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Objective To explore the therapeutic effect and mechanism of bone marrow mesenchymal stem cells(BMSCs) on ischemic stroke in rats. Methods BMSCs were isolated by whole bone marrow adherence method and identified by flow cytometry.The rat model of middle cerebral artery occlusion(MCAO) was established and randomly divided into sham operation group, MCAO group and BMSC group.The rats in each group were scored by mNSS and stained with TTC.Immunohistochemistry, immunofluorescence and Western blot were used to detect the positive expression levels of Caspase-3,MAP-2,GAP-43 and GFAP in the infarct area and surrounding areas of the brain tissues of each group. Results Flow cytometry detected that BMSCs expressed low CD45 and high expression of CD90 and CD29; The mNSS score and the percentage of infarct volume of rats on the 3 rd, 7 th and 14 th day after transplantation of BMSCs all decreased; The Caspase-3 in the infarct area decreased, and the expression of MAP-2,GFAP and GAP-43 increased. Conclusion BMSCs transplantation can promote the recovery of neurological function and reduce the volume of cerebral infarction in MCAO model rats, possibly through mechanisms such as nerve regeneration cell proliferation, inhibition of cell apoptosis, and axon regeneration.

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Objective To explore the role of rat Sesn2/AMPK signal-mediated liver autophagy in the abnormal glucose metabolism caused by intermittent hypoxia and reoxygenation. Methods 48 rats were randomly divided into a control group(NC) and chronic intermittent hypoxia(CIH) group, with 24 rats in each group.The CIH group was placed in an intermittent hypoxia chamber for 8 h/d for 8 weeks, and then fed under normoxia for 4 weeks; The NC group was always fed under normoxia.The levels of fasting blood glucose(FBG),fasting insulin(FINS),liver Sesn2,AMPK,p-AMPK protein, mTOR and LC3 mRNA levels were measured at baseline, 8 weeks of intermittent hypoxia, and 4 weeks of reoxygenation in the two groups of rats. Results At baseline, there was no statistically significant difference in the above indicators between the two groups(P>0.05); at 8 weeks of intermittent hypoxia, compared with the NC group, the levels of FBG,FINS,HOMA-IR and mTOR in the CIH group increased, the levels of Sesn2,p-AMPK/AMPK and LC3 decreased, and the difference was statistically significant(P<0.05); after 4 weeks of reoxygenation, there was no statistically significant difference in the two groups′ indexes(P>0.05). Conclusion Chronic intermittent hypoxia can cause the increase of FBG and FINS in rats, which may be caused by inhibiting Sesn2/AMPK-mediated liver autophagy and causing insulin resistance.

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Objective To investigate the functional role of Bcl-2-associated Athanogenes(BAGs) family in multiple tumors based on TCGA database. Methods 11 057 tumor samples were downloaded from TCGA,and multidimensional analysis of BAGs family(BAG1-BAG6) was performed. Results There were differences in the expression of Bags family in tumor and paracancerous tissues, and the expression of different BAG genes had a statistically significant effect on the survival prognosis of different cancers. Conclusion Bcl-2-associated Athanogenes may be potential diagnostic and prognostic markers for a variety of cancers, and may provide theoretical support for individualized treatment.

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Objective To investigate the effect of the expression of FASTKD4 in osteosarcoma and its effect on proliferation and apoptosis of osteosarcoma cells. Methods The levels of FASTKD4 mRNA were detected by qPCR,and protein was detected by immunohistochemistry in osteosarcoma and paracancerous tissues.The levels of FASTKD4 RNA were detected by qPCR in different osteosarcoma cells, and the osteosarcoma cell Saos-2 was used as the research object.Saos-2 cells were transfected with FASTKD4 small interfering RNA(shFASTKD4 group) and negative control(shCtrl group).qPCR and Western blot were used to detect the FASTKD4 mRNA and protein in transfected Saos-2 cells.The proliferation of transfected Saos-2 cells were detected by multiparametric high-content screening and CCK-8.The colony formation ability and apoptosis of transfected Saos-2 cells were detected by clone formation assay and flow cytometry. Reuslts The results of qPCR and immunohistochemistry showed the expression of the FASTKD4 in osteosarcoma tissues was higher than that in paracancerous tissues(P<0.05).The proliferation and colony formation ability of shFASTKD4 group were suppressed when compared with shCtrl group(P<0.05).The percentage of apoptosis in shFASTKD4 group was higher than that in shCtrl group(P<0.05). Conculsion This study confirmed that FASTKD4 was highly expressed in osteosarcoma tissues, and silencing of FASTKD4 could inhibit proliferation, colony formation of osteosarcoma cell and induce the apoptosis of osteosarcoma cell, We also found that FASTKD4 Knockdown inactivated PI3 K/AKT Signaling pathway in osteosarcoma cells.

