interference Janus kinase 2（si-JAK2）， and si-NC plasmids were transfected into the H9C2 cells respectively， and blank control group was set up. After successful transfection，the H/R treatment was performed. The H9C2 cells were divided into control group， H/R group， H/R + inhibitor NC group， H/R + miR-320a inhibitor group， H/R + miR-320a inhibitor + si-NC group and H/R + miR-320a inhibitor + si-JAK2 group. The targeting relationship between miR-320a and Janus kinase 2（JAK2） was detected by double luciferase reporter gene； the proliferation rate of cells in various groups were detected by CCK-8 assay；the activities of superoxide dismutase （SOD） and levels of malonaldehyde （MDA） in cells and the levels of lactate dehydrogenase （LDH） in cell culture supernanant in various groups were detected by biochemical method； the apoptotic rates of cells in various groups were detected by flow cytometry；the expression levels of miR-320a and JAK2 mRNA in cells in various groups were detected by RT-qPCR method；the expression levels of B-cell lymphoma-2 （Bcl-2），Bcl-2 associated X protein （Bax），cleaved-cysteinyl aspartate specific proteinase-3 （cleaved-caspase-3），JAK2， signal transducers and activator of transcription 3 （STAT3），and phosphorylated STAT3 （p-STAT3） proteins in cells in various groups were detected by Western blotting method. Results The expression levels of miR-320a in serum of the patients with AMI and the myocardial H9C2 cells in H/R group were significantly higher than those in control group （P<0.05）.The results of double Luciferase reporter gene detection suggested that miR-320a could targetedly bind with JAK2. Compared with control group， the proliferation rate of the cells and SOD activity in the cells in H/R group were decreased significantly （P<0.05），the apoptotic rate of the cells， MDA level and LDH activity in the cells were significantly increased（P<0.05），the expression levels of Bcl-2 and JAK2 proteins and ratio of p-STAT3/STAT3 in the cells were significantly decreased （P<0.05）， and the expression levels of Bax and cleaved caspase-3 proteins in the cells were significantly increased（P<0.05）.Compared with H/R group， the proliferation rate of the cells and SOD activity in the cells in H/R+miR-320a inhibitor group were increased（P<0.05），while the apoptotic rate of the cells， MDA level in the cells， LDH activity in the cell culture supernanant were decreased（P<0.05），the expression levels of Bcl-2 and JAK2 proteins and ratio of p-STAT3/STAT3 in the cells were significantly increased （P<0.05），the expression levels of Bax and cleaved- caspase-3 proteins in the cells were significantly decreased （P<0.05）. Compared with H/R+miR-320a inhibitor+si-NC group， the proliferation rate of the cells and SOD activity in the cells in H/R+miR-320a inhibitor+si-JAK2 group were decreased（P<0.05），and the apoptotic rate of the cells， MDA level in the cells，and LDH activity in the cell culture supernanant were increased（P<0.05）. Conclusion Down-regulation of miR-320a expression can inhibit the apoptosis of the cardiomyocytes induced by H/R and increase the proliferation activity of cells， and its mechanism is related to the targeted regulation of JAK2/STAT3 signaling pathway. Objective To discuss the effect of down-regulation of miR-320a expression on the myocardial hypoxia/reoxygenation （H/R） injury model， and to clarify its related mechanism.

Objective： To discuss the effect of human ceramide 1-phosphate transfer protein （CPTP） on the biological behavior of oral squamous cell carcinoma HSC-3 cells，and to clarify its related mechanism. Methods The HSC-3 cells were cultured in vitro and divided into control group and experiment group，and the cells in two groups were transfected with pFlag-CMV4 and pFlag-CPTP plasmids，respectively；the cells stably transfected with CPTP were constructed by using resistance screening method. Western blotting and immunofluorescence methods were used to detect the expression levels of CPTP protein in cells in two groups； clone formation experiment and CCK-8 assay were used to detect the numbers of clone formation and proliferation activities of cells in two groups； cell scratch experiment was used to detect the scratch healing rates of cells in two groups； Transwell chamber experiment was used to detect the numbers of invasion cells in two groups. The mice were randomly divided into control group and experiment group， and the mice were injected with the HSC-3 cells stably transfected with pFlag-CMV4 and the HSC-3 cells stably transfected with pFlag-CPTP respectively to construct the subcutaneous transplanted tumor models in the mice.The volumes and weights of transplant tumors of the mice in two groups were detected. The enrichment analysis on CPTP differentially expressed genes in head and neck squamous cell carcinoma （HNSCC） cells was analyzed by using bioinformatics method；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of p53， thrombospondin-1 （THBS1）， and cyclin G2 （CCNG2） mRNA in cells in two groups. Results Compared with control group， the expression amount of CPTP protein in the cells in experiment group was increased.Compared with control group， the number of clone formation in the cells in experiment group was significantly increased （P<0.01）， the proliferation activity and scratch healing rate of the cells were significantly increased （P<0.05 or P<0.01）， and the number of invasion cells was increased （P<0.01）. After injecting tumor cells for 2，3，and 4 weeks， compared with control group， the volumes of the transplanted tumor of the mice in experiment group were increased （P<0.05 or P<0.01）；after injecting tumor cells for 4 weeks，compared with control group，the weight of transplanted tumor of the mice in experiment group was increased（P<0.05）. The bioinformatics analysis results showed that the role of CPTP in the HNSCC tissue may be mediated through the signaling pathways such as p53. Compared with control group， the expression levels of p53， THBS1， and CCNG2 mRNA in the cells in experiment group were significantly decreased （P<0.01）. Conclusion Over-expression of CPTP can promote the proliferation， migration， invasion， and tumorigenesis of the HSC-3 cells. CPTP promotes the growth of the oral squamous cell carcinoma HSC-3 cells by inhibiting the p53 signaling pathway.

