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 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 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 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 improvement effect of hydatid antigen B （HA-B） on the immune thrombocytopenia （ITP） of the mice， and to clarify its related mechanism. Methods The C57/B6 mice with wild-type （WT） and tumor necrosis factor receptor 2 （TNFR2） gene knockout （TNFR2^－/－） were divided into WT control group， WT-ITP model group， WT-HA-B group， TNFR2^－/－control group， TNFR2^－/－ ITP model group， and TNFR2^－/－ HA-B group. The body weights， organ indexes， and blood routine indexes of mice in various groups were detected. The percentages of M2 macrophages in peripheral blood of mice in various groups were detected by flow cytometry； enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of Arginase 1 （Arg1）， interleukin-10 （IL-10），inductible nitric oxide synthase （iNOS），and interleukin-6 （IL-6） in serum of the mice in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of Arg1， IL-10， iNOS and IL-6 in bone marrow derived macrophages （BMDM） of the mice in various groups. The BMDM of the mice in WT control group and TNFR2^－/－ control group were induced into the M2 macrophages （WT M2 group and TNFR2^－/－ M2 group）. After adding HA-B （WT M2+HA-B group and TNFR2^－/－ M2+HA-B group）， the expression levels of Arg1 and IL-10 mRNA in the cells in various groups were detected by RT-qPCR method. Results Compared with WT control group and TNFR2^－/－ control group， the body weights of the rats in WT-ITP model group and TNFR2^－/－ ITP model group were decreased （P<0.05）， the spleen and thymus indexes were increased （P<0.05）， the platelet counts and red blood cell counts were decreased （P<0.05）， the hemoglobin levels were decreased （P<0.05）， the white blood cell counts were increased （P<0.05）， and the coagulation time were increased（P<0.05）； the percentages of M2 macrophages in the peripheral blood were decreased （P<0.05）， the levels of Arg1 and IL-10 in serum were decreased （P<0.05）， and the levels of iNOS and IL-6 were increased （P<0.05）； the expression levels of Arg1 and IL-10 mRNA in the BMDM were decreased （P<0.05）， while the expression levels of iNOS and IL-6 mRNA were increased （P<0.05）. Compared with WT-ITP model group， the body weight of the mice in WT-HA-B group was increased （P<0.05）， the spleen and thymus indexes were decrease （P<0.05）， the platelet count and red blood cell count were increased （P<0.05）， the hemoglobin level was increased （P<0.05）， the white blood cell count was decrease （P<0.05）， and the coagulation time was decreased （P<0.05）； the percentage of M2 macrophages in the peripheral blood was increased （P<0.05）， the levels of Arg1 and IL-10 in the serum were increased （P<0.05）， and the levels of iNOS and IL-6 in serum were decreased （P<0.05）；the expression levels of Arg1 and IL-10 mRNA in the BMDM were increased（P<0.05）， while the expression levels of iNOS and IL-6 mRNA were decreased （P<0.05）. Compared with WT-HA-B group， the body weight of the mice in TNFR2^－/－ HA-B group was decreased （P<0.05）， the spleen and thymus indexes were increased （P<0.05）， the platelet count and red blood cell count were decreased （P<0.05）， the hemoglobin level was decreased （P<0.05）， the white blood cell count was increased （P<0.05）， and the coagulation time was increased （P<0.05）； the percentage of M2 macrophages in the peripheral blood was decreased （P<0.05）， the levels of Arg1 and IL-10 in the serum were decreased （P<0.05）， and the levels of iNOS and IL-6 were increased （P<0.05）； the expression levels of Arg1 and IL-10 mRNA in the BMDM were decreased （P<0.05）， while the expression levels of iNOS and IL-6 mRNA were increased （P<0.05）. Compared with WT M2 group， the expression levels of Arg1 and IL-10 mRNA in the macrophages in WT M2+HA-B group were increased （P<0.05）； compared with WT M2+HA-B group， the expression levels of Arg1 and IL-10 mRNA in the M2 macrophages in TNFR2^－/－M2+HA-B group were decreased （P<0.05）. Conclusion HA-B can promote the macrophage M2 polarization through TNFR2， thereby exerting the therapeutic effect on ITP.