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Objective To explore the mechanism of resveratrol in promoting autophagy of endometrial cancer Ishikawa cells. Methods Ishikawa cells were divided into three groups: control, resveratrol and resveratrol+PI3 Kagonist(740 Y-P).The expression levels of LC3,Beclin1,PI3 K and AKT were detected by qRT-PCR and Western Blot method, the expression intensity of LC3 was detected by cell immunofluorescence method, and the autophagy of cells was observed by electron microscope. Results Resveratrol could increase the mRNA and protein levels of Beclin1 and LC3 in cells; the red fluorescence intensity of cytoplasmic LC3 was enhanced; the number of autophagosomes increased.The P-PI3 K and P-AKT protein levels of Ishikawa cells in the resveratrol group were lower than those in the control group, while the autophagy level of the resveratrol+740 Y-P group was lower than that in the resveratrol group. Conclusion Resveratrol may induce autophagy in endometrial cancer Ishikawa cells by inhibiting PI3 K/AKT pathway, thereby inhibiting cell proliferation.Resveratrol may become a new drug for adjuvant treatment of endometrial cancer.

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Objective To analyze the expression of long non-coding RNA TALNEC2 in the middle cerebral artery occlusion(MCAO) model and oxygen-glucose deprivation(OGD) model, and to explore its regulatory mechanism in the process of acute cerebral infarction. Methods Anin vivomouse model of MCAO and anin vitrocell model of OGD were established to induce cerebral ischemic stroke condition.The expressions of TALNEC2,miR-19 a-3 p and c-jun-N-terminal kinase(JNK) were measured by qRT-PCR analysis.The neurological injuryin vivowas investigated by neurological score and TTC staining.Cell apoptosis and inflammatory injury were analyzed by Western blot, flow cytometry and ELISA,respectively.The interaction between miR-19 a-3 p and TALNEC2 or JNK was explored by luciferase activity assays. Results Up regulation of TALNEC2 was observed in MCAO and OGD models.Knockdown of TALNEC2 attenuated the cerebral infarct, neurological injury, apoptosis and inflammatory injury in MCAO mice.Luciferase activity analysis showed that miR-19 a-3 p combined with TALNEC2.Overexpression of miR-19 a-3 p inhibited the apoptosis and inflammatory response in OGD-treated brain microvascular endothelial cell(BMEC),which was attenuated by TALNEC2.JNK was a target of miR-19 a-3 p and its restoration reversed the miR-19 a-3 p-mediated suppression of apoptosis and inflammation.Moreover, TALNEC2 promoted JNK expression by competitively sponging miR-19 a-3 p. Conclusion The knockdown of TALNEC2 attenuates MCAO-or OGD-induced acute cerebral ischemia injury by regulating miR-19 a-3 p/JNK,which provides a new potential strategy for the treatment of acute cerebral infarction.

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Objective To explore the role of miR-193 a-5 p/HOXA7 axis in the process of acute myeloid leukemia(AML) cell proliferation and apoptosis. Methods The bone marrow samples from newly treated AML patients and normal volunteers were collected to detect changes in the expression levels of miR-193 a-5 p and HOXA7 in the bone marrow samples, and the regulatory relationship between miR-193 a-5 p and HOXA7 was analyzed.The AML cell line THP-1 was cultured in vitro and transfected with empty vector, miR-193 a-5 p overexpression lentiviral vector, HOXA7 shRNA lentiviral vector, and simultaneously transfected with miR-193 a-5 p and HOXA7 overexpression lentiviral vector.The proliferation and apoptosis of THP-1 cells and the expression changes of miR-193 a-5 p and HOXA7 in the cells were detected under different intervention methods. Results The expression level of miR-193 a-5 p in bone marrow cells of AML patients was lower than that of normal volunteers, while the expression level of HOXA7 was higher than that of normal volunteers(P<0.01).The analysis of luciferase activity showed that miR-193 a-5 p could negatively regulate HOXA7.The expression of HOXA7 in THP-1 cells transfected with miR-193 a-5 p overexpression lentiviral vector was lower than that in empty vector cells(P<0.01).Compared with cells transfected with empty vector, transfection of miR-193 a-5 p overexpression lentiviral vector or transfection of HOXA7 shRNA lentiviral vector inhibited the proliferation of THP-1 cells and promoted cell apoptosis(P<0.05).The THP-1 cells transfected with miR-193 a-5 p and HOXA7 overexpressing lentiviral expression vector at the same time proliferated stronger than empty vector cells(P<0.05). Conclusion The expression of miR-193 a-5 p in bone marrow cells of AML patients is significantly reduced, which can promote THP-1 cell apoptosis by inhibiting the expression of HOXA7 and the proliferation of THP-1 cells.

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Objective To explore the difference of gene expression between clear cell renal cell carcinoma and adjacent tissues, and to explore the role of differentially expressed genes in the occurrence and development of clear cell renal carcinoma. Methods High-throughput sequencing was used to analyze clear cell renal cell carcinoma and adjacent tissues.The genes related to metabolism were verified by real-time fluorescence quantitative PCR and Western blot. Results Bioinformatics showed that the expression of 40 ATP synthase-related genes in clear cell renal carcinoma increased.Real-time PCR and Western blot verified this result.Compared with adjacent tissues, the relative abundance of ATP5 F1 mRNA and protein expression of ccRCC increased(P<0.05). Conclusion Clear cell renal carcinoma may lead to changes in the expression of a variety of genes, among which the increased expression of ATP synthase may play an important role in the occurrence and development of clear cell renal cell carcinoma.

