Objective To discuss the potential targets and signaling pathways of Tonifying Slpeen and Kidney Empirical Prescription （TSKEP） in the treatment of neuromyelitis optica spectrum disorders（NMOSD） through network pharmacology analysis and animal experiment，and to provide the theoretical basis for the treatment of NMOSD. Method Traditional Chinese Medicine System Analysis Platform（TCMSP） Database was used to screen the chemical compounds and targets of TSKEP； the related target genes in NMOSD were comfirmed through GendCard and other Databases；the core compounds were obtained by building the compound-target network； the STRING Database and the Network Analyzer plugin in Cytoscape 3.6.0 software were used to obtain the topology parameters of protein-protein interaction （PPI） network and to identify the core targets； Omicshare platform was used to perform the Geno Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis and the target-pathway network was conducted to identify core key biological processes and signaling pathways； the molecular docking of core compounds and core targets was performed based on AutoDock software. Eighteen mice were divided into control group， NMOSD group， and TSKEP group， and there were 6 mice in each group. The morphology of brain tissue of the mice in various groups were observed by HE staining，and the levels of related cytokines in serum of the mice were detected by enzyme-linked immunosorbent assay（ELISA） method. Results The animal experiment results showed that compared with NMOSD group，the score of neural function defect of the mice in TSKEP group was decreased（P<0.01）， and the degree of inflammatory cell infiltration in central nervous system of the mice in TSKEP group was allevited. A total of 304 active compounds and 655 drug targets were screened， and 43 potential targets were related to the treatment of NMOSD with TSKEP. Quercetin， luteolin， and stigmasterol may be the core compounds，and interleukin-6（IL-6），interlleukin-1β（IL-1β），tumor necrosis factor-α（TNF-α） could be the poetential core targets.The KEGG pathway enrichment analysis and network analysis results showed that TSKEP may play an important role in the treatment of NMOSD through 14 signaling pathways.The molecular docking verification results showed that core compounds combined well with the core targets. Compared with NMOSD group and control group， the levels of serum inflammatory cytokines of the mice in TSKEP group were decreased（P<0.05 or P<0.01）. Conclusion Based on the verification of network pharmacology and animal experiment， TSKEP can exert the therapeutic effect through regulating the expressions of serum inflammatory cytokines， promoting the repairment and regeneration of nerve function，and reducing the inflammatory response of the central nervous system by multiple targets and signaling pathways.

Objective To investigate the effects of translocator protein （TSPO） ligand XBD173 on the pulmonary inflammatory response and M1 polarization of macrophages of the mice induced by cigarette smoke extraction （CSE）， and to clarify their realted molecular mechanisms. Methods A total of 18 adult male C57BL/6 mice were randomly divided into control group， CSE group，and CSE+XBD173 group.The mice in CSE group were intranasally given 20 μL CSE，and the mice in CSE+XBD173 group were given the same dose of CSE and intraperitoneally injectied with 10 mg·kg^－1 XBD173. The pathomorphology of lung tissue of the mice in various groups was observed by HE staining and PAS staining. The RAW264.7 macrophage were cultured and divided into control group， CSE group， and CSE + XBD173 group. The CSE administration concentration was 1%， the XBD173 administration concentration was 4 mg·L^－1， and the cells in control group were given the same volume of dimethyl sulfoxide；after 18 h of stimulation， the expression levels of CD80， CD86， inducible nitric oxide synthase （iNOS）， reactive oxygen species （ROS） in the RAW264.7 cells，and the apoptotic rates of the RAW264.7 cells in various groups were detected by flow cytometry； Western blotting method was used to detect the expression levels of nuclear factor-κB/p65 （NF-κB/p65） and phosphorylated NF-κB/p65 （p-NF-κB/p65）proteins in the cells in various groups. The siRNA was used to knock down the TSPO expression in the RAW264.7 cells in various groups， and the expression levels of interleukin-6 （IL-6） and tumor necrosis factor-α（TNF-α） mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. Results The animal experiment results showed that compared with control group， the number of inflammatory cells in lung tissue of the mice in CSE group was increased significantly and the bronchial wall was thicker；compared with CSE group， the number of inflammatory cells in lung tissue of the mice in XBD173+CSE group was decreased and the bronchial wall was thinner. The cell experiment results showed that compared with control group， the expression levels of CD80， CD86， iNOS， and ROS in the RAW264.7 cells in CSE group were increased（P<0.05）， the apoptotic rate was increased（P<0.05）， the expression levels of IL-6 and TNF-α mRNA were increased（P<0.05），and the expression level of p-NF-κB/p65 protein was significantly increased（P<0.05）， while the expression level of NF-κB/p65 had no significant difference （P>0.05）； compared with CSE group， the expression levels of CD80， CD86， iNOS， and ROS in the RAW264.7 cells in CSE+XBD173 group were decreased（P<0.05）， the apoptotic rate was decreased（P<0.05）， the expression levels of IL-6 and TNF-α mRNA were decreased（P<0.05）， and the expression level of p-NF-κB/p65 protein was decreased significantly（P<0.05）. After the expression of TSPO protein in the RAW264.7 cells was knocked down by siRNA， the expression levels of IL-6 and TNF-α mRNA in the RAW264.7 cells among various groups had no significant differences（P>0.05）. Conclusion XBD173 can alleviate the inflammatory response of lung tissue of the mice and inhibit the M1 polarization of the macrophage induced by CSE， and its mechanism may be related to the NF-κB protein.

Objective To investigate the improvement effect of effective-compound compatibility of Bufei Yishen prescrtion（ECC-BYP Ⅲ） on pulmonary hypertension （PH） of the rats induced by smoke and bacteria， and to preliminarily elucidate its mechanism. Methods SPF SD rats were randomly divided into control group （n=9） and modeling group（n=140）.The rats in control groujp were given conventional feeding，and the rats in modeling group were given cigarette-smoke and Klebsiellabacillus （Kp） to make the PH model. Then the rats in model group were divided into model group（n=10）， low（n=9）， medium（n=10）， and high（n=10） doses （3.24， 6.48 and 12.96 mg·kg^－1·d^－1） of ECC-BYP Ⅲgroups， according to the equivalent pulmonary function of the rats among various groups.The rats in ECC-BYP Ⅲ groups were administered with different doses of ECC-BYP Ⅲ， respectively， and the rats in control and model groups were administered with same volume of solvent for 4 weeks. On the 29th day，the mean pulmonary artery pressure （mPAP）， pulmonary artery systolic pressure （PASP），pulmonary artery diastolic pressure （PADP）， and right ventricular hypertrophy index （RVHI） of the rats in various groups were measured.The lung tissues of the rats in various groups were taken out. Hematoxylin-eosin（HE）staining was used to detect the periarteriolar inflammation in lung tissue；victoria blue staining was used to detect the percentages of non-muscular， partial muscular， and muscular pulmonary arterioles in pulmonary arterioles of the rats， the percentages of wall thickness of pulmonary muscular arterioles in vessel diameter（WT%），and luminal area in pulmonary muscular arterioles （LA%）. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of interleukin 6 （IL-6）， tumor necrosis factor （TNF-α）， endothelin-1 （ET-1）， and protracyclin （PGI2） in lung tissue of the rats in various groups. Results Compared with control group，the mPAP， PASP， PADP of the rats in model group were increased（P<0.05 or P<0.01）， RVHI was increased（P<0.01），and there were obvious inflammatory cell infiltration around the pulmonary arterioles； the levels of TNF-α and IL-6 in lung tissue were increased（P<0.01），the percentage of muscular arteriole and WT% of pulmonary were increased and non-muscular arteriole and LA% were decreased（P<0.05 or P<0.01）， and the level of ET-1 and ET-1/PGI2 ratio in lung tissue was increased（P<0.01）. Compared with model group， the mPAP and PADP of the rats in high dose of ECC-BYP Ⅲ group were decreased（P<0.05），the inflammatory cell infiltration of the rats in medium and high doses of ECC-BYP Ⅲ groups was improved，and the levels of TNF-α， IL-6， WT%，percentage of muscular arteriole were decreased（P<0.01），and LA% was increased （P<0.01）；compared with model group，the ET-1 levels in lung tissue of the rats in medium and high dose of ECC-BYP Ⅲ groups were decreased（P<0.05 or P<0.01），and the ET-1/PGI2 ratio of the rats in high of ECC-BYP Ⅲ group was decreased（P<0.05）.Compared with low dose of ECC-BYP Ⅲ group， the WT % and ET-1 in lung tissue were decreased（P<0.05 or P<0.01） and LA % was increased（P<0.01），the levels of TNF-α in lung tissue of the rats in medium dose of ECC-BYP Ⅲ group was decreased and the level of IL-6 and ET-1/PGI2 ratio in lung tissue of the rats in high dose of ECC-BYP Ⅲ group were decreased（P<0.05 ）. Conclusion ECC-BYP Ⅲ can reduce PH and wall thickness of the pulmonary arterioles， improve the lumen stenosis and pulmonary arteriolation，improve the pulmonary vascular remodeling，and its mechanism may be related to reducing the inflammation response of the pulmonary perivascular and improve unbalance of peripheral vasoconstriction vasular factor level/diastolic vascular factor level.

Objective To investigate the types， characteristics， and differences of microflora in various regions of the intestinal tract of the mice with chronic gastritisinfected with Helicobacter pylori （Hp）， and to clarify the related mechanism. Methods Thirty C57BL/6 mice were randomly divided into control group and infection group， and there were 15 mice in each group. The mice in control group were given normal saline， and the mice in infection group were given Hpsuspension with the concentration of 1×10⁹ CFU·mL^－1. After the model was successful builted， the contents of duodenum， jejunum， and colon of the mice in two groups were collected to extract the total DNA， and PCR amplification was performed； the 16SrRNAV3-V4 region was sequenced through high-throughput sequencing technique；the characteristics， differences，and diversities of the microflora in various regions of intestinal tract of the mice in control and infection group were analyzed by α and β diversity analysis. Results The total number of operational taxonomic unit（OTU） in two groups generated by cluster analysis samples was 211. At the phylum level， there were 5 dominant bacteria such as Firmicutes， Bacteroidetes， Actinomyces， proteobacteria， and Verrucomicrobia； compared with control group， the abundances of Actinobacteria in the duodenum， jejunum and colon of the mice in infection group were significantly decreased （P<0.05）.At the genus level，there were 21 dominant bacteria such as Lactobacillus， Staphylococcus， Clostridium Bacillus-ⅩⅣa， Turicibacter， Akkermansia， Bifidobacterium， Saccharibacteria-genera-incertae-sedis， Barnesia， Desulfovibrio， Alternaria， and Mycoplasma and so on； compared with control group， the abundances of Lactobacillus and Bifidobacterium in the duodenum and colon of the mice in infection group were significantly decreased （P<0.05）， and the abundance of Mycoplasma was significantly increased （P<0.05）.The OTU principal component analysis （PCA） results showed that there were no significant differences in the principal components between two groups（P>0.05）.In the Alpha diversity analysis，the shannon index analysis results showed that there were significant differences in the diversities of duodenum and jejunum microflora of the mice between control group and infection group（P<0.01），and the diversity of microflora in infection group was decreased； the simpson index analysis results showed that there were statistically significant differences in the diversities of duodenum， jejunum and colon microflora of the mice between control group and infection group （P<0.05）；and the diversity of microflora in infection group was decreased.The Welch’s t-test analysis results showed that the abundances in various regions of intestinal tract of the mice in two groups had signifricant differences（P<0.05）. Conclusion The types and characteristics of the microflora in various regions of intestinal tract of the mice with chronic gastritis infected with Hp change， the abundance of the probiotics is decreased and the abundance of Mycoplasma is increased. Its mechanism may be related with the changes of intestinal microenviroment，induced by the colonization of Hp.

