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

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

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

Objective To discuss the effect of G protein-coupled receptor kinase5（GRK5） over-expression on the depression-like behaviors of the P301S Tau transgenic mice，and to clarify its possible mechanism. Methods Seventeen male P301S Tau mice were randomly divided into blank control group （given no treatment） and negative control group （given negative control virus in bilateral hippocampus regions），over-expression group （given GRK5 over-expression virus in bilateral hippocampus regions）. The sugar-water preference test was used to detect the sugar-water preference rates of the mice in various groups， the percentages of immobility time of the mice in various groups were detected by tail suspension test（TST） and forced swimming test（FST）； the expression levels of neurological function-related proteins in hippocampus tissue of the mice in various groups were dectated by Western blotting method； the expressions of microglia marker protein （IBA1）， neuronal marker protein （NeuN）， and astrocyte marker protein （GFAP） in hippocampus tissue of the mice in various groups were detected by immunofluorescence staining. Result The sugar-water preference assay results showed that compared with blank control and negative control groups， the percentage of sugar-water preference of the mice in over-expression group was decreased（P<0.05）. The TST and FST results showed that the percentages of immobility time of the mice in over-expression group were increased compared with blank control and negative control groups （P<0.05）. The Western blotting results showed that the expression levels of GRK5， Tau T205， and IBA1 proteins in hippocampus tissue of the mice in over-expression group were increased （P<0.05） and the expression level of synatotagmin（SYN） protein was decreased （P<0.05）.The immunofluorescence staining results showed that compared with blank control group， the fluorescence expression of enhanced green fluoorescence protein（EGFP） in hippocampus tissue of the mice in negative control and over-expression groups could be seen and the EGFP was mainly co-localized with the NeuN； EGFP was hardly found in the microglia and astrocytes；compared with blank control and negative control groups， the number of microglia in hippocampus tissue of the mice in over-expression group was increased（P<0.05）. Conclusion Over-expression of GRK5 in hippocampus tissue of the P301S Tau mice induces the depressive-like behaviors，and its mechanism may be related to the increasing of the expression of GRK5 in nucleus of the neurons.

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

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

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

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

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

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

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

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

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

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

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

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

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

Objective To discuss the expression of circs_EFCAB2 in epileptic cell model in vitro，and to clarify its possible mechanism. Methods The magnesium-free epilepsy cell model （model group） was constructed in the human neuroblastoma LA-N-5 cells， and the cells untreated with magnesium-free extracellular fluid were regarded as control group. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of circ_EFCAB2 mRNA in the cells in two groups.The cells were didived into ribonuclease（RNase） R digestion group and RNase R undigestion group. RNA enzyme digestion experiment and RT-qPCR method were used to detect the expression levels of circ_ EFCAB2 and linear EFCAB2 mRNA in the cells in two groups； nuclear cytoplasmic separation experiment was used to detect the expression levels of circle_EFCAB2 mRNA in the epileptic cells； CCK-8 method was used to detect the proliferation activities of the cells in two groups； flow cytometry was used to detect the apoptotic rate and percentages of the cells at different cell cycles in two groups. Results The cells in model group showed spontaneous high-frequency epileptiform discharges， indicating the epilepsy cell model was successfully established. The RT-qPCR results showed that compared with control group， the expression level of circ_ EFCAB2 mRNA in the cells in model group was increased （P<0.05）. The RNA enzyme digestion experiment and RT-qPCR results showed that compared with RNase R undigestion group， the expression level of linear EFCAB2 mRNA in the cells in RNase R digestion group was decreased （P<0.01）. The nuclear cytoplasmic separation experiment results showed that there was no statistically significant difference in the expression level of circ_ EFCAB2 mRNA in the epileptic cells between the cytoplasm and nucleus of the epileptic cells（P>0.05）. The CCK-8 results showed that at 72 h after transfection， compared with control group， the proliferation activity of the cells in model group was significantly decreased （P<0.01）. The flow cytometry analysis results showed that compared with control group， the apoptotic rate of the cells in model group was significantly increased （P<0.01）；compared with control group， the percentage of the cells at S phase in model group was significantly decreased （P<0.01）， and the percentage of the cells at G₁+G₂ phase was significantly increased （P<0.01）. Conclusion The upregulated circ_ EFCAB2 may play a role in the pathogenesis of epilepsy by inhibiting the cell proliferation， promoting the apoptosis， and blocking the cell cycles.