Objective： To analyze the clinical characteristics and diagnosis and treatment of the patient with primary ovarian dedifferentiated liposarcoma （DDLPS）， and to improve the clinicians’ understandings of the disease and the levels of diagnosis and treatment. Methods The clinical data of one patient with primary ovarian DDLPS was colected，and the clinicopathological manifestations， diagnosis， differential diagnosis， treatment， and prognosis were retrospectively analyzed， and the related literatures were reviewed. Results A 63-year-old female patient was admitted to hospital because of a huge mass in the lower abdomen for half a month. The gynecological ultrasound results showed there was a solid hypoechoic mass，with the size of 17.0 cm × 9.3 cm， with an irregular shape and a clear boundary in the middle pelvic cavity after hysterectomy， and there were blood flow signals in the periphery； the bilateral ovaries were not found. The whole abdominal CT results showed there was a huge mixed-density mass in the pelvic cavity， with the lobed lobes and poorly defined boundaries； the size was about 132 mm×86 mm， and the CT value was about 33 HU. The enhancement scaning results showed obvious uneven enhancement of the lesion， obvious enhancement of the edge， and no obvious enhancement of the low-density shadow in the pelvic cavity； the multiple lymph node shadows with the diameter smaller than 6 mm were seen in the pelvic cavity， and the CT enhancement scaning results showed that the lymph nodes were slightly enhancement.The tumor markers had no significant abnormalities. The patient was diagnosis as pelvic mass and the probability of ovarian malignancy was high. After completing all the relevant examinations， the transabdominal bilateral salpingectomy and oophorectomy were performed after general anesthesia. The results of the pathological diagnosis after operation were ovarian DDLPS. No recurrence of the patient was found 10 months after operation. Conclusion The primary ovarian DDLPS is rare， and the clinical manifestations are not specific； imagological methods are helpful for the diagnosis； radical surgery is the main treatment method； targeted therapy can bring good efficacy for the patients； the disease has high malignancy， poor prognosis， and it is easy to relapse； so long-term follow-up should be performed.

Objective：To establish the rat model of depressive state complicated with insomnia caused by neuropathic pain， and to clarify the effect of chronic constrict injury（CCI） on the mood and sleep-wake behaviors of the rats. Methods Sixteen male SD rats were divided into sham operation group （n=8） and CCI group （n=8）， respectively. In CCI group， 4-0 silk thread was used to ligate the right sciatic nerve trunk of the rats， and the intensity was to maintain the blood supply of the sciatic nerve adventitia. In sham operation group， the sciatic nerve trunk of the rats was exposed and separated without ligation. The paw withdraw mechanical threshold （PWMT） of rats in two groups was measured weekly after operation to evaluate the pain threshold changes. Sucrose preference test was used to detect the percentages of sucrose preference of rats in two groups； tail suspension test was used to detect the immobility time in tail suspension of rats in two groups；open field test was used to detect the total distance of horizontal movement of rats in two groups；whether or not the depression-like behaviors of the rats existed were assessed；the implantable electro encepha logram/myoelectricity was used to analyze the sleep-wake behaviors of the rats. Results Compared with sham operation group， during 1－4 weeks after modeling， the PWMT and the percentage of sucrose preference of the rats in CCI group were significantly decreased（P<0.01）， the immobility time in tail suspension of the rats was increased （P<0.01）， and the total distance of horizontal movement of the rats was decreased （P<0.01）. Compared with sham operation group，during 2－4 weeks after modeling，the subjective night wake amount of the rats in model group was increased significantly （P<0.01），the non-rapid eye movement （NREM） sleep amount was decreased （P<0.01）， and the rapid eye movement （REM） sleep amount was increased （P<0.05 or P<0.01）.Compared with sham operation group，the subjective night cumulative amount of sleep-wake states and the REM sleep cumulative amount of the rats in CCI group were increased （P<0.05 or P<0.01），and the NREM sleep cumulative amount was decreased（P<0.01）. Conclusion The CCI rats have the depressive state complicated with insomnia，and these symptoms show a chronic trend and persist until the 4th week after modeling； this model can be used for observation of long-term effect and studying of related mechanism of the depressive state comlicated with insomia.

Objective To detect the expression levels of serum and glucocorticoid-induced protein kinase 3 （SGK3） in one-cell stage fertilized eggs at different cell cycles of the mice and its subcellular localization， and to preliminarily clarify the regulatory effect of SGK3 on the early development of fertilized eggs of the mammal. Methods The one-cell stage fertilized eggs of the mice were collected by superovulation technique and in vitro culture of fertilized eggs， the expression levels of SGK3 mRNA and protein in the one-cell stage fertilized eggs at different cell cycles were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods， respectively； the subcellular localization of SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice was observed by immunofluorescence assay. Results The SGK3 mRNA and protein were expressed in the one-cell stage fertilized eggs at different cell cycles of the mice；compared with G₁， S and M phases， the expression levels of SGK3 mRNA and protein in one-cell phase fertilized eggs at G₂ phase were increased（P<0.01）. The immunofluorescence results showed that SGK3 （red fluorescence） was localized in cytoplasm of the one-cell phase fertilized eggs at G₁ and S phases， and some SGK3 entered into the nucleus at G₂ phase，and the SGK3 was widely distributed throughout the cells at M phase. Conclusion The SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice dynamically expresses， and SGK3 is shuttered between the nucleus and cytoplasm during the process of fertilized egg cell division， and the altered localization of SGK3 may drive the G₂/M transition and mitotic progression in one-cell stage fertilized eggs of the mice.

Objective： To discuss the differential expressions of calmodulin- dependent protein kinase Ⅱ （CaMKⅡ） in ovarian tissue of the normal mice and the mice with polycystic ovary syndrome （PCOS），and to clarify the effect of CaMKⅡ on the apoptosis of ovarian granulosa cells of the PCOS mice. Methods The dissolved dehydroepiandrosterone （DHEA） was used to induce the PCOS mouse models by subcutaneous injection， and the mice in control group were injected with the equal volume of castor oil. Enzyme-linked immunosorbent assay（ELISA） method was used to detect the serum testosterone level of mice in two groups； vaginal smear was used to monitor the changes of estrous cycle of the mice in two groups； HE staining was used to observe the pathomorphology of ovarian tissue of the mice in two groups； immunofluorescence staining was used to detect the localization and fluorescence intensity of CaMKⅡ protein in ovarian tissue of the mice in two groups； Western blotting method was used to detect the expression levels of CaMKⅡ and cysteine-containing aspartate protein hydrolase 3 （Caspase-3） proteins in ovarian tissue of the mice in two groups. The short hairpin RNA （sh-RNA）-CaMKⅡ lentivirus was used to transfect the human ovarian granulosa cells （KGN cells） to establish the stable transfection system. The experiment was divided into blank control， sh-CaMKⅡ-1，and sh-CaMKⅡ-2 groups.The expression levels of CaMKⅡ and Caspase-3 proteins in the KGN cells in various groups were detected by Western blotting method. Results Compared with control group， the serum total testosterone and free testosterone levels of the mice in PCOS group were significantly increased （P<0.01）， the estrous cycle was disordered and stagnated in the middle of estrus， and the ovarian tissue showed the vacuolated follicles with fewer layers of granulosa cells， indicating that the PCOS mouse model was successfully established.The CaMKⅡ protein was expressed in the oocytes， granulosa cells， and mesenchymal cells in ovarian tissue； compared with control group， the fluorescence intensity of CaMKⅡ and expression level of protein in ovarian tissue of the mice in PCOS group were significantly decreased （P<0.01）， and the expression level of Caspase-3 protein in ovarian tissue of the mice was significantly increased （P<0.01）.Compared with blank control group， the expression levels of CaMKⅡ protein in the KGN cells in sh-CaMKⅡ-1 and sh-CaMKⅡ-2 groups were significantly decreased （P<0.01）， and the expression levels of Caspase-3 protein were significantly increased （P<0.01）. Conclusion Decreasing the CaMKⅡ protein expression level in ovarian tissue of the PCOS mice induces the increasing of Caspase-3 protein expression level， thereby promoting the apoptosis of granulosa cells.