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 17 （lncRNA SNHG17） on the biological behavior of the non-small cell lung cancer （NSCLC） cells， and to clarify its mechanism. Methods The human NSCLC A549 cells and H1299 cells were cultured，and the cells were transfected with pcDNA3.1-NC， SNHG17 over-expression plasmid（pcDNA3.1-SNHG17）， si-NC， SNHG17 small interference RNA（si-SNHG17），mimics NC，microRNA-384 mimics（miR-384 mimics）， inhibitor NC， miR-384 inhibitor（miR-384 inhibitor）， pcDNA3.1-astrocyte elevated gene 1 （AEG1）， and si-AEG1， respectively. Real-time quantitative PCR （RT-qPCR） method was used to detect the expression levels of lncRNA SNHG17，miR-384， and AEG1 mRNA in the A549 cells， H1299 cells and cells in various groups after transfection； ENCORI and TargetScan Databases were used to predicte the targeted bindings between lncRNA SNHG17 and miR-384，miR-384 and AEG1 mRNA. The A549 cells were divided into si-NC group， si-SNHG17 group， inhibitor NC group， miR-384 inhibitor group， si-NC+inhibitor NC group， si-SNHG17+inhibitor NC group， and si-SNHG17+miR-384 inhibitor group， si-AEG1 group， inhibitor NC+si-NC group， miR-384 inhibitor+si-NC group and miR-384 inhibitor+si-AEG1 group.The H1299 cells were divided into pcDNA3.1-NC group， pcDNA3.1-SNHG17 group， mimics NC group， miR-384 mimics group， pcDNA3.1-NC+mimics NC group， pcDNA3.1-SNHG17+mimics NC group，pcDNA3.1-SNHG17+miR-384 mimics group， pcDNA3.1-AEG1 group， mimics NC+pcDNA3.1-NC group， miR-384 mimics+pcDNA3.1-NC group， and miR-384 mimics+pcDNA3.1-AEG1 group. CCK-8 method was used to detect the cell viabilities in various groups； Transwell method was used to detect the numbers of migration and invasion cells in various groups； Western blotting method was used to detect the expression levels of AEG1 protein in the cells in various groups. Results The expression level of lncRNA SNHG17 in the H1299 cells was significantly lower than that in the A549 cells （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of SNHG17 in the cells in si-SNHG17 group was significantly decreased （P<0.01），the cell viability， number of migration cells， and number of invasion cells were significantly decreased （P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of SNHG17 in the cells in pcDNA3.1-SNHG17 group was significantly increased （P<0.01）， the cell viability，number of migration cells， and number of invasion cells were increased significantly （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of miR-384 in the cells in si-SNHG17 group was significantly increased（P<0.01）， and the expression level of AEG1 mRNA was significantly decreased（P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of miR-384 in the cells in pcDNA3.1-SNHG17 group was significantly decreased（P<0.01）， and the expression level of AEG1 mRNA was significantly increased （P<0.01）. The ENCORI Database results predicted that there were two binding sites between SNHG17 and miR-384. The rescue assay results showed that in the A549 cells， compared with si-NC+inhibitor NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in si-SNHG17+inhibitor NC group were significantly decreased（P<0.01）； compared with si-SNHG17+inhibitor NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in si-SNHG17+miR-384 inhibitor group were significantly increased（P<0.01）. In the H1299 cells， compared with pcDNA3.1-NC+mimics NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in pcDNA3.1-SNHG17+mimics NC group were significantly increased （P<0.01）； compared with pcDNA3.1-SNHG17+mimics NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in pcDNA3.1-SNHG17+miR-384 mimics group were significantly decreased （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of AEG1 protein in the cells， cell viability， number of migration cells， and number of invasion cells in si-AEG1 group were significantly decreased （P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of AEG1 protein cell viability， number of migration cells， and number of invasion cells were significantly increased （P<0.01）. The ENCORI Database results predicted that there was one binding site between miR-384 and AEG1.In the A549 cells， compared with inhibitor NC+si-NC group， the cell viability， number of migration cells and number of invasion cells in miR-384 inhibitor+si-NC group were significantly increased （P<0.01）； compared with miR-384 inhibitor+si-NC group， the cell viability， number of migration cells and number of invasion cells in miR-384 inhibitor+si-AEG1 group were significantly decreased （P<0.01）. In the H1299 cells， compared with the mimics NC+pcDNA3.1-NC group， the cell viability， number of migration cells， and number of invasion cells in miR-384 mimics+pcDNA3.1-NC group were significantly decreased （P<0.01）； compared with miR-384 mimics+pcDNA3.1-NC group， the cell viability， number of migration cells， and number of invasion cells in miR-384 mimics+pcDNA3.1-AEG1 group were significantly increased （P<0.01）. Conclusion LncRNA SNHG17 regulates the proliferation， migration， and invasion of the NSCLC cells by targeting AEG1 through miR-384.

objective To investigate the immunological characteristics of Acanthamoeba castellanii （Ac） Actin （Actin 1） （Ac-Action 1），and to clarify the role of Ac-Actin 1 in adhesion to the host cells and invasion of Ac parasite preliminarily. Methods The Ac trophozoite cDNA was regarded as the template， and the prokaryotic expression vector pET 22b（+）-Ac-Actin 1 was constructed and transformed into the Escherichia coli competent cells BL21 （DE3）；the recombinant Ac-Actin 1 protein （rAc-Actin 1） was induced by 1 mmol·L^－1 isopropyl β-D-thiogalactoside（IPTG） in vitro，and the solubility of rAc-Actin 1 was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）；the rAc-Actin 1 with His label was purified by affinity chromatography. Three New Zealand white rabbits were randomly divided into control group（n=1） and experiment group（n=2）； the rabbits in experiment group were subcutaneously injectied with 400 μg rAc-Actin 1 as the immunogen and the rabbit in control group was injected with the same amount of normal saline，and the anti-rAc-Actin 1 polyclonal serum of the rabbits was prepared；the titers of anti-rAc-Actin 1 polyclonal antibody of the rabbits in two groups were determined by enzyme linked immunosorbent assay（ELISA） method and the IgG subtype was determined； Western blotting method was used to detect the immunoreactivity of anti-rAc-Actin 1 polyclonal rabbit serum and rAc-Actin 1；the localization of Ac-Actin 1 in the Actrophozoite was detected by the indirect immunofluorescence assay （IFA）. The normal trophozoites were used as controls， the anti-rAc-Actin 1 polyclonal serum of the rabbits blocked the Ac trophozoites and was co-incubated with the Vero cells，and the adhesion effect of Ac-Actin 1 on the Vero cells was observed by microscope. Results The SDS-PAGE and BAC concentration detection results showed that the high concentration of rAc-Actin 1 （1.7 g·L^－1） protein was obtained.The ELISA results showed that the titer of the obtained rabbit anti-rAc-Actin 1 specific polyclonal antibody was 1∶6 400，and the concentrations of IgG1 and IgG2a were 116.76 g·L^－1 and 1 136.15 mg·L^－1， respectively.The Western blotting results showed that the rabbit anti-rAc-Actin 1 polyclonal antibody could specifically bind to the rAc-Actin 1 and had good immunoreactivity.The IFA results showed that rAc-Actin 1 mainly located on the cell membrane of Ac trophozoites.The microscope analysis results showed that compared with control group， the adhesion rate of Ac trophozoites to the Vero cells in the experiment group was significantly decreased（P<0.01） with the prolongation of incubation time of antibody and parasite， and the anti-rAc-Actin 1 specific polyclonal antibody could effectively block the adhesion of Ac trophozoites to the Vero cells. Conclusion rAc-Actin 1 has good immunogenicity and immunoreactivities， which is involved in the adhesion between the Ac parasite and the host cells.

Objective To investigate the effect of poiaic acid （PA） on the migration， invasion， and epithelial-mesenchymal transition（EMT） of the pancreatic cancer PANC-1 cells by up regulating the expressions of activating transcription factor 3 （ATF3） and heat shock protein family A member 6（HSPA6），and to clarify its possible mechanism. Methods The pancreatic cancer PANC-1 cells were divided into blank control group and different concentrations （2，5，10，20，30，40， and 50 μmol·L^－1）of PA groups，and the activities of the PANC-1 cells in various groups were detected by CCK-8 method. The PANC-1 cells were treated with different concentrations （0， 10， 30， and 50 μmol·L^－1） of PA， and the migration and invasion abilities of the PANC-1 cells were detected by Transwell chamber experiment， and the expression level of EMT-related proteins in the PANC-1 cells were detected by Western blotting method. A total of 10 BALB/c nude mice were randomly divided into control group and PA group， and there were 5 mice in each group. The PANC-1 cells were subcutaneously injected into the nude mice. When the tumor volume reached 60 mm³， the nude mice in PA group were intraperitoneally injected with 25 mg·kg^－1 PA， and the nude mice in control group were injected with the same volume of normal saline， the tumor volume and tumor weight of the mice in various groups were measured， and the expressions of Ki-67 in xenografted tissue of the mice in various groups were detected by immunohistochemistry. The differentially expressed genes of the PA-treated and PA-untreated pancreatic cancer cells in the GSE64111 Dataset were analyzed by GEO2R software. The PANC-1 cells were treated with different concentrations（0 and 30 μmol·L^－1） of PA， and the expression levels of HSPA6 and ATF3 proteins in the PANC-1 cells were detected by Western blotting method. The PANC-1 cells treated with 30 μmol·L^－1 PA were randomly divided into si-NC group and si-ATF3 group， and the cells were transfected with control siRNA and ATF3 siRNA， respectively. The expression levels of HSPA6 and ATF3 proteins and EMT-related proteins in the PANC-1 cells in various groups were detected by Western blotting method， and the migration and invasion abilities of the cells were detected by Transwell chamber experiment. Results The CCK-8 experiment results showed that compared with blank control group， the activities of the PANC-1 cells in different concentrations of PA groups were significantly decreased （P<0.05）.The results of Transwell chamber experiment showed that compared with blank control group， the migration and invasion abilities of the PANC-1 cells in different concentrations of PA groups were significantly decreased （P<0.05），and showed a concentration depent manner. The Western blotting results showed that compared with blank control group， the expression levels of E-cadherin protein in the PANC-1 cells in different concentrations of PA groups were significantly increased （P<0.05）， while the expression levels of N-cadherin and Vimentin proteins were significantly decreased （P<0.05）.The results of tumor formation in the nude mice showed that compared with control group， the volume and weight of the xenografted of the mice in PA group were significantly decreased （P<0.05）；the immunohistochemistry results showed that the Ki-67 staining in xeografted was shallow； the GEO2R software analysis and Western blotting results showed that compared with blank control group， the expression levels of HSPA6 and ATF3 proteins in the PANC-1 cells in PA group were significantly increased （P<0.05）. The Western blotting results showed that compared with si-NC group， the expression levels of ATF3， HSPA6， and E-cadherin proteins in the PANC-1 cells in si-ATF3 group were significantly decreased （P<0.05）， while the expression levels of N-cadherin and Vimentin proteins were significantly increased （P<0.05）.The Transwell chamber experiment results showed that compared with si-NC group， the migration and invasion abilities of the PANC-1 cells in si-ATF3 group were significantly increased （P<0.05）. Conclusion PA inhibits the migration， invasion， and EMT of the pancreatic cancer cells by up-regulating the expressions of HSPA6 and ATF3， thus play an anti-pancreatic cancer effect.