Objective To discuss the effect of 20 （S） - protopanaxadiol （PPD） on the osteogensis differentiation of bone marrow mesenchymal stem cells （BMSCs） of the rats， and to clarify the osteogenesis induction mechanism. Methods The BMSCs were extracted from the SD rats by whole bone marrow method，and the BMSCs were divided into control group and 2.5， 5.0， 10.0， 20.0， and 40.0 mg·L^－1 PPD groups； CCK-8 assay was used to detect the proliferation activities of BMSCs in various groups；the survival of BMSCs in various groups was observed by Calcein/PI staining；the suitable concentration of PPD for the subsequent experiments was selected. The BMSCs were divided into control group and PPD group. On the 7th day of osteogenesis induction， BCIP/NBT colorimetry was used to carry out alkaline phosphatase （ALP） staining and quantitatively detect the activities of ALP in the BMSCs in various groups； On the 21th day of osteogenesis induction， the formations of calcium nodules in the BMSCs in various groups were detected by Alizarin red staining and the mineralization activities of cells in various groups were quantitatively detected； On the 7th day of osteogenesis induction， real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of ALP runt-related transcription factor 2 （Runx-2）， type Ⅰ collagen（COL-1），and osteocalcin （OCN） mRNA in cells in various groups； the immunofluorescence intensities of Runx-2 and OCN protein expressions in the BMSCs in various groups were measured by immunofluorescence staining. Results On the 1st and 3rd days of PPD treatment， compared with control group， the proliferation activity of the BMSCs in 40.0 mg·L^－1PPD group was significantly decreased （P<0.01）； on the 7th day of PPD treatment， compared with control group， there were no significant differences in the proliferation activities of the cells between 2.5，5.0， and 10.0 mg·L^－1 PPD groups （P>0.05），the proliferation activities of the BMSCs in 20.0 and 40.0 mg·L^－1 PPD groups were significantly decreased （P<0.01）. On the 1st and 3rd days of osteogensis induction，the numbers of alive cells in 5.0，10.0，and 20.0 mg·L^－1 PPD groups were more，so 10.0 mg·L^－1 PPD was selected to use for the following experiment.On the 7th day of osteogenesis induction，compared with control group， the activity of ALP in the cells in PPD group was significantly increased （P<0.01）； on the 21st day of osteogenesis induction， compared with control group， the number of mineralized nodules in the cells in PPD group was significantly increased，and the minealized activity was increased （P<0.01）； on the 7th day of osteogenesis induction， compared with control group， the expression levels of ALP，COL-1，OCN，and Runx-2 mRNA in the cells in PPD group were significantly increased（P<0.05），the fluorescence intensities of expressions of Runx-2 and OCN proteins in the cells in PPD group were significantly increased （P<0.01）. Conclusion PPD can promote the osteogenic induction of the BMSCs by increasing the ALP activity and calcium salt deposition in the BMSCs and increasing the expressions of osteogenic genes such as ALP，COL-1，OCN and Runx-2，and PPD is an effective osteogenesis induction activity factor.

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 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 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 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 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 screen the active ingredients of traditional Chinese medicine Sanghuang in the treatment of pneumonia based on network pharmacology， and to analyze the active ingredients and targets of Sanghuang in the treatment of pneumonia. Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） software was used to search for the ingredients and targets of Sanghuang； the Gene Name was obtained from the Uniprot database； the targets corresponding to pneumonia were found through GeneCards， and the targets of Sanghuang and pneumonia were compared to obtain the key targets； Cytoscape 3.9.1 was used to draw the chemical ingredients of Sanghuang-pneumonia-target network diagram； the protein-protein interaction （PPI） network diagram was constructed by String platform； Gene Ontology （GO） biological function analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway enrichment analysis on key genes of Sanghuang were performed by using the ClueGo plugin in Cytoscape software. Results There were 21 kinds of active ingredients in the traditional Chinese medicine Sanghuang， and （E）-4-（3，4-dihydroxyphenyl）but-3-en-2-one contained the most targets， including cytochrome P450 family （CYP450）， human epidermal growth factor receptor （EGFR）， and matrix metalloproteinase （MMP） and so on. A total of 358 targets were screened as the compound targets of Sanghuang，among them 217 genes were regarded as common targets for Sanghuang and pneumonia， and 69 key genes related to pneumonia were amine oxidase copper containing 3 gene （AOC3）， DNA ligase 1 gene （LIG1）， glutamyl aminopeptidase gene （ENPEP）， aldehyde dehydrogenase 2 family member gene （ALDH2）， PHD finger protein 8 gene （PHF8）， solute carrier family 11 member 2 gene （SLC11A2），CYP50 and so on.The KEGG and GO analysis results showed that Sanghuang mainly produced a marked effect through 17 metabolic pathways， including the phosphatidylinositol-3 kinase/protein kinase B （PI3K/Akt） signaling pathway， microRNAs（miRNA） in cancer， chemical carcinogenesis - receptor activation， prostate cancer， proteoglycans and lipids in cancer， atherosclerosis， chemical carcinogenesis-reactive oxygen species （ROS） and other related pathways. Conclusion （E）-4-（3，4-dihydroxyphenyl）but-3-en-2-one is the most effective chemical substance in the treatment of pneumonia among the ingredients of Sanghuang， and its mechanism is mainly related to the CYP450 family.