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Objective To study the effect and mechanism of transcription factor SPIBon proliferation, migration and invasion of esophageal cancer cells after regulating the expression of SPIB protein by gene expression regulation technology. Methods The expression levels of SPIB mRNA and protein in esophageal cancer cell lines were detected by Western blot and qRT-PCR, respectively. SPIB expression was down-regulated by siRNA in KYSE150 cells, and the proliferation and cell cycle progression were detected by Cell Counting Kit-8 and flow cytometry, respectively. The KYSE30 cell lines with stable overexpression of SPIB were established and the effects of SPIB on cell proliferation and clone formation were detected. The effects of SPIB on the migration and invasion of esophageal cancer cells were detected by cell scratch and Transwell system, and the expression levels of N-cadherin and E-cadherin proteins were detected by Western blot. Results SPIB was highly expressed in esophageal cancer cell lines. Down-regulation of SPIB could inhibit cell proliferation and cell cycle progression, and decrease the expression of Cyclin D1 proteins. Overexpression of SPIB enhanced the proliferation and clone formation of esophageal cancer cells. Knockdown of SPIB protein expression weakened the migration and invasion ability of esophageal cancer cells, and reduced the expression of N-cadherin protein. Conclusion SPIB, as a pro-cancer factor, can promote the proliferation, migration and invasion of esophageal cancer cells.

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Objective To explore the protective effect and mechanism of dapagliflozin on palmitic acid-induced human renal tubular epithelial cells. Methods Human renal tubular epithelial cells were culturedin vitroand divided into normal control(CON) group, palmitic acid(PA) group, palmitic acid+dapagliflozin(PA+Dapa) group, and dapagliflozin(Dapa) group.After 24 hours of intervention, CCK-8 method was used to detect cell proliferation, DCF immunofluorescence was used to detect intracellular reactive oxygen species(ROS),JC-1 fluorescence was used to detect mitochondrial membrane potential, the kit detected the levels of superoxide dismutase(SOD) and malonaldehyde(MDA) in cells, Western blot detected the levels of apoptotic proteins caspase3,Cleaved caspase3 and cytochrome C(Cyto C),and Hoechst33258 fluorescence detection apoptosis. Results Compared with CON,the PA group decreased cell proliferation, increased ROS level, decreased mitochondrial membrane potential, decreased SOD level and increased MDA level, increased levels of apoptotic proteins caspase3 and Cleaved caspase3,and increased expression of Cyto C,the apoptosis level detected by Hoechst33258 increased, and the differences were statistically significant(P<0.05);the above indicators in the Dapa group were not significantly different from those in the CON group; compared with the PA group, the cell proliferation in the PA+Dapa group increased.The level of ROS decreased, the level of mitochondrial membrane potential increased, the level of SOD increased, the level of MDA decreased, the expression of apoptotic proteins caspase3,Cleaved caspase3,and Cyto C decreased, and the level of apoptosis detected by Hoechst33258 decreased.The difference was statistically significant(P<0.05). Conclusion Dapagliflozin has a protective effect on human renal tubular epithelial cells induced by palmitic acid.The mechanism may be related to the improvement of mitochondrial function and the level of oxidative stress.

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Objective To investigate the influence of C-C motif chemokine ligand 20(CCL20) on migration, invasion and proliferation of esophageal squamous cell carcinoma(ESCC) cells, and to preliminarily research its molecular mechanism. Methods The CCL20 expression plasmid and interference plasmid was transfected into ESCC cells, and the stably transfected cells was selected with G418,Western blot or qRT-PCR assay was used to assess the effects of over-expression and interference.The influence of CCL20 in migration, invasion and proliferation of ESCC cells was detected by Transwell and CCK8 assays, pathway inhibitors was used to seek the downstream signaling pathway of CCL20. Results CCL20 overexpressing and interfering cell lines was successfully established, CCL20 overexpression promoted ESCC cells migration and invasion, and increased the activation level of ERK1/2.Interfering CCL20 expression reduced the ability of migration and invasion in ESCC cells, and the activation level of ERK1/2 decreased.Moreover, ERK1/2 specific inhibitor treatment reversed the migration and invasion induced by CCL20. Conclusion CCL20 promotes ESCC cells migration and invasion by activating ERK pathway.CCL20 may be a potential target for the treatment of ESCC.