Objective To discuss the effects of different concentrations of apigenin （API） on the inflammatory and polarization response of the mouse mononuclear macrophage cells RAW264.7 induced by oxidized low-density lipoprotein （ox-LDL），and to clarify their possible mechanisms. Methods The RAW264.7 cells were divided into RAW264.7 group （without any treatment）and RAW264.7+API group （2， 4， 8， 16 and 32 μmol·L^－1 API）.After the cells were treated for 24 h.CCK-8 assay was used to detect the proliferation rates of the cells in various groups.The experimental concentration of API was selected.The RAW264.7 cells were divided into RAW264.7 group（normal RAW264.7 cells）， RAW264.7+ox-LDL group（induced with 0.08 g·L^－1 ox-LDL for 24 h）， RAW264.7+ox-LDL+low dose of API group（induced with 2 μmol·L^－1 API and 0.08 g·L^－1 ox-LDL for 24 h） and RAW264.7+ox-LDL+high dose of API group. Oil red O staining was used to observe the morphology of the foam cells in various groups. The levels of tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β）， interleukin-4（IL-4）， and interleukin-10（IL-10） in the cell culture supernant were detected by enzyme-linked immunosorbent assay（ELISA） method. The expression levels of neclear factor-κB（NF-κB） p65， phosphorylated NF-κB（p-NF-κB） p65，inducible nitric oxide synthase（iNOS）， signal transducer and activator of transcription 6（STAT6）， phosphorylated STAT-6（p-STAT6）， and arginase-1（Arg-1） in the cells in various groups were detected by Western blotting method. Results The results of CCK-8 assay showed that the proliferation rates of the cells were significantly decreased with the increasing of the concentrations of API （P<0.05）.The non-toxic low concentration （2 μmol·L^－1） and high concentration （8 μmol·L^－1） of API were selected to use in the follow-up experiment.The Oil red O staining results showed that few RAW264.7 cells were stained with Oil red O； a lot of RAW264.7 cells in RAW264.7+ox-LDL group were stained as dark red，and the lipids in the cells were increased，indicating the foam cell model was successfully established； a little of RAW264.7 cells in RAW264.7+ox-LDL+low dose of API group and RAW264.7+ox-LDL+high dose of API group were stained with Oil red O.The ELISA results showed that compared with RAW264.7 group， the levels of TNF-α and IL-1β in the cell culture supernatant in RAW264.7+ox-LDL group were significantly increased （P<0.05）， while the levels of IL-4 and IL-10 were significantly decreased （P<0.05）； compared with RAW264.7+ox-LDL group， the levels of TNF-α and IL-1β in the cell culture supernatant in RAW264.7+ox-LDL+low dose of API group，the levels of IL-4 and IL-10 were significantly increased （P<0.05）.The Western blotting results showed that compared with RAW264.7 group， the expression levels of iNOS and p-NF-κB p65 proteins in the RAW264.7 cells in RAW264.7+ox-LDL group were significantly increased （P<0.05）， and the expression levels of Arg-1 and p-STAT6 proteins were significantly decreased （P<0.05）； compared with RAW264.7+ox-LDL group， the expression levels of iNOS and p-NF-κB p65 proteins in the RAW264.7 cells in RAW264.7+ox-LDL+low dose of API group and RAW264.7+ox-LDL+high dose of API group were decreased（P<0.05），and the expression levels of Arg-1 and p-STAT6 proteins were increased（P<0.05）；compared with RAW264.7+ox-LDL+low dose of API group，the expression levels of iNOS and p-NF-κB p65 proteins in the RAW264.7 cells in RAW264.7+ox-LDL+high dose of API group were decreased（P<0.05），and the expression levels of Arg-1 and p-STAT6 proteins were increased（P<0.05）. Conclusion API can inhibit the foam cell formation of RAW264.7 cells induced by ox-LDL and improve the inflammatory response by regulating the polarization from macrophages into M2， and their mechanisms may be related to NF-κB and STAT6 pathways.

Objective To discuss the effect of Sirtuins（Sirt） protein family on the male reproductive system damage cell and animal models induced by bisphenol A （BPA）， and to clarify its effect on the male reproductive system damage induced by BPA. Methods Forty Kunming mice were randomly divided into control group， low dose of BPA group （given 3 mg·kg^－1·d^－1 BPA）， medium dose of BPA group （given 30 mg·kg^－1·d^－1 BPA）， and high dose of BPA group （given 300 mg · kg^－1·d^－1 BPA），and there were 10 mice in each group. The mice in low， medium， and high doses of BPA groups were gavaged with corn oil （0.01 mL·g^－1） mixed with corresponding concentrations of BPA， while the mice in control group were given 0.01 mL·g^－1 corn oil. After 5 weeks， the body weights， testis indexes， and epididymal indexes of the mice in various groups were detected； the sperm qualities of the mice in various groups were detected by using computer assisted semen analysis （CASA） system； the morphology of testis tissue of the mice in various groups was observed by HE staining； the expression levels of Sirt1－Sirt7 proteins in testis tissue of the mice in various groups were detected by Western blotting method. The GC-2 cells were divided into 0， 20， 40， and 80 μmol·L^－1 BPA groups （treated with 0， 20， 40，and 80 μmol·L^－1 BPA）. The proliferation rates of the GC-2 cells in various groups were detected by EdU staining； the percentages of the GC-2 cells at different cell cycles and the apoptotic rates of GC-2 cells in various groups were detected by flow cytometry；the expression levels of Sirt1－Sirt7 proteins in the GC-2 cells in various groups were detected by Western blotting method. Results Compared with control group， the testis index of the mice in high dose of BPA group was decreased （P<0.05）； compared with control group， the percentage of immobile sperm of the mice in high dose of BPA group was increased （P<0.01）， the percentage of rapid progressive sperm was decreased （P<0.01）， the percentage of medium progressive sperm was decreased （P<0.05）， and the deformity rate of the sperm was increased （P<0.01）. The HE staining results showed that the number of spermatogenic cells at all levels in the seminiferous tubules of the mice in different doses of BPA groups showed a decreasing and loosely arranged trend. Compared with control group， the expression level of Sirt6 protein in testis tissue of the mice in low dose of BPA group was decreased （P<0.01）， while the expression levels of Sirt1， Sirt2， and Sirt6 proteins in testis tissue of the mice in medium dose of BPA group mice were decreased （P<0.01），the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， Sirt5， Sirt6， and Sirt7 proteins in testis tissue of the mice in high dose of BPA group were decreased （P<0.05 or P<0.01）. The EdU staining results showed that compared with 0 μ mol·L^－1 BPA group， the proliferation rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were decreased （P<0.01）. The flow cytometry results showed that after treated for 48 h， compared with 0 μmol·L^－1 BPA group， the apoptotic rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were increased （P<0.01）. The Western blotting assay results showed that after treated for 24 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 80 μmol·L^－1 BPA group decreased （P<0.05 or P<0.01）. After treated for 48 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 40 μmol·L^－1 BPA group were significantly decreased （P<0.05 or P<0.01），and the expression levels of Sirt 5 and Sirt 6 proteins were decreased （P<0.05 or P<0.01）. Conclusion The expression levels of Sirt1－Sirt7 proteins in the male reproductive injury cells and animal models induced by BPA are decreased， which exacerbates the male reproductive system damage induced by BPA.