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 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 effect of microRNA-7 （miR-7） on the steroid-induced avascular nectosis of femoral head （SANFH） cell model in vitro，and to clarify its mechanism. Methods The lentivirus miR-7 mimic（miR-7 mimic） and miR negative control（miR NC） were transfected into the osteoblast MC3T3-E1 cells，and control group was set up， real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the transfection of MC3T3-E1 cells in various groups.The MC3T3-E1 cells were divided into control group， SANFH group， miR-7 mimic group， and miR-7 mimic+tumicamy （TM） group.CCK- 8 method was used to detect the proliferation activities of cells in various groups； flow cytometry was used to detect the apoptotic rates of cells in various groups； alkaline phosphatase （ALP） staining method was used to detect the ALP staining of cells in various groups；colorimetric method was used to detect the levels of ALP in cells in various groups； Alizarin red S staining was used to observe the formation of mineralization nodule in the cells in various groups； RT-qPCR method and Western blotting method were used to detect the expression levels of runt-related transcription factor 2 （Runx2）， osteocalcin （OCN）， osteopontin （OPN） mRNA and proteins in the cells in various groups； Western blotting method was used to detect the expression levels of glucose regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein（CHOP）， cysteinyl aspartate specific proteinase（Caspase）-12， Caspase-9， and Caspase-3 proteins in the cells in various groups； immunofluorescence double-labeling staining was used to detect the fluorescence intensities of expressions of GRP78 and Caspase-3 proteins in the cells in various groups. Results Compared with control group and miR NC group， the expression level of miR-7 in the MC3T3-E1 cells in miR-7 mimic group was increased （P<0.05）. Compared with control group， the proliferation activity of the MC3T3-E1 cells in SANFH group was decreased （P<0.05）， the apoptotic rate and TUNEL positive rate were increased （P<0.05）， the number of ALP staining cells was less and the ALP activity was decreased （P<0.05）， the number of formation of mineralized nodules in the cells was decreased， the expression levels of Runx2， OCN， and OPN mRNA and proteins in the cells were decreased （P<0.05）， the expressions levels of GRP78， CHOP， Caspase-12， Caspase-9， and Caspase-3 proteins in the cells were increased （P<0.05）， and the fluorescence intensities of expressions of GRP78 and Caspase-3 proteins were increased （P<0.05）； compared with SANFH group， the proliferation activity of the MC3T3-E1 cells in miR-7 mimic group was increased （P<0.05）， the apoptotic rate and TUNEL positive rate were decreased （P<0.05）， the number of ALP staining cells was increased， the ALP activity was increased （P<0.05）， the number of formation of mineralized nodules in the cells was significantly increased， the expressions levels of Runx2， OCN， and OPN mRNA and proteins in the cells were increased （P<0.05）， while the expression levels of GRP78， CHOP， Caspase-12， Caspase-9， and Caspase-3 proteins in the cells were decreased （P<0.05），and the fluorescence intensities of expressions of GRP78 and Caspase-3 proteins in the cells were decreased （P<0.05）；compared with miR-7 mimic group， the proliferation activity of the MC3T3-E1 cells in miR-7 mimic+TM group was decreased （P<0.05）， the apoptotic rate and TUNEL positive rate were increased （P<0.05）， the number of ALP staining cells was less and the ALP activity was decreased （P<0.05）， the number of formation of mineralized nodules in the cells was decreased， the expression levels of Runx2， OCN， and OPN mRNA and proteins in the cells were decreased （P<0.05）， the expression levels of GRP78， CHOP， Caspase-12， Caspase-9， and Caspase-3 proteins in the cells were increased （P<0.05）， and the fluorescence intensities of expressions of GRP78 and Caspase-3 proteins were increased （P<0.05）. Conclusion miR-7 can promote the osteogenesis of the MC3T3-E1 cells in SANFH state， and the mechanism may be related to the inhibition of endoplasmic reticulum stress-mediated apoptosis pathway.