Objective To investigate the effects of atorvastatin （ATO） on the proliferation， apoptosis， and migration of the human tongue squamous cell carcinoma CAL-27 cells in vitro， and to clarify their mechanisms. Methods The CAL-27 cells were divided into control group and 1，5，10，20，and 40 μmol?L^－1 ATO groups.After treated with ATO，CCK-8 assay was used to detect the survival rates of the CAL-27 cells in various groups， clone formation assay was used to detect the clone formation rates of the CAL-27 cells in various groups； Hoechst33342 fluorescence staining and flow cytometry were used to detect the apoptotic rates of the CAL-27 cells in various groups； cell scratch test was used to detect the migration rates the CAL-27 cells in various groups； Western blotting method was used to detect the expression levels of P53， P21， B-cell lymphoma-2（Bcl-2），Bcl-2 associated X protein（Bax）， cysteinyl aspartate specific proteinase（Caspase）-3， Caspase-9， and cyclin-dependent kinase 6（CDK6） proteins in the CAL-27 cells in various groups. Results Compared with control group， the survival rates of the cells in different concentrations of ATO groups were decreased after cutured for 24 and 48 h（P<0.05）.After cultured for 48 h， compared with control group， the clone formation rates （P<0.01） and migration rates （P<0.05） of the cells in different concentrations of ATO groups were decreased，and the cells in 40 μmol?L^－1 ATO group basically lost the abilities of cloning and migration； the apoptotic rates in different concentrations of ATO groups were increased （P<0.05），and almost all the cells in 40 μmol?L^－1ATO group were apoptotic. After cultured for 48 h， compared with control group， the cells in different concentrations of ATO groups showed nuclear staining or fragmentation，and all the cells in 20 and 40 μmol?L^－1 ATO groups were fragmented. Compared with control group， after cultured for 48 h，the expression levels of P53， P21， Bax， Caspase-3， and Caspase-9 proteins in the cells in different concentrations of ATO groups were increased （P<0.01）， and the expression levels of CDK6 and Bcl-2 proteins were decreased（P<0.05 or P<0.01）. Conclusion ATO inhibits the proliferation and migration of the CAL-27 cells and promotes the apoptosis of the CAL-27 cells through P53 /P21 / CDK6 pathway.

Objective To observe the effect of curcumin on proliferation and invasion of the gastric cancer MGC-803 cells in vitro and in vivo， and to discuss the mechanism of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian rapamycin target protein （mTOR） signaling pathway. Methods The gastric cancer MGC-803 cells were divided into control group， low dose （10 μmol·L^－1） of curcumin group， middle dose （20 μmol·L^－1） of curcumin group，and high dose （40 μmol·L^－1） of curcumin group. The proliferation abilities of the MGC-803 cells in various groups were detected by CCK-8 method；the invasion abilities of the MGC-803 cells in various groups were detected by Transwell method；the apoptotic rates of the MGC-803 cells were detected by flow cytometry. The MGC-803 cells were used to establish the gastric cancer transplanted tumor model and the mice were treated with low（0.5 mg·kg^－1）， middle （1.0 mg·kg^－1），and high （2.0 mg·kg^－1） doses of curcumin， and the inhibitory effect of curcumin on the tumor growth of the mice in various groups was observed. TUNEL method was used to detect the apoptosis of the tumor cells in transplanted tumor tissue of the mice in various groups；the expression levels of PI3K-Akt-mTOR signaling pathway proteins in transplacted tumor tissue of the mice in various groups were detected by Western blotting ththod. Results Compared with control group， the proliferation abilities and number of invasion cells in different doses of curcumin groups were significantly decreased （P<0.05）， and the apoptotic rates were significantly increased （P<0.05）； the cell proliferation ability and the number of invasion cells in low dose of curcumin group were significantly higher than those in high dose of curcumin group （P<0.05）， and the apoptotic rate was significantly lower than that in high dose of curcumin group （P<0.05）.Compared with control group， the volumes of transplanted tumor of the mice in different doses of curcumin groups were significantly decreased （P<0.05）.The apoptotic rate of the cells in transplanted tumor tissue of the mice in control group was low， and the apoptotic rate of the cells in transplated tumor tissue of the mice in middle dose of curcumin group was significantly higher than that in low dose of curcumin group （P<0.05）， while the apoptotic rate of the cells in transplated tumor tissue of the mice in high dose of curcumin group was significantly higher than that in middle dose of curcumin group （P<0.05）. Compared with control group， the proteins expression levels of PI3K， phosphorylated Akt（p-Akt） and phosphorylated mTOR（p-mTOR） in transplanted tumor tissue of the mice in different doses of curcumin groups were significantly decreased （P<0.05）；the expression levels of PI3K， p-Akt， and p-mTOR proteins in transplanted tumor tissue of the mice in low dose of curcumin group were significantly lower than those in middle dose of curcumin group （P<0.05）； the expression levels of PI3K， p-Akt， and p-mTOR proteins in transplanted tumor tissue of the mice in high dose of curcumin group were significantly lower than those in middle dose of curcumin group （P<0.05）. Conclusion Curcumin can effectively inhibit the proliferation of tumor， decrease the invasion ability， and increase the apoptotic rate of the tumor cells；its mechanism may be related to down-regulation of expressions of PI3K/Akt/mTOR signaling pathway proteins.

Objective To screen the active components of Tongluo Tangtai Power（TLTT） in the treatment of diabetic peripheral neuropathy（DPN），predict the action targets and signaling pathways by network pharmacology，and to explore its mechanism. Methods The Traditional Chinese Medicine System Pharmacological Analysis Platform （TCMSP） was used to collect the main chemical components and action targets of TLTT；Genecard Database was used to retrieve the proteins associated with DPN； the protein-protein interaction（PPI） network was constructed by STRING Database and network visualization software Cytoscape3.9.1； pathway enrichment analysis and composition-target docking were analyzed by the Kyoto Encyclopedia of Genes and Genomes （KEGG） and molecular docking technique. Thirty SD rats were divided into control group， mecobalamin group， and TLTT group， and there were 10 rats in each group.The rats were given mecobalamin and TLTT by gavage to make the drug-containing serum； one stain of RSC96 Schwann cells was resuscitated；control group， high glucose model group， mecobalamin group， and TLTT group were set up；then cell counting method was used to detect the survival rates of Schwann cells in various groups； Western blotting method was used to detect the expression levels of serine/threonine kinase 1（AKT1） and tumor suppressor gene TP53 proteins in the cells in various groups. Results There were 95 active compound components and 463 targets in TLTT for the treatment of DPN， 1 187 DPN-related proteins， and 137 drug-disease common targets.The Geno Ontology（GO） analysis，including biological processes， cell composition，and molecular function） results obtained 5 505， 466， and 791 results；the KEGG analysis obtained 253 results，suggesting that TLTT may played a therapeutic role by regulating the phosphatidyl inositol 3 kinase/protein kinase B （PI3K-Akt） signaling pathway， liquid shear stress and atherosclerosis， advanced glycosylation end products and their receptors （AGE-RAGE），mitosolysis-activated protein kinase （MAPK） singling pathways and so on；the molecular docking results showed that TLTT acted on AKT1，TP53，tumor necrosis factor（TNF），epidermal growth factor receptor（EGFR），and other core targets by some core components such as quercetin，sterol，beta-sitosterol， kaempferol， ivy saponin，and so on. The CCK-8 results showed that compared with control group，the survival rate of the cells in high glucose model group was significantly decreased（P<0.05）；compared with high glucose model group，the viability of the cells in TLTT group was significantly increased （P<0.05）.The Western blotting results showed that compared with control group， the expression level of AKT1 the cells in high glucose model group was significantly decreased（P<0.05）， and the expression level of TP53 protein was significantly increased （P<0.05）；compared with high glucose model group， the expression level of AKT1 protein in the cells in TLTT group was significantly increased（P<0.05）， and the expression level of TP53 protein was significantly decreased（P<0.05）. Conclusion The main active components of TLTT， such as quercetin， may promote the survival of the Schwann cells， increase the expression level of AKT1 protein and decrease the expression level of TP53 protein by targeting PI3K-Akt and other signaling pathways， so as to play a role in the treatment of DPN.