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Objective This study was to explore the protective effect and mechanism of caffeic acid on adriamycin-induced cardiomyocyte damage. Methods Adriamycin(ADM) is used to induce H9 c2 injury model of cardiomyocytes and to determine the best concentration of it. Then the injured H9 c2 were co-cultured with different concentrations of caffeic acid(CA) for 24 hours, and the optimal concentration of caffeic acid was selected. The H9 c2 were divided into model group(ADM), treatment group(ADM+CA) and blank control group(control). As shown, the survival rate of cardiomyocytes was measured by MTT method, and the morphology of H9 c2 was observed by HE. Then the levels of lactate dehydrogenase(LDH) and reactive oxygen species(ROS), changes in mitochondrial membrane potential and the activities of Na+-K+-ATPase and Ca2+-ATPase in H9 c2 were measured, as well as the expression of NF-κB-p65 were detected by Western blot. Results 2.0 μmol/L ADM and 4.0 μmol/L caffeic acid were selected for subsequent experiments. Compared with the control group, in the adriamycin-treated model group, the cardiomyocyte fiber arrangement was disordered, and the muscle fiber was deformed seriously; LDH and ROS levels were increased, mitochondrial membrane potential were increased, and Na+-K+-ATPase and Ca2+-ATPase activities were reduced. Compared with the model group, in the caffeic acid treatment group, myocardial cell muscle bundles were arranged neatly, LDH and ROS levels, and mitochondrial membrane potential were reduced(P<0. 05), while mitochondrial Na+-K+-ATP and Ca2+-ATPase activities were increased(P<0. 05), NF-κB-p65 protein expression was reduced(P<0. 05).Conclusion Caffeic acid can protect against oxidative stress, reduce mitochondrial membrane potential, increase the level of reactive oxygen species in H9 c2 cells, and participate in the protection of adriamycin-induced myocardial injury by regulating the NF-κB signaling pathway.

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Objective To study whether intestinal bacterial microenvironment and its metabolites are involved in the occurrence and development of heart failure in rats.Methods The heart failure model of rats was induced by subcutaneous injection of isoproterenol, and the success of the model was evaluated by echocardiography after feeding for one and a half months, and then the rats were divided into two groups: heart failure(HF) group and control group. The serum TMAO level of rats was determined by high performance liquid chromatography tandem mass spectrometry. Fecal samples of the control group and HF group were collected for bacterial DNA extraction,and then conducted 16 SrDNA high-throughput sequencing. The diversity,relative abundance and different species of intestinal flora were analyzed using the NovoMagic cloud platform.Results Compared with the control group,serum TMAO level in HF group significantly increased(P<0. 05). Sequencing results of fecal samples in the two groups showed that at the phylum level,the top three bacteria were Firmicutes,Bacteroidetes and Actinobacteria. At the genus level,the top three species were Lactobacillus,Allobaculum and Duchella. The diversity analysis of the two groups showed that the structure of intestinal flora in HF group was similar to that in control group. The analysis of species differences with the T test showed that the top five bacteria groups with differences between the two groups were Acidobacteriota,Verrucomicrobiota,Nitrospirota,Methylomirabilota and Crenarchaeota(P<0. 05). There was no significant difference in the comparison of other flora(P>0. 05). LEFSE analysis showed that Lactobacillus murinus,Acidobacteriota,Bacteria had significant differences between HF group and control group(LDA absolute value>4).Conclusion The changes of intestinal flora and its metabolites TMAO levels are involved in the occurrence and development of heart failure in rats.

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Objective To explore the effect of Helicobacter pylori(Hp) on the expression of E-cadherin(CDH1) and the expression of CDH1 in gastric cancer tissues, so as to provide a theoretical basis for further exploring CDH1 in the role ofHpin gastric cancer. Methods Mongolian gerbils were gavaged withHp,and their gastric tissues were taken after March, December, and 24 months, and the expression of CDH1 was detected by immunohistochemical staining;Hpinfected gastric epithelial cells AGS,the samples were collected 24 hours later, and Western blot detected each Group CDH1 expression.TCGA and Kaplan-Meier Plotter databases were used to compare the expression differences of CDH1 in gastric cancer tissues and adjacent tissues and the prognostic analysis of CDH1 and gastric cancer; R language was used to perform KEGG enrichment analysis of CDH1 gene; human gastric cancer tissues were collected from the Department of Pathology of your hospital in 10 cases each of the tissue specimens and adjacent tissues, the expression of CDH1 was detected by immunohistochemical staining. Results Immunohistochemical staining results showed that the positive rate of CDH1 was 13.70%,13.20% and 22.90%,respectively, in the stomach tissues of Mongolian gerbils afterHpinfection at 3,12 and 24 months, and 0.04%,4.50% and 4.77%,respectively, in the control group, with statistical significance(P<0.05).The gray values of CDH1 protein expression inHp-infected AGS cells was higher than that in control cells, respectively, with statistical significance(P<0.05). A total of 449 cases with clinicopathological parameters and their corresponding CDH1 mRNA expression level were collected in TCGA gastric cancer database. The CDH1 mRNA expression level in adjacent tissues(76.36±0.48) was significantly lower than that in gastric cancer tissues(147.59±0.11),and the difference was statistically significant(P<0.05).Kaplan-meier Plotter database analysis showed that high expression of CDH1 in gastric cancer decreased the survival rate(P<0.05).KEGG enrichment showed that CDH1 expression was associated with gastric cancer, thyroid cancer, bladder cancer, endometrial cancer, melanoma and other tumors.The function of CDH1 was significantly enriched in bacterial invasion of epithelial cells and adhesion connection, and the pathways were significantly enriched in Apelin signaling pathway, Hippo signaling pathway and Rap1 signaling pathway.The positive rate of CDH1 in human gastric cancer tissues was 76.27%,which was higher than that in para-cancer tissues 45.68%,and the difference was statistically significant(P<0.05). Conclusion Hpinfection causes the increase of CDH1 expression level, which may be one of the reasons thatHpcauses gastric cancer.