Objective： To observe the morphology and growth pattern of the neural stem cells neural stem cells（NSCs） in hippocampus tissue， and to provide the basis for the extraction and culture of the NSCs. Methods The NSCs were isolated from hippocampus tissue of the newborn SD rats.The morphology of the NSCs in hippocampus tissue was observed； flow cytometry was used to detect the purities of the NSCs in hippocampus tissue； immunofluorescence method was used to detect the expressions of Nestin and EdU proteins in the NSCs； the proliferative activities of the NSCs were analyzed by cell counting and CCK-8 methods. Results On the 0 day of culture，the NSCs in hippocampus tissue were suspended in the culture medium with the form of single cells；on the 2nd day of culture，the NSCs in hippocampus tissue suspended in the culture medium and began to gather into the cell clumps with different sizes and irregular shapes；On the 8th day of culture，the neurospheres showed round or oval neurosphere with different sizes， and showed clear boundary without synapse.The purity of the NSCs in hippocampus tissue was 76.50%－85.40% On the 8th day of culture，the positive expression of Nestin in the cytoplasm of the NSCs was green，and the positive expression of EdU in the nucleus was red.The neurospheres were composed of the Nestin and EdU co-staining cells. the NSCs were at logarithmic growth period，and the proliferation abilities were high when cultured for 5－11 d. Conclusion The proliferation ability of the NSCs is high；the purity of the NSCs is high on the 8th day of culture； the proliferation ability of the NSCs is high when cultured for 5－11 d.

Objective To investigate the effects of hydroxyurea （HU） combined with irradiation on the cycle cycle and apoptosis of the A549 cells after silencing α-thalassemia/mental retardation syndrome X stain-associated protein （ATRX）， and to clarify the molecular mechanism. Methods The A549 cell model （shATRX-A549） stable silencing ATRX was constructed， the infection situation of the cells was observed under fluorescence microscope，the expression of ATRX protein in the ATRX cells was detected by Western blotting method to verify the cell model， and the shNC-A549 cells were chosen as negative control. The experiment were divided into control group， HU group，radiation group （given 8 Gy X-ray radiation）， and hydroxyurea+radiation group （given HU+8 Gy X-ray radiation）.The percentages of cells at different cell cycle and apoptotic rates of the cells were detected by flow cytometry， and the expressions of mRNA in the cells after silencing ATRX were detected by RNA-sequencing （RNA-seq），and the expression amonts of the cell division cyclin 25B（CDC25B）， Cyclin B1， and cyclin dependent kinase 1（CDK1） proteins in the cells in various groups were detected by Western blotting method. Results The shNC-A549 cells and shATRX-A549 cells expressed green fluorescence protein（GFP） under fluorescence microscope；the Western blotting results showed that compared with shNC-A549 cells，the expression amount of ATRX protein in the shATRX-A549 cells was decreased. The flow cytometry results showed that compared with control group， the percentage of the shNC-A549 cells at G₀/G₁ phase in HU group was increased （P<0.05）， the percentages of the shNC-A549 cells at S phase and G₂/M phase were significantly decreased （P<0.05 or P<0.01）， and the percentages of the shNC-A549 cells at G₀/G₁ phase and S phase in radiation group were significantly decreased （P<0.01）， while the percentage of the shNC-A549 cells at G₂/M phage was significantly increased （P<0.01）， the percentage of the shNC-A549 cells at G₀/G₁ phase in HU+radiation group was significantly decreased（P<0.01）， and the percentages of the cells at S phase and G₂/M phase were significantly increased （P<0.01）.Compared with control group，the percentage of the shATRX-A549 cells at G₀/G₁ phase in HU group was increased（P<0.05），and the percentage of the shATRX-A549 cells at G₂/M phase was decreased（P<0.01）；the percentage of the shATRX-A549 cells at G₂/M phase in radiation group was significantly increased（P<0.01）， but the percentages of the shNC-A549 cells at G₀/G₁ phase and S phase were significantly decreased（P<0.01）， while the percentage of the shATRX-A549 cells at G₀/G₁ phase in HU+radiation group was decreased（P<0.01），and the percentages of the shNC-A549 cells at S phase and G₂/M phase were significantly increased （P<0.05）. Compared with the shNC-A549 cells，the percentage of the shATRX-A549 cells at G₀/G₁ phase in radiation group was increased（P<0.05），the percentage of the shATRX-A549 cells at S phase in HU+radiation group was increased（P<0.05）.Compared with control group， the apoptotic rates of the shNC-A549 cells and shATRX-A549 cells in HU group， radiation group， and HU+radiation group were significantly increased（P<0.05 or P<0.01）. Compared with shNC-A549 cells， the apoptotic rates of the shATRX-A549 cells in HU group and HU+radiation group were increased （P<0.05）. The differential expressions of mRNAs after silencing ATRX involved c-Myc， Esp1， Cdc20， Plk1， CycA/B， Cip1， and PCNA.The Western blotting results showed that compared with control group， the expression amounts of CDC25B， Cyclin B1， and CDK1 proteins in the shNC-A549 cells and shATRX-A549 cells in HU group， radation group and HU+radiation group were decreased；compared with the shNC-A549 cells， the expressions amounts of Cyclin B1 protein in the shATRX-A549 cells in control and HU groups were decreased， while the expression amounts of CDC25B，cyclin B1， and CDK1 proteins in the shATRX-A549 cells in radiation group and HU+radiation group were increased. Conclusion HU and radiation can cause the cell cycle arrest and the apoptosis of the A549 cells after silencing ATRX， and its mechanism is related to the CDC25B/Cyclin B/CDK1 pathway.