Objective To investigate the inhibitory effect of rosiglitazone on ferroptosis of renal tubular epithelial cells in the mice with acute kidney injury （AKI） induced by lipopolysaccharide （LPS）， and to clarify its mechanism. Methods A total of 18 C57BL/6 male mice were randomly divided into control group（n=6）， LPS group（n=6），and LPS +rosiglitazone group （n=6）. The mice in LPS group and LPS+rosiglitazone group were injected with 10 mg·kg^－1 LPS intraperitoneally； the mice in LPS+rosiglitazone group were injected with 0.5 mg·kg^－1 rosiglitazone via the tail vein 30 min before LPS injection；the mice in control group were injected with the same volume of normal saline. The mice were sacrificed 24 h after LPS injection， and the kidney tissue and serum were collected. HE staining was used to observe the pathomorphology of kidney tissue of the mice in various groups； the levels of serum creatinine （CRE） and urea nitrogen （BUN） of the mice were detected by spectrophotometry； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of acyl CoA synthetase long-chain family member 4（ACSL4），glutathione peroxidase 4（GPX4）， solute carrier family 7 member 11（SLC7A11），interleukin-6（IL-6）， interleukin-1β（IL-1β）， and tumor necrosis factor-α（TNF-α） mRNA in kidney tissue of the mice in various groups；Western blotting method was used to detect the expression levels of ACSL4，GPX4， and SLC7A11 proteins in kidney tissue of the mice in various groups. Results The HE staining results showed that compared with control group， there were vacuoles in the renal tubule lumen of the mice in LPS group， the score of renal tubular injury was increased significantly（P<0.01）； compared with LPS group，the renal tubular epithelial cells in kidney tissue of the mice in LPS+rosiglitazone group were closely arranged， there was almost no protein tube， and the score of renal tubular injury of the mice was decreased significantly（P<0.01）.Compared with control group，the levels of CRE and BUN in serum of the mice in LPS group were increased（P<0.01）； compared with LPS group， the levels of CRE and BUN in serum of the mice in LPS+rosiglitazone group were decreased significantly（P<0.01）.The RT-qPCR and Western blotting results showed that compared with control group，the expression levels of ACSL4 mRNA and protein in kidney tissue of the mice in LPS group were increased（P<0.01），the expression levels of GPX4 and SLC7A11 mRNA and proteins were decreased（P<0.01），and the expression levels of IL-6， IL-1β， and TNF-α mRNA were increased（P<0.01）； compared with LPS group， the expression levels of ACSL4 mRNA and protein in kidney tissue of the mice in LPS+rosiglitazone group were decreased（P<0.01），the expression levels of GPX4 and SLC7A11 mRNA and proteins were increased（P<0.01），and the expression levels of IL-6， IL-1β，and TNF-α mRNA were decreased（P<0.01）. Conclusion Rosiglitazone can ameliorate the ferroptosis of renal tubular epithelial cells in the AKI mice induced by LPS through regulating ACSL4.

Objective To investigate the effect of carnosic acid （CA） on the endoplasmic reticulum stress （ERS） in the depression rats induced by chronic unpredictable mild stress （CUMS）， and to elucidate its antidepressant mechanism. Methods Twelve SD rats were randomly selected as control group from 66 SD rats by random number table method， and the remaining 54 rats were induced to establish depression model by CUMS combined with solitary feeding， and 48 rats were randomly divided into model group， fluoxetine（3.17 mg·kg^－1） group， CA （40 mg·kg^－1） group， CA （40 mg·kg^－1）+ EX527 silence information regulator 1 （SIRT1） inhibitor （5 mg·kg^－1） group， and there were 12 rats in each group. After corresponding drug intervention， the total exercise distance and intermediate residence time of the rats were measured by open field test （OFT）； the immobility time of the rats during forced swimming was observed by forced swimming test （FST）； the surge water perference of the rats in various groups were observed by sugar water preference test （SPT）； the percentages of the TUNEL positive cells in hippocampus tissue of the rats in various groups were detected by TUNEL method； immunofluorescence method was used to detect the positive expression intensities of C/EBP-homologous protein （CHOP） and glucose regulated protein 78 （GRP78） in hippocampus tissue of the rats in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of SIRT1， GRP78， CHOP， and Caspase-12 mRNA in hippocampus tissue of the rats in various groups； Western blotting method was used to detect the expression levels of SIRT1， GRP78， CHOP Caspase-12， and cleaved Caspase-12 （cleaved Caspase-12） proteins in CA1 region in hippocampus tissue in brain of the rats in various groups. Results Compared with control group， the total exercise distance， intermediate residence time， sugar water preference in OFT， and expression levels of SIRT1 protein and mRNA in CA1 region in hippocampus tissue in brain of the rats in model group were significantly decreased （P<0.05）， the immobility time during forced swimming， percentage of TUNEL positive cells，expression levels of GRP78， CHOP，Caspase-12， and cleaved Caspase-12 proteins were significantly increased （P<0.05）.Compared with model group， the total exercise distance， intermediate residence time，sugar water preference in OFT，and expression levels of SIRT1 protein and mRNA in hippocampus tissue of the rats in fluoxetine group and CA group were significantly increased （P<0.05）， the immobility time during forced swimming， percentage of TUNEL positive cells， expression levels of GRP78， CHOP，Caspase-12， and cleaved Caspase-12 proteins were significantly decreased （P<0.05）；compared with CA group， the total exercise distance， intermediate residence time，sugar water preference in OFT， and expression levels of SIRT1 protein and mRNA in CA1 region in hippocampus tissue in brain of the rats in CA+EX527 group were significantly decreased （P<0.05）， the immobility time during forced swimming， percentage of TUNEL positive cells， expression levels of GRP78， CHOP，Caspase-12， and cleaved Caspase-12 proteins were significantly increased （P<0.05）. Conclusion CA can alleviate the depression-like behaviors of the rats caused by CUMS， and its antidepressant mechanism may be related to the inhibition of ERS mediated by SIRT1.

Objective To explore the protective effect of velvet antler peptide （VAP） pretreatment on the myocardial H9c2 cell injury of the rats induced by tert-butyl hydroperoxide（TBHP）， and to clarify the effect of VAP on miR-133a/transforming growth factor-β（TGF-β）/Smad axis and its mechanism. Methods The H9c2 cells were divided into blank control group， blank serum group， TBHP group， TBHP+low dose（100 mg·kg^－1） of VAP group， TBHP+ high dose（400 mg·kg^－1） of VAP group， and miR-133 inhibitor（miR-133 inhibitor） group. The cells in blank control group were given no treatment， and the cells in the other groups were treated with VAP for 24 h， then were treated with 200 μmol·L^－1 TBHP； the cells in miR-133 inhibitor group were transfected with miR-133 inhibitor for 24 h，treated with 100 mg·kg^－1 VAP for 24 h+200 μmol·L^－1 TBHP. MTT assay was used to detect the survival rates of the H9c2 cells in various groups；the levels of troponin T（cTnT）， and cardial troponin T（cTnI）in culture supernatant of the cells in various groups were detected by enzyme-linked immunosorbent assay（ELISA） method and creatine kinase-MB（CK-MB）； the expression levels of miR-133 in the H9c2 cells in various groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method；the expression levels of transforming growth factor-β1（TGF-β1），phosphorylated Smad2/3（p-Smad2/3）， and Smad4 proteins the in H9c2 cells in various groups were detected by Western blotting method. Results The MTT results showed that compared with TBHP group， the survival rates of H9c2 cells in TBHP+low dose of VAP group and TBHP+high dose of VAP group were increased （P<0.05）， but the survival rate of the H9c2 cells in miR-133 inhibitor group had no significant difference （P>0.05）. The ELISA assay results showed that compared with blank control group the levels of cTnT， cTnI，and CK-MB in the H9c2 cells in TBHP group were increased （P<0.05）； compared with TBHP group， the levels of cTnT， cTnI，CK-MB in the H9c2 cells in TBHP+low dose of VAP group and TBHP+high dose of VAP group were decreased （P<0.05）.The RT-qPCR results showed that compared with blank control group， the expression level of miR-133a in the H9c2 cells in TBHP group was decreased（P<0.05）；compared with TBHP group， the expression levels of miR-133a in the H9c2 cells in TBHP+ low dose of VAP group and TBHP+ high dose of VAP group were increased （P<0.05 or P<0.01），and the expression level of miR-133a in the cells in miR-133 inhibitor group was decreased（P<0.01）. The Western blotting results showed that compared with blank control group， the expression levels of TGF-β1， p-Smad2/3， and Smad4 proteins in the H9c2 cells in TBHP group were increased（P<0.05）； compared with TBHP group， the expression levels of TGF-β1， p-Smad2/3， and Smad4 proteins in the H9c2 cells in TBHP+low dose of VAP and TBHP+ high dose of VAP groups were decreased （P<0.05）. Conclusion VAP pretreatment can protect the myocardial H9c2 cell injury of the rats induced by TBHP through regulating the TGF-β/Smad signaling pathway with miR-133a.

Objective To discuss the becteriostatic effect of flavonoids in the Carex meyeriana Kunth extract on infection of Staphylococcus aureus （S. aureus） of the rats after skin scalding， and to provide the new ideas，for the research on new-style anti-infection traditional Chinese medicine. Methods The water-alcohol extraction method was used to extract the active ingredients of Carex meyeriana Kunth and the content of flavonoids was determined by UV spectrophotometry， Oxford cup bacteriostatic test was used to detect the bacteriostatic activity of active ingredients of the Carex meyeriana Kunth against bacteria， and its minimal inhibitory concentration（MIC） was determined by tube dilution method. A total of 24 SD rats were used to establish Ⅱ degree scalding models and randomly divided into normal saline control group， penicillin sodium （NaBP） control group and Carex meyeriana Kunth extract CM-70 treatment group， and there were 8 rats in each group. The rats in three groups were inoculated with 20 μL （1.0×10⁸ CFU·mL^－1） S.aureus by multi-point injection in the wound skin；on the next day， the rats in three groups were smeared with sterile normal saline （0.5 g sterile absorbent cotton adsorbed normal saline 0.5 mL·cm^－2）， NaBP （0.5 g sterile absorbent cotton adsorbed NaBP， 5 000 U·kg^－1 body mass） and 10 g·L^－1Carex meyeriana kunth CM-70 extract diluent （0.5 g sterile absorbent cotton adsorbed 0.5 mL·cm^－2Carex meyeriana Kunth extract CM-70 extract diluent）. The wound infection of the rats in various groups were observed； the bacteria number （CFU·g^－1）under scalding wound scab of the rats on the 1st， 2nd， 3rd， 4th， 5th， 6th， and 7th days after scalding was measured by bacteria quantitative test； the serum levels of interleukin-1β （IL-1β）， interleukin-10 （IL-10）， and tumor necrosis factor-α （TNF-α） of the rats in various groups before scalding and on the 1st， 3rd， 7th， and 14th days after scalding were detected by ELISA method. Results The in vitro bacteriostatic activity test results showed that different dilution concentrations of Carex meyeriana Kunth extract CM-70 formed different sizes of inhibition zones against S.aureus， and did not formed the inhibition zones against Escherichia and Candida albicans. The MIC detection results showed that the MIC value of active ingredients of the Carex mgyeriana Kunth extract to S. aureus was 2.5 g·L^－1. The observation results of infection and healing of scalding wounds of the rats in various groups showed that compared with normal saline control group， on the 3rd day after scalding， the exudation of wound serous of the rats in Carex meyeriana Kunth extract CM-70 treatment group was less； on the 7th day after scalding， the wound serous exudation， redness and swelling，and the wound area of the rats were decreased； on the 14th day after scalding， the wounds of the rats were completely dry without serous exudation，the scabs were formed on the surface， and the wounds healed well， which was similar to that in NaBP control group. The number of bacteria under scalding wound scab detection results showed that compared with normal saline control group， the number of bacteria under scalding wound scab of the rats in Carex meyeriana Kunth extract CM-70 treatment group was decreased significantly at the same time （P<0.01）. The ELISA results showed that compared with normal saline control group， the level of serum IL-10 of the rats in Carex meyeriana Kunth extract CM-70 treatment group was increased significantly on the 1st day and 3rd day after scalding（P<0.01）， and it was decreased significantly on the 7th day and 14th day （P<0.05）；the serum levels of IL-1β and TNF-α of the rats in Carex meyeriana Kunth extract CM-70 treatment group were decreased significantly on the 7th day and 14th day（P<0.05）. Conclusion The active ingredients of Carex meyeriana Kunth can significantly inhibit the infection of S. aureus after skin scalding in the rats and exert the anti-inflammatory and bacteriostatic effects by regulating the levels of serum IL-10，IL-1β， and TNF-α.