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Objective To investigate the effect of HUA on insulin resistance in cardiomyocytes and hyperuricemic mouse model. Methods We exposed primary cardiomyocytes and a rat cardiomyocyte cell line H9 c2 cardiomyocytes to HUA,then quantified glucose uptake with a fluorescent glucose analog, 2-NBDG,after insulin challenge and detected reactive oxygen species(ROS) production.Western blot analysis was used to examine the levels of phosphorylated insulin receptor substrate 1(IRS1,Ser307) and phospho-Akt(Ser473).We monitored the impact of HUA on insulin resistance, insulin signaling, phospho-IRS1(Ser307) and phospho-Akt levels in myocardial tissue of an acute hyperuricemia mouse model established by potassium oxonate treatment.Results HUA inhibited insulin-induced glucose uptake in H9 c2 cardiomyocytes. It increased ROS production,pretreatment with N-acetyl-L-cysteine(NAC),a ROS scavenger,reversed HUA-inhibited glucose uptake induced by insulin. HUA exposure directly increased the phospho-IRS1(Ser307) response to insulin and inhibited that of phospho-Akt in H9 C2 cardiomyocytes,which was blocked by NAC. Furthermore,the acute hyperuricemic mice model showed impaired glucose tolerance and insulin tolerance accompanied by increased phospho-IRS1(Ser307) and inhibited phospho-Akt response to insulin in myocardial tissues.Conclusion HUA inhibits insulin signaling and induces insulin resistance in cardiomyocytes in vitro and in vivo, which is a novel potential mechanism of hyperuricemic-related cardiovascular disease.

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Objective To confirm the therapeutic angiogenesis of danhong injection(DHI) in acute myocardial infarction(AMI) of rats.Methods Forty rats were randomly divided into five groups: Sham group, Model group,DHI low-dose(0. 5 ml/kg) group, mid-dose(1. 0 ml/kg) group, high-dose(2. 0 ml/kg) group. The mouse model of AMI was induced by ligation of the left anterior descending coronary artery. Rats were treated with different doses of DHI(0. 5, 1. 0, 2. 0 ml/kg) for 7 days. Cardiac function of rats was indicated by left ventricular ejection fraction(LVEF) and fractional shortening(LVFS), serum levels of troponin T(TN-T) and brain natriuretic peptide(BNP), myocardial pathological damage, CD34 positive microvessel density(MVD), the expression of Von willebr and factor( vWF), alpha-smooth muscle actin （ α-SMA） and vascular endothelial growth factor-A(VEGF-A) were measured by echocardiography, enzyme linked immunosorbent assay, HE staining, immunohistochemistry and Western blot, respectively.Results Compared with Sham group, LVEF,LVFS in Model group increased(P<0. 01), serum levels of TN-T and BNP decreased(P<0. 01), α-SMA(P<0. 05), CD34,vWF,VEGF-A(P<0. 01) were up-regulated. Compared with Model group, LVEF,LVFS and the expression of CD34,vWF in mid-dose and high-dose groups increased(P<0. 05,P<0. 01). TN-T,BNP in all dosage groups were reduced while the expression of α-SMA,VEGF-A were further upgraded(P<0. 05,P<0. 01). Besides, the pathological damage in the marginal zone of infarcted myocardium was lessened.Conclusion DHI has an effect on therapeutic angiogenesis for AMI in rats.

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Objective To prepare monoclonal antibodies(mAbs) against nucleocapsid protein(N) of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) and identify their biological characteristics. Methods The recombinant SARS-CoV-2 N protein was induced for expression in Escherichia coli.Based on hybridoma technology, SP2/0 mouse myeloma cells were fused with the spleen cells of BALB/c mice immunized with the recombinant protein.The positive hybridoma cell lines were screened by indirect enzyme-linked immunosorbent assay(ELISA) and limiting dilution method.The subtype of mAbs was also identified by indirect ELISA.Then, a large amount of mAbs was produced by ascites injection.After purification with Protein G affinity column, the purity, titer and specificity of mAbs were measured by SDS-PAGE,indirect ELISA and Western blot. Results A total of five hybridoma cell lines that could stably secrete SARS-CoV-2 N protein specific mAbs were obtained and named as N1 to N5.Five mAbs were collected from ascites.Their subtypes were identified as IgG2 a*κ for one mAb and IgG1*κ for the others, and their titers were above 2×104.Western blot showed that the five mAbs could bind to recombinant SARS-CoV-2 N protein. Conclusion Five mAbs that can specifically recognize recombinant SARS-CoV-2 N protein were successfully prepared by hybridoma technology, which lays a foundation for exploration of immunodiagnostic reagents.