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 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 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 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 effect of over-expression of Dectin-1 gene on the function of the dendritic cells （DCs）， and to clarify the mechanism of immune function of Dectin-1 gene in inhibiting the maturation and activation of DCs and its immune protection on the heart allografts. Methods The bone marrow stem cells of the mice were induced to form DCs and then cultured and expanded in vitro. The Dectin-1 gene was transfected into the DCs by adenovirus vector with green fluorescent protein （GFP）， the immature DCs （imDCs） modified with the Dectin-1 gene were regarded as DC-Dectin-1 group， and DCs group （untransfected with virus）and DC-GFP group（transfected with GFP） were sep up.Immunofluorescence assay used to detect the adenovirus transfection after 24 h； Western blotting method was used to detect the expressions of Dectin-1 protein in DCs in various groups；flow cytometry and enzyme-linked immunosorbent assay （ELISA） were used to detect the phenotype， function， and levels of interleukin-10 （IL-10） and interleukin-12 （IL-12） in cell culture supernatant of DCs in various groups before and after lipopolysaccharide（LPS） stimulation.The model of abdominal heterotopic heart allograft in the allogeneic mice was established， which was divided into DCs group， DC-GFP group，and DC-Dectin-1 group. The DCs， DC-GFP，and DC-Dectin-1 were infused on the first， third and fifth days after transplantation， respectively.On the seventh day after transplantation， HE staining was used to observe the pathomorphology of heart allografts of the mice in various groups； TUNEL staining was used to observe the apoptosis of heart allografts of the mice in various groups；the median survival time （MST） of heart allografts of mice in various groups was observed after transplantation. Results The gene modified Ad5F35 vector could improve the transfection efficiency of Dectin-1 gene， and GFP could continue to be expressed in the DCs after transfection. Before LPS stimulation， the expression levels of CD40， CD80， CD86， and human major histocompatibility complex Ⅱ （MHC-Ⅱ） in the cells in DCs， DC-GFP and DC-Dectin-1 groups were low， which presenting the imDCs phenotype； compared with before LPS stimulation， the expression levels of CD40， CD80， CD86， and MHC-Ⅱ in the cells in DCs and DC-GFP groups were increased after LPS stimulation， which presenting the mature DC phenotype. However， the expression levels of CD40， CD80， CD86， and MHC-Ⅱ in the cells in DC-Dectin-1 group were low. Before LPS stimulation， there were no statistically significant differences in the levels of IL-10 and IL-12 in cell culture supernatant in various groups（P>0.05）； after LPS stimulation， compared with DCs and DC-GFP groups， the IL-10 level in the cell culture supernatant in DC-Dectin-1 group was significantly increased （P<0.05）， while the IL-12 level was significantly decreased （P<0.05）. On the seventh day after transplantation， compared with DCs group and DC-GFP group， the inflammatory cell infiltration in heart allografts tissue of the mice in DC-Dectin-1 group was decreased， the rejection was significantly alleviated， and the number of TUNEL positive apoptotic cells were significantly decreased. Compared with DCs and DC-GFP groups， the MST of heart allografts of the mice in DC-Dectin-1 group was significantly prolonged （P<0.01）. Conclusion Dectin-1 gene can inhibit the maturation and activation of the immature DCs under the action of LPS， weaken their antigen presenting function， prolong the survival time of heart allografts of the mice to a certain extent， and induce the formation of allografts immune tolerance.

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 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.