Objective To investigate the effect of silent information regulator 2（SIRT2）on the aerobic glycolysis， growth and proliferation of the colorectal cancer cells， and to clarify its related mechanism. Methods The HCT116 cells and SW480 cells were cultured， and Western blotting method was used to detect the expression levels of SIRT2 protein in two kinds of cells.The HCT116 cells with high expression of SIRT2 protein were selected for the following RNA interference experiment. Negative control group，SIRT2-siRNA#1 group， SIRT2-siRNA#2 group， and SIRT2-siRNA#3 group were established； the SW480 cells with low expression of SIRT2 protein were selected for the following SIRT2 overexpression experiment.The SIRT2 recombinant plasmids were constructed， and the SW480 cells at logarithmic growth phase were selected； blank group， empty vector plasmid transfection group， and SIRT2 plasmid transfection group were established；the recovery experiment was carried out in the HCT116 cells；control group， SIRT2-siRNA+glucose-6-phosphate dehydrogense（G6PD） plasmid co-transfection group and SIRT2-siRNA#3 group were established.The plasmid and siRNA were transfected into the SW480 cells and HCT116 cells by Lipofectamine 2000， respectively，the conterts of glucose in the culture medium in various groups were measured by glucose oxidase method after silencing SIRT2 and over-expression of SIRT2；the levels of lactic acid in the culture medium in various groups were measured by colorimetry after silencing SIRT2 and over-expression of SIRT2；the clone formation rates of the cells in various groups were determined by clonal formation assay； the proliferation activities of the cells in various groups were determined by CCK-8 assay；the expression levels of G6PD mRNA in the cells after silencing SIRT2 and over-expression of SIRT2 were determined by reverse transcriptionPCR（RT-PCR） method；the expression levels of G6PD protein in the cells in various groups after silencing SIRT2 and over-expression of SIRT2 were detected by Western blotting method；the expression levels of SIRT2 and G6PD proteins in colorectal cancer tissue were determined by immunohistochemistry.After co-transfection of SIRT2-siRNA and G6PD plasmids， the glucose levels，lactic acid levels，survival rates， and proliferation abilities of the cells were detected. Results Compared with negative control group， the level of glucose in the culture medium in SIRT2-siRNA#3 group was increased significantly（P<0.05）， the level of lactic acid was significantly decreased（P<0.05），the survival rate and proliferation ability of the cells were significantly decreased（P<0.05），the expression levels of G6PD mRNA and protein were decreased （P<0.05）；compared with empty vector plasmid transfection group，the level of glucose in the culture medium in SIRT2 plasmid transfection group was decreased significantly（P<0.05）， the level of lactic acid was significantly increased（P<0.05）， the proliferation ability and clone formation rate of the cells were significantly increased（P<0.05）.The expression levels of SIRT2 and G6PD proteins in colorectal cancer tissue were significantly higher than that in paracancerous tissue（P<0.05）. Compared with SIRT2-siRNA#3 group， the level of glucose in the culture medium in SIRT2-siRNA+G6PD plasmid co-transfection group was significantly decreased（P<0.05）， the level of lactic acid in the culture medium was significantly increased（P<0.05），and the proliferation ability and clone formation rate of the cells were significantly increased（P<0.05）. Conclusion SIRT2 promotes the aerobic glycolysis and growth and proliferation of the colorectal cancer cells，and its mechanism may be related to the expression of G6PD regulated by SIRT2.

Objective To discuss the influencing factors for the risk of Alzheimer’s disease （AD） after correction the Mini-Mental State Examination （MMSE） score trajectory， and to clarify the risk factors for of AD in the people with different MMSE scores. Methods The follow-up data from 2005－2016 were collected based on the AD Neuroimaging Program Database. After screening， the follow-up data of 425 people were finally included， and the variables were screened by LASSO method； the influencing factors of MMSE scores at different quantiles were analyzed by Bayesian quantile regression model， and COX model and Bayesian quantile regression joint model were used to analyze the main risk factors of AD. Results After screening， 10 variables were included， including albumin， total cholesterol， and blood glucose concentration and so on. The results of longitudinal sub-model analysis of the Bayesian quantile regression joint model showed that the influencing factors which affecting the change of MMSE score trajectory at different quartiles were the same， such as albumin， blood glucose concentration， age， gender，apolipoprotein E（APOE4） gene， race， and educational score. The Cox regression sub-model anslysis of the joint model results showed that race and APOE4 gene had the effects on the AD at all quartiles， and the risk ratios of APOE4 gene were 2.188（（95%CI：1.775，2.620）， 1.751（95%CI：1.422，2.042）， 1.706（95%CI：1.391，2.102）， and 2.056（95%CI：1.439，3.206）.The total cholesterol level and history had the effects on the AD only at parts of quartiles. Conclusion The risk factors of AD are different in the people with different MMSE score distributions， and the degree of influencing are also different. Both APOE4 gene and whiteness are the risk factors of AD at different quartiles， and the total cholesterol level and history are the risk factors of AD only at parts of quartiles.

Objective To screen the differential prognostis ferroptosis genes of thyroid cancer （TC） and construct the prognostic risk model of TC ferroptosis related genes（FRGs）， and to clarify its potential mechanism at the molecular level. Methods The gene expression and clinical data were obtained from The Cancer Genome Atlas （TCGA） Database. The FRGs were obtained from FerrDb and GeneCards Databases， and R software was used to screen the PFRGs of TC；the PFRGs mRNA expressions in TC and thyroid tissues were obtained from TCGA and GTEx Databases， and the immunohistochemical results were obtainted from the Human Protein Atlas （HPA） Database to verify the differences in the expressions of PFRGs mRNA and protein； time-receiver operating characteristic （time-ROC） curve and Kaplan-Meier curve were used to evaluate the relationships between PFRGs and survival and prognosis of the TC patients； univariate and multivariate Cox regression analysis were used to calculate the risk scores of PFRGs expression， the clinical data of the TC patients were included， the independent prognostic analysis was performed， and the Nomogram chart was constructed. Spearman correlation analysis was used to obtain the correlation between the expressions of PFRGs and the expressions of other genes in TCGA Database and expressions of various genes，the correlation coefficient was calculated and the co-expressing genes were screened；the co-expression genes of PFRGs were analyzed by protein-protein interaction（PPI） network diagram，Geno Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis. Results A total 3 317 up-regulated genes and 3 456 down-regulated genes in TC were screened out by differential analysis；343 differentially expressed genes（DEGs） screened out by univariate Cox regression were associated with the survival and prognosis of the TC patients， including CD44， Annexin A1（ANXA1）， and nuclear receptor subfamily 4 group A member 1（NR4A1）. The Kaplan-Meier and time-ROC curves results showed that the expressions of CD44， ANXA1， and NR4A1 were associated with the survival and prognosis of the TC patients （P=0.048， P=0.005， P=0.036）， and all had good 1-year， 3-year， and 5-year survival prediction effects；the risk scoring system of three genes was constructed to calculate the risk score，and the risk score was a prognostic factor of the TC patients ［hazard ratio（HR）=8.882， 95%CI=1.561－50.547，P=0.014］， and the higher the risk score was，the worse the survival prognosis was［P=0.011，area under curve（AUC）=0.761，0.767，and 0.722）； the Nomogram chart （C-index=0.938） constructed by the risk score combined with the clinical characteristics of the TC patients had a good predictive effect on the survival of the TC patients. The co-expression and enrichment analysis results showed that TC ferroptosis and its co-expressing genes （DUSP1， DUSP5， DUSP6，FOS，IL1RAP， JUN， MET， RASGRF1，TGFA，TGFBR1，andTNFRSF1A）mediated the MAPK signaling pathway and affected the MAP kinase/serine/threonine tyrosine phosphatase activity and MAP kinase phosphatase activity，and regulated the inactivation of the MAPK activity. Conclusion The TC differential PFRGs CD44， ANXA1， and NR4A1 based on bioinformatics analysis are related to the survival and prognosis of the TC patients. The prediction model constructed by three genes has a good predictive ability，and its mechanism may be related to the polygenic network’s regulation of the MAPK signaling pathway.