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Objective To study the effects of sorafenib on the proliferation, cycle and apoptosis of human intestinal cancer RKO cells. Methods RKO cells were treated with sorafenib at different concentrations(0.5,1.0,5.0,10.0,20.0,40.0 μmol/L),and the inhibitory effect of sorafenib on the proliferation of RKO cells was detected by MTT method, and the half inhibition of concentration(IC50) was determined, set as the experimental group.The control group was treated with dimethyl sulfoxide.For the two groups of cells, the cell clone formation plate method was used to detect the proliferation ability, the flow cytometry was used to detect cell apoptosis and cell cycle changes, and the expression level of intracellular apoptosis-related proteins Caspase-3,Caspase-8 and PARP were detected by Western blot. Results Sorafenib had an inhibitory effect on the proliferation of RKO cells in a concentration-dependent manner.The IC50of sorafenib on RKO cell lines was 10 μmol/L.Compared with the control group, sorafenib(10 μmol/L) could inhibit the clone formation ability of RKO cells(P<0.001) and increase the apoptosis rate[(32.53±6.64)%vs(4.47±2.09)%,P<0.01],affect the cell cycle(P<0.01) and promote the expression levels of apoptosis-related proteins Caspase-3,Caspase-8 and PARP(P<0.05). Conclusion Sorafenib has the ability to inhibit the proliferation of human colorectal cancer RKO cells, and its mechanism may be related to promoting RKO cells apoptosis and affecting the cell cycle.

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Objective To observe the effect of mTOR pathway on glycolysis of T cells after Echinococcus granulosus infection. Methods Balb/c mice were randomly divided into control group, infection group and inhibitor group. In the infection group and inhibitor group, protoscolex was injected under the liver capsule, the control group was injected with an equal volume of PBS, and the inhibitor group was intraperitoneally injected with rapamycin(1.25 mg/kg) daily from the next day. At 15, 30, and 70 days after infection, peripheral blood and spleen were collected, qRT-PCR was used to detect the mRNA expression of metabolic enzymes, and Western blot was used to detect the protein expression of the mTOR signaling pathway. Results Compared with the control group, the protein expression of HIF-1α, P-p70 S6 and P-4 EBP-1 downstream of the mTOR pathway in the infected group decreased(P<0.05), and the mRNA expression of glycolysis-related transcription factors HK3, PKM2, LDHA and MCT4 decreased(P<0.05). After rapamycin inhibited the mTOR pathway, the mRNA expression of glycolysis-related transcription factors HK3, PKM2, LDHA and MCT4 were further reduced(P<0.001). Conclusion Echinococcus infection inhibits glycolytic activity in T cells through the mTOR pathway.

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Objective To analyze the clinical data, biochemical indexes and distribution characteristics of patients with embryonic chromosome abnormalities with early spontaneous abortion, and to explore the risk factors that may cause embryonic chromosomal abnormalities. Methods 315 patients with early spontaneous abortion and chromosome detection of abortion tissue were selected.According to the result of the chromosomes of villi tissue microarray analysis(CMA),315 patients were divided into abnormal group(165 cases) and normal group(150 cases).The clinical data and biochemical indexes of the two groups were analyzed and compared.The screened variables with statistically significant differences were analyzed by multiple Logistic regression. Results The rate of chromosome abnormality in early pregnancy spontaneous abortion embryos was 52.38%.Aneuploid, polyploid, and chromosome structural abnormalities accounted for 71.52%,8.48% and 20.00%,respectively.Compared the abnormal group with the normal group, the distribution of the male age, female nationality, education, career, life history, birth history and previous medical history had no statistically significant differences(P>0.05).There were no statistically significant differences between the two groups in the levels of body mass index(BMI),serum antinuclear antibodies(ANA),anticardiolipin antibody(ACA),antithyroid peroxidase antibody(TPOAb),antithyroglobulin antibody(TgAb),fasting insulin(FINS),fasting blood sugar(FPG),homocysteine(HCY),and sex hormone(P>0.05).Compared the abnormal group with the normal group, the differences of female age, method of conception, serum 25-(OH)D and thyroid stimulating hormone(TSH) had statistical significance(P<0.05).Multivariate analysis showed that female advanced age and vitamin D deficiency were risk factors for fetal chromosomal abnormalities. Conclusion Embryonic chromosome abnormalities are related to female age, method of conception, levels of serum 25-(OH)D and TSH.Female advanced age and vitamin D deficiency may increase the risk of embryonic chromosome abnormalities.