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

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

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

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

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

Objective To discuss the influencing factors of oocytes retrieved numbers and oocytes retrieved rates during in vitro fertilization cycle in patients underwent in vitro fertilization/intracytoplasmic sperm injection（（IVF/ICSI）， and to provide the basis for the choice of the trigger time in clinical practice. Methods A total of 950 patients who received IVF/ICSI treatment were selected. The optimal scale regression model was used to analyze the effects of age， estradiol （E2） levels on the day of of human chorionic gonadotropin （HCG） administration， ovarian reserve function， percentages of different sizes of follicles on the days of oocyte retrieved and HCG administration， ovulation induction protocols of the patients， and operators on the number of oocyte retrieved and oocyte retrieved rate. Results There was a negative correlation between the age of patients and the number of oocytes retrieved（β=－0.068，P<0.01）； there was a negative correlation between the percentage of big follicles on the oocytes retrieved day and the number of oocyte retrieved（β=－0.243， P<0.01）， there was a negative correlation between the percentage of big follicles on the HCG administration day and the number of oocyte retrieved （β=－0.073，P<0.01）； there was a positive correlation between E2 levels on the HCG administration day and ovarian reserve function and the number of oocyte retrieved（β= 0.270，P<0.01；β=0.196， P<0.01）； there was a positive correlation between the percentage of middle follicles on the HCG administration day and the number of oocyte retrieved（β=0.098，P<0.01）.The numbers of oocyte retrieved of the patients in long protocol， antagonist protocol，progesterone-primed ovarian stimulation（PPOS） protocol， and natural cycle protocol were gradually decreased（β=0.227，P<0.01）. The regression analysis results showed that there were positive correlations between the percentages of big and middle follicles on the oocyte retrieved day and the rate of oocyte retrieved（β= 0.168， P<0.01；β=0.194， P<0.01）， the ovarian reserve function， ovulation induction protocol and different operators influenced the rate of oocyte retrieved（β=0.086，P=0.040；β=0.137，P<0.01； β=0.270，P<0.01）. Conclusion The percentage of follicles with diameter ≥ 18 mm on the HCG administration day should be controlled below 20%， and more oocytes should be retrieved； with the decreasing of the women’ age and ovarian reserve functions， the number of oocyte retrieved is gradually decreased； it is necessary to strengthen the standard procedures for oocyte retrieved surgery to promote the rate of oocyte retrieved.

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

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

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

Objectives To discuss the application of 3D printing auricular model in auricle reconstruction surgery and its effect on the symmetry and accuracy of the reconstructed auricles，and to provide the basis for evaluation on the clinical application of 3D auricular model. Methods A retrospective analysis was conducted on the clinical data of 76 patients with unilateral microtia treated with auricular reconstruction surgery.The normal auricle was scanned by the pre-operative CT to obtain the DICOM files， which were transformed into the light cured stereolithography （STL） files after mirroring and smoothing treatment. After being input into the 3D printer， the 3D auricular model which was consistent with the shape of normal auricle was printed. All the patients underwent the auricle reconstruction surgery by Nagata method. The first phase of the surgery was performed by using the costal cartilage as the material， the auricle scaffold was reconstructed and fine carved according to the 3D printing auricular model； the second phase of the surgery was enhancing the reconstructed auricle and forming the otocranial sulcus according to the 3D printing auricular model. The postoperative follow-up time was 6－12 months， and the length and width of the reconstructed and normal auricles，the length， width and depth of the conchal cavity， depth of the triangular fossa， and the cranioauricular angle were measured and compared. The satisfaction degrees of the patients with the position， size， cranioauricular angle， symmetry， and stability of the reconstructed auricles were investigated， and the satisfaction rates of the patient were calculated. Results A total of 76 patients received postoperative follow-up and all the reconstructed auricles were survived. The satisfaction rates of the patients and their families for the symmetry of the reconstructed auricle reached 85.5%， and the satisfaction rates for the size， position， and stability were all > 90%， the satisfaction rate for the otocranial angle was 68.4%， and the average satisfaction rate was 87.4%. Compared with healthy side auricle， there were no statistically significant differences in the length and width of the reconstructed auricle， the length and depth of the conchal cavity， and the depth of the triangular fossa （P>0.05）. Compared with healthy side auricle，the otocranial angle of the reconstructed auricle was significantly decreased（P=0.014 1）. The similarity of shape between the reconstructed auricle and the healthy side auricle was high， and the similarity of fine structure of the auricle was also high. Conclusion The 3D printing auricular models based on the 3D printing technology can make the reconstructed auricle to be similar highly with the healthy side auricle， and it is worth to apply in the clinical treatment.

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

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

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

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