Objective To analyze the relationship between the expression level of COMM domain-containing protein 7 （COMMD7） and prognosis of the brain low-grade glioma （LGG） patients by bioinformatics method， and to find new potential biomarkers of brain LGG， and to establish the prognostic prediction model，and to provide the basis for the prognostic prediction and individualized treatment of the patients. Methods A total of 523 brain LGG samples and 1 152 normal samples were downloaded from the UCSC Genome Database. Mann-whitney U test was used to analyze the expression difference of COMMD7 between brain LGG samples and normal samples. The Human Protein Atlas （HPA） Database was used to verify the results. DESeq software package of R language was used to screen the differentially expressed genes （DEGs） in the brain LGG samples in low expression of COMMD7 group and high expression of COMMD7 group； pROC software package of R language was used to perform the receiver operating characteristic （ROC） curve， univariate and multivariate Cox regression analysis were used to analyze the relationship between the COMMD7 expression and clinicopathological features of the brain LGG patients and its effects on the 1，3， and 5-year survival rates of the brain LGG patients； ggplot2 software package of R language was used to construct the forestplot； RMS software package of R language and survival package were used to construct the Nomogram and Calibration prediction model；the proportional risk regression model was used to analyze the diagnostic value of COMMD7 in distinguishing the brain LGG samples from normal samples. GEPIA and Oncolnc Databases were used to further verify the relationship between COMMD7 and prognosis of the brain LGG patients. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis were used to analyze the function and pathway enrichment of the DEGs； Gene Set Enrichment Analysis （GSEA） was used to obtain the significantly enriched gene sets of DEG； TISIDB Database was used to analyze the correlation between the COMMD7 expression and immune cell infiltration of the brain LGG patients. Results The expression of COMMD7 in the brain LGG samples was significantly increased，and the HPA detection results showed that COMMD7 was highly expressed in the brain LGG samples. A total of 764 DEGs were identified （|log₂FC|>1， P<0.05）， including 654 up-regulated DEGs and 110 down-regulated DEGs.The predictive model based on the multivariate analysis results suggested that COMMD7 was an independent prognostic factor. The ROC analysis results showed that COMMD7 had a high diagnostic value in distinguishing the cancer tissue from the adjacent tissue ［area under curve （AUC） = 0.835， 95%CI = 0.816－0.853］.The Cox regression results showed that the survival rate of the brain LGG patients in high expression of COMMD7 group was significantly lower than that in low expression of COMMD7 group， and the high expression of COMMD7 was negatively related with poor prognosis of the brain LGG patients（P<0.01）. The analysis results of 514 LGG samples from GEPIA Database and 510 brain LGG samples from Oncolnc Database showed that the survival rate of the brain LGG patients in high expression of COMMD7 group was significantly lower than that in low expression of COMMD7 group （P=0.003， P=0.006）； the GO enrichment analysis results showed that the above DEGs were mainly enriched in the DNA-binding transcription activator activity， RNA polymerase Ⅱ-specificity， enhancer sequence specific DNA binding， and enhancer binding and so on； the KEGG enrichment analysis results showed that DEGs were mainly enriched in the transcriptional misregulation pathway in cancer. The GSEA results showed that DEGs were mainly enriched in the cell cycle in KEGG Database（N=1.950， P=0.012），cell cycle in World Press（WP） Database（N=1.944， P=0.012）， G₂ /M checkpoints （N=2.118，P=0.0012），and G₂/M DNA damage checkpoints （N=1.879， P=0.012）. High expression of COMMD7 was not only associated with stabilization of P53 closely realted to tumor（N=1.793，P=0.012），but also activating P53 downstream pathway（N=1.782，P=0.012），and p53 signaling pathway （N=1.762，P=0.012）， and activating Tp53 regulated transcription of cell cycle genes （N=1.766， P=0.018）.The immune infiltration analysis results showed that the expression of COMMD7 was significantly positively correlated with the numbers of 15 kinds of cells including activated CD8+T lymphocytes，central memory CD8+T lymphocyte and CD56^bright natural killer cells， and so on（P<0.05）， the expression of COMMD7 was significantly positively correlated with 11 kinds of key immune checkpoints including indoleamine 2， 3-dioxygenase 1 （IDO1）， galactin 9 （LGALS9）， and CD244， and so on（P<0.05），and the expression of COMMD7 was significantly negatively correlated with CD274 （PD-L1）， kinase insert domain receptor （KDR）， and T lymphocytes immunoreceptor with Ig and ITIM domains （TIGIT） （P<0.05），and the expression of COMMD7 was positively correlated with 4 kinds of key immune stimulators including CD40， CD276， and CD48，and 5 kinds of immune chemokines including C-C motif chemokine ligand 2 （CCL2），C-C motif chemokine ligand 5 （CCL5）， C-X-C motif chemokine ligand （CXCL9） and so on. Conclusion The high expression of COMMD7 may be a factor of the poor prognosis and a potential biomarker and therapeutic target of the brain LGG patients， and it can promote the occurrence and development of brain LGG by regulating the transcriptional misregulation pathway in cancer， activating p53 signaling pathway and the dysregulation of p53 downstream pathway related genes. COMMD7 may play a key role in the immune checkpoint inhibitor therapy in the brain LGG patients.

Objective To discuss the expression level of macrophage scavenger receptor 1 （MSR1） in pan-cancer tissue，survival and immune characteristics of the patients by bioinformatics analysis， and to elucidate the value of MSR1 as a new biomarker for the tumor diagnosis， prognosis， and immunotherapy. Methods Clinical Bioinformatic Database and Sangerbox Database were used to analyze the expression levels of MSR1 mRNA in normal tissue and tumor tissue，the expression of MSR1 protein was analyzed by The Human Protein Atlas（HPA） Database，and univariate survival analysis and Kaplan-Meier analysis were used to evaluate the prognostic value of MSR1，and the expressions of MSR1 in various types of cells were analyzed with single-cell sequencing results from Tumor Immune Single-cell Hub （TISCH） Database；Tumor Immune Estimation Resource （TIMER2.0），Tumor Immune Syngeneic Mouse （TISMO）， Tumor Immune Dysfunction and Exclusion （TIDE）， and Gene Set Cancer Analysis （GSCA） Databases were used to analyze the correlations between the MSR1 expression level in pan-cancer tissue and immune cell infiltration， immune checkpoint gene expression， and immune treatment response. Results Compared with normal tissue， the expression levels of MSR1 mRNA in 17 kinds of tumor tissues including breast invasive carcinoma （BRCA），colon adenocarcinoma （COAD），esophageal carcinoma （ESCA）， glioblastoma multiforme （GBM）， head and neck squamous cell carcinoma （HNSC），kidney renal clear cell carcinoma （KIRC）， kidney renal papillary cell carcinoma （KIRP），brain low-grade glioma （LGG），liver hepatocellular carcinoma （LIHC），ovarian serous cystadenocarcinoma （OV），pancreatic adenocarcinoma（PAAD），prostate adenocarcinoma （PRAD），skin cutaneous melanoma （SKCM），stomach adenocarcinoma （STAD），testicular germ cell tumor （TGCT），thyroid carcinoma （THCA）， and uterine carcinosarcoma （UCS） were increased（P<0.01）；compared with normal tissue，the expression levels of MSR1 protein in breast cancer， endometrial cancer， liver cancer， ovarian cancer， skin melanoma， testis cancer， pancreatic cancer， and prostate cancer tissues were increased in varying degrees. The high expressions of MSR1 in bladder urothelial carcinoma ［BLCA， hazard ratio（HR）=1.01， P=0.047， 95%CI（1.00，1.03）］， LGG ［HR=1.03， P<0.001， 95%CI（1.02，1.04）］， LIHC ［HR=1.04， P=0.007， 95%CI（1.01，1.07）］， OV ［HR=1.02，P=0.028， 95%CI（1.00，1.03）］， STAD ［HR=1.02， P=0.016， 95%CI（1.00，1.04）］， THCA［HR=1.06，P=0.006， 95%CI（1.02，1.11）］ and uveal melanoma ［UVM， HR=1.18，P=0.044， 95%CI（1.00，1.40）］ were correlated with the poor overall survival（OS） of the patients； the high expression of MSR1 in LGG ［HR=1.03，P<0.001， 95%CI（1.02，1.04）］，the uterine corpus endometrial carcinoma （UCEC］， ［HR=1.05， P=0.038， 95%CI（1.00，1.10）］， and UVM ［HR=1.2，P=0.036， 95%CI（1.01，1.41）］ was correlated with poor disease-specific surival（DSS）.The single-cell sequencing results indicated that MSR1 was mainly expressed in the dendritic cell（DC） and monocyte-macrophage. The MSR1 expression was positive correlated with various kinds of immune cell infiltrations（P<0.05），including CD8+ T lymphocytes， natural killing（NK） cells， regulatory T lymphocytes， and cancer associated fibroblasts（CAF）. There were significant positive correlations between MSR1 and classical immune checkpoint gene and immunotherapy response marker （P<0.05）. Conclusion MSR1 is highly expressed in lots of tumor tissues and is closely associated with the poor prognosis and immune factors of lots of tumor patients. MSR1 may be a diagnostic tumor marker and a predictor of tumor prognosis and immunotherapy efficacy， and has the potential to be a new therapeutic target.

Objective To discuss the expression level of gasdermin E （GSDME） in the epithelial ovarian cancer （EOC） tissue， and to clarify the effect of GSDME on the occurrence and development of EOC. Methods The expressions of GSDME mRNA in normal tissue and EOC tissue was detected by R language based on the Cancer Genome Atlas （TCGA） and Genotype-Tissue Expression （GTEx） Databases；the correlation between the expression level of GSDME and the occurrence of EOC was analyzed. The difference analysis， Gene Ontology （GO） analysis，Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway richment analysis， Gene Set Enrichment analysis （GSEA）， protein-protein interaction （PPI） network， HUB gene interaction network，and immune infiltration analysis were used to discuss the genes and signaling pathways associated with GSDME and their effects on the occurrence and development of EOC. The pathological tissue sections of 40 EOC patients were collected and divided into low-grade group （well-differentiated and moderately-differentiated，n=24） and high-grade group （poorly differentiated，n=16） based on the pathological grades； according to the sensitivity to cisplatin （DDP）， the pathological tissue sections were divided into DDP resistant group and DDP sensitive group. In addition， 5 tissue sections of normal ovary， benign ovarian tumor， and 5 tissue sections of borderline ovarian carcinoma were collected as controls. The expression intensities of GSDME in all the tissue sections were detected by immunohistochemistry （IHC） method. Besides， 10 pairs of fresh ovarian cancer and paracancerous tissues were selected， and the expression levels of GSDME protein in the samples in two groups were detected by Western blotting method. The ovarian cancer Skov3 cells and Skov3/DDP cells were cultured in vitro， and the expression levels of GSDME protein in the cells were detected by Western blotting method and the relationship between the expression level and DDP-resistance of ovarian cancer was analyzed. Results The analysis results of unpaired samples in the TCGA and GTEx Databases showed that compared with normal ovarian tissue， the expression level of GSDME mRNA in EOC tissue was significantly decreased （P<0.05）， and the expression of GSDME mRNA in EOC tissue in high-grade group was higher than that in low-grade group（P<0.05）.The GSDME single gene difference analysis results showed that there were 460 up-regulated and 329 down-regulated genes |（Log₂ fold charge（FC）|>1，P<0.05）.The functional enrichment and immune infiltration analysis results showed that high expression of GSDME could upregulate the activities of humoral immunie response pathways，leukocyte migration pathway and immunoglobulin complex pathway mediated by circulating immunoglobulin and promote the infiltration of immune cells in tumor microenvironment. The IHC staining results showed that GSDME was related to the pathological grade and drug resistance of EOC； the expression intensity of GSDME in EOC tisssue in DDP sensitive group was higher than that in DDP resistant group（P<0.05）.The Western blotting results showed that compared with paracancerous tissue， the expression level of GSDME protein in EOC tissue was significantly decreased （P<0.05）； compared with Skov3 cells， the expression level of GSDME protein in the Skov3/DDP cells was significantly decreased （P<0.05）. Conclusion GSDME is closely related to the malignancy and cisplatin resistance of EOC. The low expression of GSDME is a key factor leading to the decreasing of tumor immune infiltration， EOC deterioration and resistance.GSDME may be a potential molecular target for the treatment of EOC.