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Objective To quantitatively analyze creeping fat in energy spectrum CT to evaluate the activity of Crohn′s disease. Methods A retrospective study was performed on 127 patients with CD who received energy spectrum CT.The activity of CD was assessed by classical clinical CDAI and these patients were divided into remission group, mild-activity group and moderate and severe-activity group.The slope of spectral HU curve(λHU),normalized iodine concentration(NIC) and normalized water concentration(NWC) of the creeping fat in the diseased intestine were measured and calculated.The visceral fat area(VFA) and subcutaneous fat area(SFA) of all patients were measured by CT post-processing workstation.The relationship between these quantitative data and the inflammatory activity of Crohn′s disease was analyzed. Results The λHU、NIC and NWC in moderate and severe CD patients were higher than those in mild CD patients [λHU(-0.49±0.09)vs(-1.24±0.13),P<0.05;NIC(-4.25±0.96) mg/mlvs(-13.39±2.24) mg/ml,P<0.05;NWC(955.72±5.80) mg/ml vs(931.04±30.25) mg/ml,P<0.05].VFA and VFA/SFA in moderate and severe group were also higher than those in mild group [VFA(79.40±3.58)cm2vs(74.24±6.57)cm2,P<0.05;VFA/SFA(1.01±0.05)vs(0.55±0.09),P<0.05],while SFA in moderate and severe group was significantly lower than that in mild group[(75.57±2.52) cm2vs(137.78±18.31) cm2,P<0.05].λHU,NWC and NIC were all positively correlated with CDAI score(whollyP<0.05),and correlation coefficient r of λHu was 0.881(P<0.001). The AUC of λHU,NIC,NWC and VFA/SFA were respectively 0.957(95%CI:0.922～0.993),0.926(95%CI:0.876～0.977),0.856(95%CI:0.776～0.936),0.793(95%CI:0.699～0.879) in the ROC curve of diagnosing moderate and severe group. Conclusion Energy spectrum CT multi-parameter imaging quantitative analysis of creeping fat is an effective method to evaluate inflammatory activity in patients with CD.

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Objective To study the effect of chronic unpredictable stress(CUS) on the sexual behavior of mice and its possible molecular mechanism. Methods 20 male C57 BL/6 J mice were divided into two groups randomly: control group(n=10) and CUS group(n=10), and the weight of the mice between the two groups was balanced. The control group was 3-5 mice per cage and normal feeding. CUS group was subjected to 21 days CUS combined with single cage fed to establish a depression model. After 21 days, the depressive-like behaviors of the two groups were tested by sucrose preference test and forced swimming test. Then the sexual behavior was detected by olfactory preference test and female urine sniffing test, and the protein and mRNA expression of brain-derived neurotrophic factor(BDNF) in ventral tegmental area(VTA) were detected. Results CUS could significantly decrease sucrose preference(P<0.05) and increase the immobility time in the forced swim test(P<0.05)compared to control mice, indicating depression-like behavior in mice, and impaired sexual behavior in the olfactory preference test(P<0.05)and female urine sniffing test(P<0.05)in mice, and decreased the expression of BDNF in VTA brain regions. The difference was statistically significant. Conclusion The impaired sexual behavior of mice induced by CUS may be related to the decreased expression of BDNF in VTA brain regions. The BDNF signaling pathway may be involved in the regulation of impaired sexual behavior of mice induced by CUS.

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Objective To analyze the influencing factors of patients with tuberculous pleurisy complicated with pleural tuberculoma, and to provide scientific basis for the formulation of treatment plan. Methods Patients with tuberculous pleurisy were selected as the research subjects. All the subjects were divided into case group and control group according to whether they were accompanied by pleural tuberculosis tumor. Self-made questionnaires were used to collect clinical data and laboratory indicators of patients. Results A total of 276 patients with tuberculous pleurisy, with an average age of(39.22±18.12) years, were enrolled in this study, and 84 of them had complicated pleural tuberculoma. Univariate analysis showed that effusion volume, urokinase injection and glucocorticoid use were the influencing factors for the occurrence of pleural tuberculoma in patients with tuberculous pleurisy(P<0.05). Further multivariate analysis showed that pleural effusion content, urokinase injection and glucocorticoid use were statistically significant with the incidence of pleural tuberculoma in patients with tuberculous pleurisy(P=0.003,<0.001,0.001). Conclusion Pleural effusion content, urokinase injection, and the use of glucocorticoid may influence the occurrence of pleural tuberculoma in patients with tuberculous pleurisy.