Objective To analyze the quantitative constitutive relationship between the median-inhibitory concentration （IC₅₀） of halobenzoquinones （HBQs） to the human hepatocellular carcinoma HepG2 cells and structural parameters， and to explore the structural parameters that may affect the cytotoxicity of HBQs. Methods The HepG2 cells were cultured in vitro and cultured with different contentrations of HBQs solutions，at the same time， blank control group ［10% fetal bovine serum（FBS）+DMEM culture medium］ and negative control group ［（10% FBS+DMEM culture medium +1.33% methanol （MeOH）］ were set up. The action concentrations of HBQs were comfirmed with MTS and neural red uptake（NRU） methods，respectively.The survival rates of cells in various groups were detected by MTS and NRU methods， and the IC₅₀ of HBQs was calculated. The quantum chemical calculation module MOPAC was used to calculate the structural parameters of every kind of HBQs. Single-factor linear regression analysis was used to construct the quantitative structure-toxicity relationship （QSTR） models based on the IC₅₀ and structural parameters of the HBQs. Results The MTS method determination results showed that the action concentrations of 2，6-dichloro-1，4-benzoquinone（2，6-DCBQ） were 75， 100， 125， 150， 175， 200， 225， and 250 μmol·L^－1，the action concentrations of 2，6-dichloro-3-methyl-1，4-benzoquinone（DCMBQ） were 175， 200， 225， 250， 275， 300，and 325 μmol·L^－1， the action concentrations of 2，3，6-trichlorocyclohexa-1，4- benzoquinone（TCBQ） were 200， 225， 250， 275， 300， and 325 μmol·L^－1， the action concentrations of 2，5-dibromo-1，4-benzoquinone（2，5-DBBQ） were 75， 100， 125， 150， 175， and 200 μmol·L^－1，and the action concentrations of 2，6-dibromo-1，4-benzoquinone（2，6-DBBQ） were 200， 225， 250， 275， 300， 325 μmol·L^－1.The NRU method determination results showed that the action concentrations of 2，6-DCBQ were 25， 50， 75， 100， and 125 μmol·L^－1， the action concentrations of DCMBQ were 125， 150， 175， 200， 225，and 250 μmol·L^－1， the action concentrations of TCBQ were 175， 200， 225， 250， 275，and 300 μmol·L^－1，the action concentrations of 2，5-DBBQ were 75， 100， 125， 150，and 175 μmol·L^－1，the action concentrations of 2，6-DBBQ were 125， 150，175， 200， 225， and 250 μmol·L^－1.The survival rates of HepG2 cells in various groups were significantly decreased with the increase of HBQs concentration， showing a dose-response relationship. The cytotoxic effects of HBQs on the HepG2 cells detected by MTS method were 2，6-DCBQ>2，5-DBBQ>DCMBQ>TCBQ>2，6-DBBQ；and the cytotoxic effects of HBQs on HepG2 cells detected by NRU method were 2，6-DCBQ>2，5-DBBQ>DCMBQ>2，6-DBBQ>TCBQ.The single-factor multiple linear regression analysis results showed that the correlation analysis between IC₅₀ and structural parameters of the HBQs detected by MTS and NRU methods had no statistically significant differences（P>0.05）. Conclusion The substitution position， type and number of halogen atoms can affect the toxic effects of HBQs， and there is a trend that the more halogen substitution， the less toxic it is.

Objective To discuss the antioxidant capacities of ten edible plant exosomes-like nanoparticles （ELNs） from ginger， green pepper， garlic， mushroom， lemon， yam， grapes， tomatoes， broccoli，and onions in vitro and their protective effects on oxidative injury of the PC12 cells induced by hydrogen peroxide，and to provide the basis for the further study on the ELNs. Methods The PC12 cells were divided into control group， exosome group， hydrogen peroxide group，and hydrogen peroxide+exosomes group. Ten types of ELNs were separated and extracted by using high-speed differential centrifugation. The in vitro antioxidant capacities of these 10 types of ELNs were detected by using 1，1-diphenyl-2-picryl-hydrazyl（DPPH） free radical scavenging system，2，2'-azinobis-（3-ethylbenzthiazoline-6-sulphonate（ABTS） cationic radical system， and ferric ion total reduction antioxidant power（FRAD） system. The viabilities of PC12 cells in various groups were detected by MTT method. Results The DPPH radical scavenging abilities of ten ELNs，from high to low， were mushroom， onion， ginger， lemon， grape， tomato， green pepper， broccoli， yam， and garlic； the capacities of ten ELNs to scavenge ABTS， from high to low， were ginger， mushroom， onion， lemon， yam， grape， green pepper， garlic， tomato， and broccoli； the total antioxidant capacities of ten ELNs in FRAP， from high to low， were ginger， green pepper， onion， mushroom， lemon， broccoli， grape， yam， tomato，and garlic. The antioxidant capacities of five ELNs with stronger antioxidant capacities were increased with the increasing concentration of the ELNs in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The half maximal inhibitory concentration（IC₅₀） values of onion were 73.15， 123.02， and 83.00 g·L^－1， the IC₅₀ values of ginger were 124.07， 91.24， and 91.24 g·L^－1， the IC₅₀ values of mushroom were 310.44， 518.04，and 237.10 g·L^－1， the IC₅₀ of green pepper were 969.06， 847.32，and 237.10 g·L^－1 and the IC₅₀ values of lemon were 1 718.94， 544.38，and 962.12 g·L^－1； compared with control group， the viability of the PC12 cells in hydrogen peroxide group was decreased by about 30%. Compared with hydrogen peroxide group， the viability of the PC12 cells in hydrogen peroxide+ELNs groups （green pepper， lemon， yam， and broccoli） were increased by 21.08%， 26.2%， 11.72%， and 15.15%. Conclusion The ELNs from ginger， mushroom， lemon， and onion show stronger antioxidant capacities in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The ELNs from green pepper， lemon， yam， and broccoli have protective effects on the oxidative damage of the PC12 cells induced by hydrogen peroxide.