Objective To identify the genes related to liver injury induced by hexavalent chromium ［Cr（Ⅵ）］ through the bioinformatics method， and to provide the new direction for further research on the mechanism of liver injury induced by Cr（Ⅵ）. Methods The data sets related to “［Cr （Ⅵ）］ liver” in the Gene Expression Omnibus （GEO） Database were searched， the data set GSE65198 was downloaded； the differentially expressed genes （DEGs） were screened through limma algorithm； Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed； STRING Database was used to analyze the protein-protein interaction （PPI） network； Cytohubba plug-in was used to screen the hub genes； the targeted drug molecules were predicted by Drug Characterization Database in Enrichr website. Results Twenty samples from the GSE65198 data set were selected and divided into two groups. In the first group， there were five liver samples 1 d after single exposure to 20 mg·kg^－1 Cr （Ⅵ） and five control samples； in the second group， there were five liver samples 7 d after single exposure to 20 mg.kg^－1 Cr （Ⅵ） and five control samples；the control samples in each group were regarded as the negative control，and 342 and 61 DEGs were screened （|log₂ FC|>1 and P<0.05）. The GO enrichment analysis results showed that the DEGs were mainly enriched in the biological processes such as the drug response， mitosis， senescence and so on； the KEGG analysis results showed that the main signaling pathways included the ribosome biogenesis， cell cycle regulation，and p53 signaling pathway and so on. The hub gene analysis results in the PPI network showed that the EBNA1 binding protein 2 （EBNA1BP2）， nucleolar protein 58 （NOP58）， guanine nucleotide binding protein like 3 （GNL3）， nucleolar protein 56 （NOP56）， and WD repeat domain 12 （WDR12） in 1 d group after single exposure and cyclin B2 （CCNB2） （the identifier in PPI network was ENSRNOP00000017117）， cyclin A2 （CCNA2） cyclin B1 （CCNB1）， cyclin dependent kinase 1 （CDK1）， and kinesin family member 20B （KIF20B） in 7 d group after single exposure were screened as the hub genes. The targeted drug prediction results showed that roscovetine and other drugs were identified as the targeted drugs for liver injury by induced Cr（Ⅵ）. Conclusion The mechanism of liver injury induced by Cr（Ⅵ） may be related to cell cycle regulation-related genes， and roscovitine may be the potential clinical treatment drug.

Objective To detect the expression of miR-4286 in exosome in peripheral blood of the patients with acute myeloid leukemia（AML）， and to clarify the value of miR-4286 in the diagnosis of AML. Methods The peripheral blood samples from 45 newly diagnosed patients with AML （AML group） and 35 concurrent healthy subjects（control group） were collected.The exosomes from serum of the subjects in two groups were obtained by over speend contrifugation； the morphology of the centrifuged products was observed under transmission electron microscope（TEM）；and the particle diameter was detected by nanoparticle tracink analysis（NTA）； the expressions of surface markers CD63 and CD9 proteins were analyzed by Western blotting method；the expression levels of miR-4286 in exosome of the subjects in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method；the receiver operating characteristic （ROC） curve was used to calculate the area under curve （AUC）； the sensitivity and specificity of miR-4286 in the diagnosis of AML were analyzed. Results The TEM results showed that the extracted exosomes in peripheral blood showed a membrane structure with roundness in apperance and concave in the middle， and the diameter was about 30－100 nm；the NTA results showed that the particle sizes of exosomes were mainly distributed at 30－150 nm；the Western blotting results showed that the exosomes all expressed CD63 and CD9 proteins，with the typical characteristics of the exosomes.The RT-qPCR results showed that the expression level of exosome miR-4286 in peripheral blood of the patients in AML group was significantly higher than that in control group（Z=－5.543，P<0.01）. The ROC curve analysis showed that when the critical value of expression level of miR-4286 in exosome was 3.182， the AUC was 0.862 9（95%CI：0.774 7，0.937 1， P<0.01）， the sensitivity was 66.67%，the specificity was 97.14%，and the diagnostic efficacy was good. Conclusion The miR-4286 in exosome in peripheral blood has high diagnostic efficiency for AML；it can be used as a diagnostic marker for AML， and has high clinical diagnostic value.

Objective To detect the expression level of microRNA-92a-3p （miR-92a-3p） in the exosome in serum exosome of the patients with acute myeloid leukemia （AML）， and to explore its diagnostic value for AML. Methods A total of 51 patients with AML （AML group） and 34 physically healthy volunteers during the same period（control group） were selected as subjects，and the peripheral blood specimens of the subjects were collected，and the peripheral blood specimens were collected from the subjects. The serum exosomes were extracted by differential centrifugation，and identified by transmission electron microscope， nanoparticle analyzer and Western blotting method；the expression levels of miR-92a-3p in serum exosome of the subjects in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method；according to the expression level of miR-92a-3p in serum exosome， the receiver operating characteristic（ROC） curve was drawn and the area under the curve （AUC） was calculated to determine the critical value， and the diagnostic efficiency of expression level of miR-92a-3p to AML was evaluated based on the sensitivity and specificity. Results The extracted exosomes had membrane structures with round appearance and concave in the middle under transmission electron microscope； the size of the exosomes was 30－100 nm detected by nanoparticle analyzer；the expressions of CD63 and CD9 proteins were found by Western blotting method.Compared with control group，the expression level of miR-92a-3p in serum exosome of the patients in AML group was significantly decreased（P<0.01）；the ROC curve results showed that the optimal critical value of serum exosome miR-92a-3p for the diagnosis of AML was 0.51， the sensitivity was 76.47%， the specificity was 94.12%，the AUC was 0.913，95% confidence interval（CI）=（0.856，0.971）（P<0.001）. Conclusion The expression level of miR-92a-3p in serum exosome of the AML patients has high sensitivity and specificity in the diagnosis of AML， and it can be used in the clinical diagnosis of early AML.

Objective To discuss the expression of apoptosis-related gene CD44 in the thyroid cancer （THCA） tissue and its relationships with the clinicopathological characteristics of the patients and tumor infiltrating leukocytes（TILs）， and to provide new research directions for the diagnosis and treatment of THCA. Methods The expression spectrum of differentially expressed genes （DEGs） related to THCA apoptosis were obtained from The Cancer Genome Atlas （TCGA） Database. The up-regulated CD44 gene was selected and the expression level of CD44 mRNA in THCA tissue and parancancerous tissue were detected； receiver operating characteristic （ROC） curve was used to identify the THCA tissue and paracancerous tissue； Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and Gene Set Enrichment Analysis （GSEA） were used to analyze the function and pathway enrichment of DEGs； the protein-protein interaction （PPI） network was established by using STRING Database and visualized by Cytoscape software； the relationship between the expression of CD44 mRNA in THCA tissue and the infiltration abundance of TILs was analyzed by TIMER Database； the relationships between expression of CD44 protein and immune checkpoints were analyzed by R software. A total of 110 samples of THCA patients were selected， the expressions of CD44 protein in THCA tissue and paracancerous tissue were detected by the immunohistochemical SABC method. Results The TCGA Database analysis results showed that compared with paracancerous tissue，the expression level of CD44 mRNA in THCA tissue was increased（P<0.05）； the ROC curve analysis results showed that when the critical value was 6.855， the sensitivity， specificity， and accuracy of CD44 were 89.71%，75.13%， and 72.91%， respectively. The GO and KEGG analysis results showed that the DEGs were mainly enriched in the apoptosis-related pathways； the GSEA analysis results showed that the DEGs were associated with degranulation response of the neutrophils.The TIMER Database analysis results showed that the expression level of CD44 mRNA in THCA tissue was positively correlated with the infiltration abundance of B cells， macrophages， CD4+T lymphocytes， dendritic cells， and neutrophils （partial.cor>0，P<0.01），and was negatively correlated with tumor purity and the infiltration adundance of CD8+ T lymphocytes（partial.cor≤0，P<0.01）；the expression of CD44 protein in THCA tissue was positively correlated with 9 immune checkpoint genes （P<0.01）， and negatively correlated with 1 immune checkpoint gene （P<0.01）； the immunohistochemical SABC results showed that the positive expression rate of CD44 protein in THCA tissue was higher than that in paracancerous tissue （P<0.01）； the expression of CD44 protein in THCA tissue was correlated with the tumor diameter and lymph node metastasis （P<0.05），and was not correlated with gender， age， and extra glandular invasion of the patients（P>0.05）. Conclusion High expression of CD44 in THCA tissue may be related to progression and multiple immune responses of tumor.

Objective To investigate the expression of a disintegrin and metalloproteinase 10 （ADAM10） in vascular tissue at the stenosis of human arteriovenous fistula （AVF） and its effect on the proliferation and migration of the vascular smooth muscle cells（VSMCs）， and to clarify its possible molecular mechanism. Methods The stenotic venous vascular tissue of 42 patients with end-stage renal disease （ESRD） who underwent reoperation due to AVF stenosis（AVF group） and their normal venous vascular tissue during the first operation （normal control group） were collected.Immunohistochemistry assay was used to detect the expressions of ADAM10 protein in venous vascular tissue of the patients in two groups；real-time fluorescence quantitative PCR（RT-qPCR） assay was used to detect the expression levels of ADAM10 mRNA in venous vascular tissue of the patients in two groups；the VSMCs were divided into control group and model group ［lipopoly saccharide（LPS） group，given 10 mg·L^－1 LPS］， LPS + si-NC group （transfected with si-NC plasmid +10 mg·L^－1LPS）， LPS+si-ADAM10 group （transfected with si-ADAM10 plasmid +10 mg·L^－1 LPS），LPS+Notch signal pathway inhibitor DAPT group （10 mg·L^－1 LPS+10 μmol·L^－1 Notch signal pathway inhibitor DAPT）， and LPS+si-ADAM10+DAPT group （transfected with si-ADAM10 plasmid +10 mg·L^－1 LPS+10 μmol·L^－1 DATP）； CCK-8 method was used to detect the proliferation activities of the cells in various groups； Transwell assay was used to detect the numbers of the migration cells in various groups； Western blotting method was used to detect the expression levels of ADAM10，proliferating cell nuclear antigen （PCNA），matrix metalloproteinase-9 （MMP-9）， Notch homolog protein 1 （Notch1）， Notch intracellular domain （NICD）， and hairy and enhancer of split 1 （Hes1） proteins in the cells in various groups. Results Compared with normal control group， the expression levels of ADAM10 protein and ADAM10 mRNA in venous vascular tissue of the patients in AVF group were significantly increased （P<0.05）. Compared with control group， the proliferation activity，number of the migration cells and expression levels of ADAM10， PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS group were significantly increased （P<0.05）； compared with LPS and LPS+si-NC group， the proliferation activity and number of the migration cells and expression levels of ADAM10， PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10 group were significantly decreased （P<0.05）. Compared with LPS group， the proliferative activities，numbers of the migration cells and expression levels of PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10 group and LPS+DAPI group were significantly decreased （P<0.05）； compared with LPS+si-ADAM10 group， the proliferation activity， number of the migration cells and expression levels of PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10+DAPT group were significantly decreased （P<0.05）. Conclusion ADAM10 is highly expressed in the vascular tissue at the stenosis of AVF，and silencing the expression of ADAM10 gene can inhibit the proliferation and migration of the VSMCs induced by LPS；its mechanism may be related to blocking the Notch signal pathway.