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 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 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 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 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 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 optimal preparation process and the in vitro release properties of the poly （lactic-co-glycolic acid）poly（ethylene glycol）（PEG-PLGA） nanoparticles of triamcinolone acetonide（TA） and mycophenolate mofetil（MMF） in the treatment of thyroid associated ophthalmopathy（TAO）， and to evaluate its safety in the treatment of TAO by periorbital injection. Methods The TA nanoparticles （TA NPs group） and MMF nanoparticles （MMF NPs group） were prepared with PEG-PLGA as the raw material by emulsification method. The process optimization was carried out with the encapsulation efficiency as the evaluation indicator. The morphology of nanoparticles in various groups was observed under transmission electron microscope；the sizes of nanoparticles in various groups were detected by zetasizer particle size and potential analyzer. The release properties of nanoparticles in various groups were detected by ultraviolet spectrophotometry in vitro and the release rates of the drug were calculated，and then the release properties of the drugs were analyzed on the basis of clinical medication rules. The human retinal pigment epithelial hRPF-19 cells were divided into blank control group，different concentrations of TA groups，different concentrations of MMF groups，different concentrations of TA NPs group， and different concentrations of MMF NPs groups.The viabilities of the cells were evaluated by MTT test and the safty of preparation was evaluated. Results The TA-loaded and MMF-loaded nanoparticles were prepared （TA NPs and MMF NPs）. The encapsulation efficiencies of TA NPs and MMF NPs were 47.66% and 16.52%， with an average particle size of 600 nm. The potential of the nanoparticles met the basic requirements of periorbital injection. Under the microscope， the TA NPs and MMF NPs showed round appearance and a high degree of uniformity. The detection results of drug release system in vitro showed that the release characteristics of TA-NPs and MMF-NPs met the administration characteristics of TAO treatment drugs and the sustained release was able to last more than 3 weeks. The initial release rate was low and the release curve was stable. The MTT results showed that there were no significant cell inhibitions in different concentrations of TA groups， MMF groups， TA NPs groups，and MMF NPs groups at the lower concentration. At the higher concentration，compared with same concentration of TA group， the viabilities of the cells in 40，80，and 160 nmol·L^－1 TA NPs groups were increased （P<0.01）； compared with same concentration of MMF group，the viabilities of the cells in 50，100，and 200 nmol·L^－1 MMF NPs groups were increased （P<0.01）. Conclusion The physicochemical parameters of TA NPs and MMF NPs meet the basic requirements for the periorbital injection with simple preparation process and good safety and have better drug-loaded and sustained release effects， which can be consistent with the requirements for clinical drug therapy in the treatment of TAO.

Objective To prepare the strontium and nitrogen co-doped titanium dioxide （Sr-N-TiO₂）/nano hydroxyapatite （n-HA） composite resins， to evaluate its basic properties， antibacterial properties， remineralization abilities， and biosafeties， and to clarify the antibacterial mechanism of the new type of composite resins. Methods Sr-N-TiO₂ and n-HA were mixed as the reinforcing fillers. The composite resins were prepared and were divided into control group and three experimental groups. According to the mass fractions of reinforcing fillers， the experimental groups were divided into 2.5% reinforcing filler group， 5.0% reinforcing filler group， and 7.5% reinforcing filler group. The infrared spectrogram of composite resins in various groups were detected by the Fourier-transform infrared absorption spectrogram and the double bond conversion rates were calculated before and after light curing for 60 s. The curing depths of composite resin specimens in various groups after light curing for 20 s were calculated by cylindrical mould （height=10.0 mm， diameter=4.0 mm）. Three composite resin specimens in various groups were prepared and co-cultured with the dilute Streptococcus mutans solution. The plate colony counting method was used to determine the counts of attached bacteria on surface of the specimens and the antibacterial rates were calculated； live/dead bacteria staining method was used to observe the ratio of live bacteria/dead bacteria and morphology of composite resin speciments in various groups.The composite resin specimens were immersed in the artificial saliva for 1， 3， 5， and 7 d， and the mineralizations of surface of the composite resins were observed under scanning electron microscope （SEM）. The mouse fibroblast cells（L-929 cells） were incubated with the resin extract solution，and CCK-8 method was used to detect the relative growth rates （RGR） of the cells on the 1st， 2nd， and 3rd days and the cytotoxicity levels were evaluated. Results With the increasing of reinforcing filler proportion of the composite resins， the double bond conversion rates and curing depths of new type of the composite resins showed a decreasing trend， but all reached the clinical standard. Compared with control group， the double bond conversion rates of new type of the composite resins in 2.5% reinforcing filler group and 5.0% reinforcing filler group had no significant differences （P>0.05）， and the above index in 7.5% reinforcing filler group was decreased（P<0.05）. Compared with control group， the curing depths of the composite resins in experimental groups were increased （P<0.05 or P<0.01）， the counts of attached bacteria on surface of the speciments in experimental groups were significant increased （P<0.01）. When the proportion of reinforcing fillers reached 5.0%， the antibacterical rates of new type of the composite resins were higher than 90%. Compared with control group， the counts of live bacteria on surface of the composite resins in experimental groups were decreased with the increasing of proportion of the reinforcing fillers. The SEM results show that the mineralized nodules on surface of the composite resins in experimental groups could be observed， and the number of mineralized nodules was increased with the prolongation of time and the increasing of reinforcing fillers proportion. After treated for 7 d， the surface of the composite resins in 5.0% reinforcing filler group and 7.5% reinforcing filler group were almostly covered by mineralized nodules. The biosafety experiment results showed that the RGR of cells in various groups was above 75%，and the cytotoxicity level ≤1 grade. Conclusion The modified new type of composite resins meet the clinical standards， and it has antibacterial and remineralization properties， as well as high biosafety.