Objective To discuss the clinical manifestations and diagnosis and treatment processes of one patient with subintimal hemangioma of the knee joint， and to improve the clinicians’ understandings of the disease. Methods The clinical data， imagological data， arthroscopic manifestations， and pathological results of one patient with subsynovial hemangioma of the knee joint were retrospectively analyzed， and the related literatures were reviewed. Results A 22-year-old female patient presented with intermittent left knee joint pain and swelling for 7 years. The physical examination results showed obvious tenderness in the inner and outer spaces of the left knee joint， and the range of motion of the left knee joint was decreased （0°－90°）.The ultrasound results showed that the hypoechoic light clusters were found between the deep layer of the vastus medialis tendon and the synovial tissue； the magnetic resonance imaging（MRI） results showed that the deep medial retinaculum of the left knee joint was swollen near the parapatellar soft tissue，which showed a slightly low signal on T1-weighted sequences and a patch-like signal with slightly higher intensity on T2 fat-suppressed sequences；the high T2 signal was seen in the synovial tissue， and the boundary was not clear. The preliminary consideration was left knee mass. The arthroscopic left knee joint lesion resection was performed； under the arthroscope， the appearance of the medial parapatellar synovium was basically normal，after removing the intimal layer of the superficial synovial tissue， the mass was located between the subintima of the synovium and the joint capsule， the mass was completely removed for submission， and the pathology results showed it was subsynovial hemangioma of the knee joint； the original pain in the left knee joint of the patient disappeared after operation. Conclusion Lots of patients with subsynovial hemangioma of the knee joint have medical history of trauma， manifesting as unexplained pain， swelling， and limited mobility of the knee joint； the diagnosis of the disease needs to be confirmed through the combination of imagological manifestations， arthroscope， and pathological examination results；the arthroscope surgery can significantly improve the prognosis of the patients， and it has some advantages，such as fewer surgical injuries， fewer postoperative complications， and faster recovery.

Objective To discuss the effect of long non-coding RNA （lncRNA） lung cancer metastasis-associated transcript 1 （MALAT1） on the angiogenesis of the human brain microvascular endothelial cells （HBMECs） induced by oxygen-glucose deprivatioin/reoxygenation（OGD/R）， and to clarify its potential molecular mechanism. Methods The binding sites of MALAT1， heterogeneous nuclear ribonucleoprotein K （hnRNPK）， and vascular endothelial growth factor A （VEGFA） were predicted by bioinformatics. The HBMECs were treated with OGD/R to establish the cerebral ischemia cell model. The HBMECs were divided into control group， OGD/R model group， OGD/R+siNC group， OGD/R+ silencing MALAT1 （OGD/R+siMALAT1） group and OGD/R+siMALAT1+over-expression of VEGFA （OGD/R+siMALAT1+VEGFA） group.Small interference RNA（siRNA） was used to silence the expression of MALAT1， and pcDNA vector was used to construct the VEGFA over-expression vector. The constructed siMALAT1 and pcDNA VEGFA vectos were transfected into the HBMECs separately or simultaneously. The angiogenesis abilities of cells in various groups were detected by tube formation assay； the expression levels of VEGFA protein in the cells in various groups were detected by Western blotting method. Twenty 6-week-old healthy male C57BL/6 J mice were randomly divided into sham operation group， middle cerebral artery occlusion （MCAO） model group， MCAO+NC blank vector（MCAO+NC） group， and MCAO+ over-expression of MALAT1 （MCAO+MATAL1） group，and there were 5 mice in each group. Except for sham operation group， the mice in the other groups were used to construct the MCAO mouse models by suture method， and the NC empty vector and MALAT1 over-expression vector were injected into the right ventricle of the mice in MCAO+NC group and MCAO+MATAL1 group， respectively. The percentage of cerebral infarction area of mice in various groups were detected by 2，3， 5-triphenyltetrazole chloride （TTC） staining，and the expression levels of VEGFA protein in brain tissue of mice in various groups were detected by Western blotting method. Results The bioinformatics results showed that there were binding sites among MALAT1 and hnRNPK， hnRNPK and VEGFA. The results of tube formation experiment and Western blotting showed that compared with control group， the number of the tubes in OGD/R model group was significantly increased （P<0.01）， and the expression level of VEGFA protein in the cells in OGD/R model group was significantly increased （P<0.01）； compared with OGD/R model group， the number of the tubes in OGD/R+siMALAT1 group was significantly decreased （P<0.01）， and the expression level of VEGFA protein in the cells in OGD/R+siMALAT1 group were significantly decreased （P<0.01）； compared with OGD/R+siMALAT1 group， the number of the tubes in OGD/R+siMALAT1+VEGFA group was significantly increased （P<0.01）， and the expression level of VEGFA protein in the cells in OGD/R+siMALAT1+VEGFA group was significantly increased （P<0.01）. In the MCAO model mouse experiment， the results of TTC staining and Western blotting showed that compared with sham operation group， the percentage of cerebral infarction area and the expression level of VEGFA protein in brain tissue of the mice in MCAO model group were significantly increased （P<0.01）； compared with MCAO model group， the percentage of cerebral infarction area of the mice in MCAO+MALAT1 group was significantly decreased （P<0.01）， and the expression level of VEGFA protein in brain tissue of the mice in MCAO+MALAT1 group was significantly increased （P<0.01）. Conclusion LncRNA MALAT1 can enhance the stability of the target gene VEGFA and upregulate its expression， and promote the angiogenesis of the cerebral ischemic stroke mice， which provides a new direction for the treatment of cerebral ischemic stroke.