Objective To detect the levels and existing forms of vitamin D （VD） in peripheral blood of the children with development language disorder（DLD） comorbid with attention deficit hyperactivity disorder（ADHD），to clarify the role of VD in the occurrence of DLD comorbid with ADHD， and to provide the new method for its treatment. Methods A total of 90 children with DLD comorbid with ADHD were regarded as observation group， and fifty physically and mentally healthy children in the Health Examination Center in our hospital during the same period were selected as control group. The DLD and ADHD were diagnosed according to the relevant chapters of the International Classification of Diseases， 11th Revision （ICD-11） and the American Diagnostic and Statistical Manual of Mental Disorders， Fifth Edition （DSM-5）.Griffiths Mental Development Scales （GMDS） was used to detect the language ability and the Developmental Quotient（DQ） was caculated；ADHD was classified according to the Snoopy Rating Scale Ⅳ （SNAP-4） Parent and Teacher Questionnaire；Conners Parent Symptom Questionnaire （PSQ） was used for ADHD scoring；the levels of serum VD of the subjects in two groups were detected by liquid chromatography-mass spectrometry. The results of GMDS and PSQ of all subjects and SNAP-4 of the subjects in observation group were collected.When DQ was less than （100－1）×standard deviation， the language ability of the subjects was lower than that of the contemporary；the children’s behavioral problems were divided into six factors according to the PSQ，such as learning problem， psychosomatic problem， anxiety， hyperactivity-impulsivity， conduct problem， and hyperactivity index； when the scores of six factors were all greater than 2 points， the related problems could be diagnosed；according to SNAP-4， ADHD was divided into predominantly inattentive ADHD（ADHD-I），predominantly-hyperactivity/impulsivity（ADHD-HI） and combined ADHD（ADHD-C） groups. Results In observation group， 71.1% were male and 28.9% were female. The levels of serum VD3 and VD of the chinldren in observation group were significantly lower than those in control group （P<0.01）.The levels of serum VD and VD3 of all three types（DLD-ADHD-C，DLD-ADHD-HI，and DLD-ADHD-I） of the subjects in observation group were significantly lower than those in control group （P<0.01）.The DQ values of the subjects in control group were positively correlated with the levels of serum VD and VD3（r=0.512，P<0.01；r=0.529，P<0.01）；the DQ values of the subjects in observation group was positively correlated with the levels of serum VD and VD3（r=0.299， P<0.01； r=0.279， P<0.01），and was negatively correlated with the level of VD2（r=－0.122， P<0.01）.There were significantly differences in the scores of learning problem， psychosomatic problem， conduct problem，anxiety， hyperactivity/impulsivity and hyperactivity index among three types （DLD-ADHD-C，DLD-ADHD-HI，and DLD-ADHD-I） of the subjects in observation group（P<0.01）.The level of serum VD3 was negatively correlated with the scores of the above indexes of the chlildren with DLD-ADHD-C type in observation group（r=－0.438，r=－0.357，r=－0.422，r=－0.465，r=－0.583， r=－0.593，P<0.01），and was positively correlated with the conduct problem of the children with DLD-ADHD-HI type in observation group（r=0.522，P<0.01），the level of VD3 was negatively correlated with hyperactivity-impulsivity and hyperactivity index（r=－0.455，P<0.05； r=－0.424，P<0.01），was negatively correlated psychosomatic problems and hyperactivity-impulsivity（r=－0.468，r=－0.496，P<0.05）， and was positively correlated with hyperactivity index（r=0.694，P<0.01）. The level of VD was negatively correlated with the scores of all six factors mentioned above of the children with DLD-ADHD-C type in observation group（r=－0.444，r=－0.498，r=－0.450，r=－0.501，r=－0.594， r=－0.522，P<0.01），and was negatively correlated with psychosomatic problem， conduct problem，hyperactivity/impulsivity and hyperactivity index of the children with DLD-ADHD-HI type in observation group（r=－0.355，r=－0.578，r=－0.509， r=－0.422，P<0.05or P<0.01）；the level of VD was negatively correlated with psychosomatic problem and hyperactivity/impulsivity（r=－0.485，r=－0.497，P<0.05）， and was positively correlated with the hyperactivity index （r=0.682，P<0.01）. Conclusion VD3 deficiency may be one of the causes of DLD comorbid with ADHD.VD3 supplementation may have a positive effect in the prevention and treatment of the children with DLD comorbid with ADHD.

Objective To discuss the relationship between the different serum levels of anti-mullerian hormone （AMH） and ovarian response and in vitro fertilization-embryo transfer （IVF-ET）-assisted pregnancy outcome of the infertile patients with polycystic ovary syndrome （PCOS）， and to clarify the influence of serum AMH level on assisted pregnancy outcome of the PCOS patients， and to provide the basis for the selection of assisted pregnancy treatment in the PCOS patients. Methods A total of 1 837 patients aged 21－39 years old who underwent IVF-ET-assisted pregnancy were selected and divided into PCOS group （n=910） and non-PCOS group （n=927） according to whether they were diagnosed as PCOS； a total of 437 patients in PCOS group received fresh single high-quality blastocyst transfer， and they were divided into AMH< 7 μg·L^－1 group and AMH≥7 μg·L^－1 group according to their serum AMH levels；the patients were divided into 20－24 years old group， 25－29 years old group， 30－34 years old and 35－39 years old group according to their ages.Age，serum levels of AMH， gonadotropin（Gn） dosages， estradiol （E2） values on the human chorionic gonadotropin （HCG） day， number of retrieved oocytes，cancellation transfer rates for preventing ovarian hyperstimulation syndrome （OHSS） and clinical pregnancy rates of the patients in PCOS group and non-PCOS group were compared；the Gn dosages， E2 values on the HCG day， number of retrieved oocytes，cancelation transfer rates of preventing OHSS and clinical pregnancy rates of the patients with different AMH levels in PCOS group were analyzed；the serum AMH levels of the patients with different ages in pregnant group and non-pregnant group were compared. Results The level of serum AMH and E2 _{value on the HCG day， number of retrieved oocytes and cancelation transfer rate for preventing OHSS of the patients in PCOS group were higher than those in non-PCOS group （P<0.05）， while the Gn dosage and clinical pregnancy rate were lower than those in non-PCOS group （P<0.05）.The serum AMH level of the patients in PCOS group was positively correlated with the E2 value on the HCG day and number of retrieved oocytes（r=0.502，P=0.001；r=0.233，P=0.001）， and negatively correlated with Gn dosage（r=－0.400，P=0.001）.In PCOS group，compared with AMH< 7 μg·L^－1 group， the E2 value on the HCG day， number of retrieved oocytes， and cancelation transfer rate for preventing OHSS of the patients in PCOS group were increased （Z=－3.952， P=0.001； Z=－2.858， P=0.004； χ² = 23.154， P=0.001）； the Gn dosage and clinical pregnancy rate were decreased（Z=－6.727， P=0.001； χ² = 6.640， P=0.010）. The serum AMH level of the patients in pregnant group in PCOS group was lower than that in non-pregnant group（Z=－4.192， P=0.001）； in 25－29 years old group and 30－34 years old group， the serum AMH levels of the patients in pregnant group were lower than those in non-pregnant group（Z=－2.197， P=0.028，Z=－3.700，P<0.001）. Conclusion For the PCOS patients with serum AMH level≥7 μg·L^－1，the pretreatment should be performed to reduce the AMH level before assisted pregnancy. In the process of IVF-ET-assisted pregnancy， the Gn dosage should be well controlled to reduce the E2value on the HCG day， reduce the cancellation transfer rate for preventing OHSS， try to have the chance of fresh embryo transplantation and increase the clinical pregnancy rate， and improve the outcome of assisted pregnancy.}

Objective To analyze the clinical manifestation， imaging feature， treatment option and postoperative standardized management of the patient with retroperitoneal giant lymphangioma， and to improve the clinicians’ understanding of this disease. Methods The clinical data of one patient with retroperitoneal giant lymphangioma were collected.The patients was diagnosed according to the patient’s specialist examination and imaging characteristics， and the selection of treatment measures and postoperative standardized management were analyzed. Results The patient， a 33-year-old woman， was admitted to the hospital due to “abdominal distension for 2 months”. The specialist examination results showed the abdomen was slightly swelling， without other obvious abnormalities； the full abdominal percussion was mainly voiced， and the abdominal percussion changed into dulled drums at the midaxillary line，the mobile dullness was negative， the bowel sound was normal， and there were no water sound. There was a lump with the size of 17 cm×9 cm×26 cm which was soft， unclear boundary， inactive， feminine， negative for tenderness， the auscultation did not semell and vascular murmurs. The gynecological color ultrasound results showed that a large anedomic area was detected above the uterus reaching up to the lower part of the process，with a width of 18.6 cm， irregular morphology and with separations inside. The computed tomography（CT） results of the abdomen showed that the left renal pelvis and the upper segment of ureter expended，and the watery density opacity could be seen inside. The massive cystic mass can be seen in the abdominal cavity， about 17.9 cm×9.0 cm×26.7 cm， the cyst nodules shadow could be seen locally， with slight enhancement on the enhanced scan， the punctate calcification could be seen locally in the focus， and the other abnormalities were not found. The intraperitoneal cystic and space occupying in the abdominal cavity was considered， excluding the compression of the left ureter， secondary expansion and hydronephrosis of the left kidney and left upper segment of ureter.The abdminal neoplasm resection was performed， and the patient was diagnosed as retroperitoneal giant lymphangiome the surgical process was still smooth， and the postoperative standardized management was performed and the patient was discharged from the hospital. Conclusion The clinical manifestations of retroperitoneal giant lymphangioma are poor. At present， the most valuable imaging diagonsis method of retroperitoneal giant lymphangioma is CT and magnetic resonance imaging （MRI）， and surgical treatment is the preferred treatment measure.

Objective To understand the psychological stress situation of the nursing staffs in a Class A Tertiary Hospital in Changchun City in Jilin Province and analyze its causes， and to provide the scientific basis for improving the psychological stress situation of the nursing staffs and promoting the quality of medical services. Methods The nursing staffs who worked in a Class-A Tertiary Hospital in Changchun City in Jilin province were regarded as the subjects， and the data were collected by Wenjuanxing Network Questionnaire，the Kessler（K10） Scale was used to evaluate the psychological stress levels of the nursing staffs，the Stressor Scale was used to analyze the main stressors， and multivariate Logistic regression analysis was used to analyze the influencing factors of psychological stress levels of the nursing staffs. Results A total of 691 valid questionnaires were collected； the mean K10 score of the nursing staffs was （23.31±8.16） points，and 56.87% of the nursing staffs were under high psychological stress level （K10 score ≥22 points）； there was significant difference in the detection rate of high psychological stress level among the nursing staffs with different ages（P<0.05）. The multivariate Logistic regression analysis results showed that the main risk factors of psychological stress level inccreasing of the nursing staffs were night shift （ OR=1.930， 95% CI： 1.299－2.869，P=0.001） and being attacked or verbally abused by the patients and their family members （OR=3.945， 95% CI： 2.829－5.502，P<0.001）； the total score and the scores on various dimensions of Stressor Scale of the nursing staffs with the high levels of psychological stress were higher than those with the low or average levels of psychological stress（P<0.05）. Conclusion The levels of psychological stress of the nursing staffs in our hospital and the wide range of stressors is wild； the corresponding measures should be put forward according to the workload and doctor-patient contradictions to reduce the psychological stress of the nursing staffs and increase their psychological health levels.