objective To prepare a composite photocrosslinked hydrogel containing zeolite imidazole framework-8 （ZIF-8），and to evaluate its in vitro cytotoxicity， drug release capability， and antimicrobial propertie. Methods The ZIF-8 particles were synthesized by hydrothermal method， and the microstructure characteristic was observed under scanning electron microscope （SEM）. The particles were mixed with the gelatin methacryloyl （GelMA） with the mass fraction of 0.2% to obtain the composite hydrogel GelMA-Z. The atomic absorption spectroscope was used to detect the cumulative zinc ion （Zn²⁺） release amounts in GelMA-Z at different time points.The NIH-3T3 cells were co-cultured with GelMA-Z for 1， 3， and 7 d；the viabilities of the cells in various groups were detected by CCK-8 assay； the GelMA-Z was co-cultured with Escherichia coli （E. coli） and Staphylococcus aureus （S. aureus） for 6，12，and 24 h and divided into control group， GelMA group， and GelMA-Z group.The bacterial activities of the cells in various groups at different time points were detected by microplate reader； the bacterial formation and the presence of live/dead becterial staining condition were detected by plate antibacterial experiment and live/dead bacterial staining method. Results The SEM observation results showed that the hydrothermally synthesized ZIF-8 particles had the uniform particle sizes.The atomic absorption spectroscope results showed that Zn²⁺ in GelMA-Z showed an initial burst phase within 1 d， followed by a slow release，and reached the equilibrium around 7 d.Compared with control group，the viabilities the cells in GelMA group and GelMA-Z group were above 90% on the 1st， 3rd， and 7th days， but there was no significant difference（P>0.05）.The bacterial activity detection results showed that when co-cultured with bacteria for 6，12，and 24 h， compared with control group and GelMA group，the bacterial activities of the E. coli and S. aureus in GelMA-Z group were decreased （P<0.05）. The plate antibacterial experiment results showed that the number of bacterial formation in GelMA-Z group was fewer than those in control group and GelMA group. The live/dead bacterial staining results showed that in GelMA-Z group，there was a large number of red fluorescence stained dead bacteria； in control group and GelMA group， there was a large number of green fluorescence stained live bacteria. Conclusion The GelMA hydrogel loaded with ZIF-8 particles can achieve the in situ photocrosslinking and possesses good Zn²⁺ release capability and antimicrobial activity， and it is a novel hydrogel dressing for treatment of the infected wounds.

Objective To discuss the effect of honeysuckle extract on the proliferation and apoptosis of airway smooth muscle cells （ASMCs） in the asthmatic mice，and to clarify the related mechanism. Methods The asthma models of mice were constructed， and the primary ASMCs were isolated and identified. The M2 polarization of macrophages RAW264.7 was induced and identified. The RAW264.7 cells were divided into control group， low， medium and high doses of honeysuckle extract groups. The RAW264.7 cells in control group were induced M2 polarization and co-cultured with ASMCs. The RAW264.7 cells in low， medium， and high doses of honeysuckle extract groups were treated with different concentrations （50， 100， and 200 mg·L^－1） of honeysuckle extract for 1 h and then treated with interleukin-4 （IL-4） （60 μg·L^－1） for 6 h. Then the RAW264.7 cells and ASMCs were co-cultured for 24 h. The percentages of CD86 and CD206 positive cells in various groups were detected by flow cytometry； the levels of interleukin-10（IL-10） in culture supernatant of the ASMCs in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method； the proliferation activities of cells in various groups were detected by MTT assay； the apoptotic rates of ASMCs in various groups were detected by flow cytometry. Results The morphology of the cells and immunofluorescence staining results proved that the extracted cells were the ASMCs.The M2 polarization identification results showed that the RAW264.7 cells were induced into the M2 macrophages.Compared with control group， the percentages of CD86 positive cells in the ASMCs in low， medium and high doses of honeysuckle extract groups were significantly increased （P<0.05），while the percentages of CD206 positive cells were decreased（P<0.05）；the IL-10 levels in culture supernatant of the ASMCs and the proliferation activities of ASMCs were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）. Compared with low dose of honeysuckle extract group， the percentages of CD86 positive cells in medium and high doses of honeysuckle extract groups were significantly increased（P<0.05）， while the percentages of CD206 positive cells were decreased（P<0.05）；the IL-10 levels in the culture supernatant and proliferation activities of the cells were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）.Compared with medium dose of honeysuckle extract group， the percentages of CD86 positive cells in high dose of honeysuckle extract group was significantly increased（P<0.05）， while the percentage of CD206 positive cells was decreased（P<0.05）； the IL-10 levels in the culture supernatant and proliferation activities of the cells were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）. Conlusion： Honeysuckle extract can inhibit the proliferation and promote the apoptosis of the ASMCs in the asthmatic mice by inhibiting the M2 polarization of the macrophages.

Objective： To discuss the improvement effect of β-sitosterol on the cognitive function of the Alzheimer’s disease （AD） model mice， and to elucidate its possible molecular mechanism. Methods Sixty mice were randomly divided into control group， sham operation group， AD group， donepezil hydrochloride （0.6 mg·kg^－1）group， low dose （1.0 mg·kg^－1） of β-sitosterol group， and high dose （4.0 mg·kg^－1）of β- sitosterol group； and there were 12 mice in each group. The mice were injected with β amyloid protein 1-42 （Aβ_1-42） to establish the AD models，and β- sitosterol interventional treatment was conducted. The free activity numbers of the mice in various groups were observed by autonomous activity experiment； the scores of nesting behavior of the mice in various groups were observed by nesting behavior experiment；the identification time of new objects and the discrimination ratios （DR） of new and old objects of the mice in various groups were observed by new object discrimination experiment； Morris water maze experiment was used to observe the escape latencyies and numbers of crossing platform of the mice in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of interleukin-17A（IL-17A） and p53 mRNA in hippocampus tissue of the mice in various groups； Western blotting method was used to detect the expression levels of IL-17A， p53，Caspase-3， cleaved Caspase-3，B-cell lymphoma 2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in hippocampus tissue of the mice in various groups；enzyme-linked immunosorbent assay （ELISA） was used to detect the activities of superoxide dismutase （SOD） and levels of glutathione （GSH），tumor necrosis factor α（TNF-α）， and IL-17A in hippocampus tissue of the mice in various groups. Results The nesting behavior experiment results showed that compared with control group， the score of nesting behavior of the mice in AD group was significantly decreased （P<0.01）； compared with AD group， the scores of nesting behavior of the mice in donepezil hydrochloride group and low dose of β-sitosterol group were increased （P<0.05）. The new object discrimination experiment results showed that compared with control group， the identification time of new objects of the mice in AD group was decreased （P<0.01），and DR was decreased（P<0.01）； compared with AD group，the identification time of new objects of the mice in donepezil hydrochloride group and high dose of β-sitosterol group were significantly increased （P<0.01）， and DR was increased （P<0.05）. The Morris water maze experiment results showed that on the 3rd and 5th days after training， compared with control group， the escape latency of the mice in AD group was significantly increased （P<0.01）； compared with AD group， the escape latencies of the mice in latendonepezil hydrochloride group and low and high doses of β-sitosterol groups were increased （P<0.05 or P<0.01）. Compared with control group， the number of crossing platforms of the mice in AD group was increased （P<0.01）； compared with AD group， the numbers of crossing platform of the mice in donepezil hydrochloride group and low and high doses of β-sitosterol groups were decreased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of IL-17A and p53 mRNA in hippocampus tissue of the mice in AD group were significantly increased （P<0.01）； compared with AD group，the expression levels of IL-17A mRNA in hippocampus tissue of the mice in low and high doses of β-sitosterol groups were decreased （P<0.05）， while the expression levels of p53 mRNA in hippocampus tissue of the mice in donepezil hydrochloride group and high dose of β-sitosterol group were significantly decreased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of IL-17A， p53， and cleaved Caspase-3 proteins and the Bax/Bcl-2 ratio in hippocampus tissue of the mice in AD group were significantly increased （P<0.01）； compared with AD group， the expression levels of IL-17A and p53 proteins and Bax/Bcl-2 ratio in hippocampus tissue of the mice in low dose of β-sitosterol group were significantly decreased （P<0.05 or P<0.01）， while the expression levels of IL-17A and p53 proteins and Bax/Bcl-2 ratio in hippocampus tissue of the mice in donepezil hydrochloride group and high dose of β- sitosterol group were decreased （P<0.01）. The ELISA results showed that compared with control group， the SOD activity and GSH level in hippocampus tissue of the mice in AD group were decreased （P<0.01）， while the levels of TNF-α， IL-17A and IL-17A in hippocampus tissue of the mice were increased （P<0.01）； compared with AD group， the SOD activity and GSH level in hippocampal tissue of the mice in high dose of β- sitosterol group were significantly increased （P<0.01），while the IL-17A level was decreased （P<0.05）， and the level of TNF-α in hippocampus tissue of the mice in low dose of β-sitosterol group was decreased （P<0.05）. Conclusion β-sitosterol can reduce the neuroinflammation and inhibit the apoptosis，thereby improving the cognitive function of the AD model mice；its mechanism may be related to inhibiting the IL-17-p53 signaling pathway.

Objective To discuss the potential categories of adolescents’ negative physical intentions， and to further analyze the relationships between different potential categories and suicidal ideation and non-suicidal self-injury of adolescents’ negative physical intentions. Methods In Changchun City of Jilin Province，4 middle schools were randomly selected， and 2 or 3 teaching classes were selected from each grade by using cluster randomly sampling method，and 1 459 students were regarded as the survey subjects， Beck Scale for Suicide Ideation Chinese Version（BSI-CV）， Adolescent Self Harm Scale （ASHS）， and Negative Physical Self Scale （NPSS） were used for the on-the-spot questionnaire survey， and the results were analyzed by latent profile analysis，χ² test，and Logistic regression analysis. Results The adolescents’ negative physical intentions were divided into body satisfaction（65.2%，951 persons），emaciated body dissatisfaction（13.4%，195 persons） and obese appearance dissatisfaction（21.4%，313 persons）.The Logistic regression analysis results showed that only-child（OR=2.43，95%CI：1.21－4.89，P<0.05）， bullying experience（OR=2.43， 95%CI：2.19－5.72，P<0.05），emaciated body dissatisfaction（OR =5.42， 95%CI：2.66－11.05，P<0.01），and obese appearance dissatisfaction（OR=9.34， 95%CI：5.18－16.83，P<0.01） were the risk factors for the adolescents’ suicidal ideation； girls（OR=2.35，95%CI：1.49－3.71，P<0.01），single-parent family（OR=1.99，95%CI：1.11－3.59，P<0.05），bullying experience（OR=5.26， 95%CI：3.42－8.08，P<0.01），emaciated body dissatisfaction（OR=2.34， 95%CI：1.21－4.53，P<0.05），and obese appearance dissatisfaction（OR=5.24， 95%CI：3.31－8.28，P<0.01）were the risk factors for the non-suicidal self-injury of the adolescents. Conclusion The negative physical intentions of the adolescents have heterogeneous；emaciated body dissatisfaction and obese appearance dissatisfaction are the risk factors for the suicidal ideation and non-suicidal self-injury.

Objective To discuss the effect of Jiegeng Yuanshen Tang（JGYST） on airway tissue inflammation and mucus secretion in the mice with allergic asthma， and to clarify the related mechanism. Methods Forty male C57BL/J mice were randomly divided into control group， JGYST group， ovalbumin （OVA） group， and OVA + JGYST group. The mice in OVA group and OVA +JGYST group were sensitized with 50 μg OVA via intraperitoneal injection twice weekly， followed by 20 μg OVA nasal drops daily for 7 d to induce asthma；the mice in OVA +JGYST group were gavaged with 200 μL JGYST 1 h before each OVA challenge， and the administration lasted for 7 d； the mice in control group were given equivalent dose of PBS via intraperitoneal injection， nasal drops， and gavage； the mice in JGYST group were given the same dose of PBS for intraperitoneal and nasal administration and gavaged with the same dose of JGYST. The pathomorphology of lung tissue of the mice in various groups was observed by HE staining and periodic acid-Schiff （PAS） staining， and the inflammation and PAS scores were calculated； flow cytometry method was used to detect the numbers of eosinophils， neutrophils， helper T lymphocyte 1 （Th1） cells， helper T lymphocyte 2 （Th2） cells， and dendritic cells （DCs）， as well as the percentage of mature DCs and level of reactive oxygen species （ROS） in lung tissue of the mice in various groups；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of interleukin-4 （IL-4）， interleukin-10 （IL-10）， and tumor necrosis factor-α （TNF-α） mRNA in lung tissue of the mice in various groups. Results The HE and PAS staining results showed that the mice in control group had intact airway and alveolar structure，without infiltration of inflammatory cells or mucus secretion； compared with control group， there was a large number of infiltrating inflammatory cells in airway tissue of the mice in OVA group，and the inflammation and PAS scores were increased （P<0.01）； compared with OVA group， the infiltration of inflammatory cells in airway tissue of the mice in JGYST group and OVA + JGYST group was decreased， and the inflammation and PAS scores were significantly decreased （P<0.01）. The flow cytometry results showed that compared with control group， the numbers of eosinophils， Th2 cells， and DCs in lung tissue of the mice in OVA group were increased （P<0.05 or P<0.01）， and the percentage of mature DCs and level of ROS were significantly increased （P<0.01）； compared with OVA group， the numbers of eosinophils， Th2 cells， and DCs in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased （P<0.01）， and the percentage of mature DCs and level of ROS were significantly decreased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of IL-4， IL-10， and TNF-α mRNA in lung tissue of the mice in OVA group were increased （P<0.01）； compared with OVA group， the expression levels of IL-4 and TNF-α mRNA in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased （P<0.01）， while the expression level of IL-10 mRNA was increased （P<0.01）. Conclusion JGYST can alleviate the airway tissue inflammation and mucus secretion in the mice with allergic asthma，and its mechanism may be related to reducing the number of Th2 cells and DCs， decreasing the ROS level and expression level of proinflammatory cytokine， and increasing the expression level of anti-inflammatory cytokine.