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Objective To investigate the expression of circFAT1 in non-small cell lung cancer(NSCLC) tissues and the effect of silencing circFAT1 on the proliferation and invasiveness of NSCLC cell A549.Methods A total of107 patients with NSCLC who underwent surgical resection were selected. Real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expressions of circFAT1 in NSCLC and adjacent tissues. A549 cells were cultured and transfected with siRNA-circFAT1(si-circFAT1 group) and irrelevant sequences(si-control group),respectively,and in addition,the untransfected cells were set as the blank group. RT-qPCR was used to detect the expression of circFAT1 in cells,CCK-8 was used to detect cell proliferation,and Transwell chamber was used to detect cell migration and invasion.Results The relative expression level of circFAT1 in NSCLC tissues was higher than that in the adjacent tissues[(2. 47 ± 0. 27)vs(1. 02 ± 0. 15),t= 48. 959,P<0. 001]. Compared with TNM stagesⅠ-Ⅱ,moderately well differentiated and no lymph node metastasis,the relative expression levels of circFAT1 in NSCLC tissues with TNM stages Ⅲ-Ⅳ,poorly differentiated and lymph node metastasis increased(P<0. 05).Compared with the si-control group and the blank group,the relative expression level of circFAT1 in the cells of the si-circFAT1 group was significantly reduced(P<0. 05),and the numbers of migrating cells and invading cells were reduced(P<0. 05).Conclusion The relative expression level of circFAT1 in NSCLC tissues increased. Silencing circFAT1 can inhibit the proliferation,migration and invasion of A549 cells.

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Objective To evaluate the effect and mechanism of the arterial perforators layered skin flap in repair and treatment of various deep surface wounds. Methods 90 patients with deep wounds were randomly divided into the artery perforators layered flap group and traditional flap group, the former were repaired with prominent arterial perforators layered skin flaps, the latte were repaired with traditional flaps.The differences in length of stay, treatment cost, patient satisfaction at discharge, daily living ability, primary wound healing rate, incidence of complications, wound recurrence rate, and total good rate of wound healing between the two groups were observed. Results Compared with the traditional flap group, arterial perforator layered flaps group had advantages in primary wound healing rate, complication rate and total excellent and good rate, and the difference was statistically significant(P<0.05).There was no significant difference in wound recurrence between the two groups(P>0.05).Compared with traditional flap group, the treatment costs, length of hospital stay, living ability score and patient satisfaction of artery perforators layered flap group had advantages with statistically significant difference(P<0.05). Conclusion Arterial perforators layered flap is an ideal measure to repair deep body surface wounds.It is very important to design layered skin flap according to the principle of individualization.Adequate preparation of wounds, exquisite surgical operation and careful management of post-operation are the guarantee of deep wounds' recovery.

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Objective To explore the impact of geriatric nutritional risk index(GNRI) and clinical features on prognosis of patients with diffuse large B-cell lymphoma(DLBCL) based on multicenter data.Methods 505 DLBCL patients diagnosed in 7 medical centers of Huaihai Lymphoma Working group from October 2009 to April 2021 were selected. MaxStat was used to determine the best cut-off value of GNRI. After 1 ∶ 1 propensity score matching(PSM)to balance the covariates between groups,the differences of baseline characteristics and overall survival(OS)between two groups before and after matching were compared. Cox proportional hazards model were used for univariate and multivariate analysis. Kaplan Meier analysis was used to calculate the survival rate and draw the survival curve. Log rank test was used to compare the differences between groups.Results The median age of patients including 273(54. 1%) males was 68 years old. The median survival time was 62. 9 months. Multivariate analysis demonstrated that GNRI,age,LDH and Ann Arbor stage were independent influencing factors of DLBCL(P<0. 05). The best cut-off value of GNRI was 99 based on MaxStat. After PSM,a total of 102 patients in GNRI<99 group could be propensity matched to GNRI ≥99 patients, creating a group of 204 patients. Subgroup analysis showed that GNRI could accurately stratify patients in BCL-2 negative group,BCL-6 positive group,CD5 positive and negative groups,Ann Arbor（Ⅲ-Ⅳ）group and IPI(HIR + HR)group.Conclusion The prognosis of DLBCL is influenced by GNRI,age,LDH and Ann Arbor stage. GNRI has a certain value in evaluating the prognosis of DLBCL patients,and the patients with GNRI≥ 99 indicates a good prognosis.

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Objective To investigate the effects of cognitive behavioral therapy(CBT) combined with SSRI on heart rate variability and vascular endothelial cell function in patients with coronary heart disease(CHD) complicated with anxiety disorder. Methods Ninety patients with CHD combined with anxiety disorder were randomly divided into observation group(CBT combined with SSRI treatment) and control group(SSRI treatment). The changes of 17 items of Hamilton Depression Scale(HAMD-17) score, Hamilton Anxiety Scale(HAMA) score, heart rate variability and vascular endothelial cell function were compared between the two groups before and after treatment. Results After treatment, HAMD-17 and HAMA in both groups were lower than those before treatment, while HAMD-17 and HAMA in observation group were lower than those in control group(P<0.05). After treatment, ET-1, IL-6, TNF-α and vWF decreased in the two groups, while NO increased(P<0.05). After treatment, ET-1 and vWF in the observation group were lower than those in the control group, while NO was higher than that in the control group(P<0.05). After treatment, SNDD, RMSSD, SDANN and PNN50 in the observation group were higher than those before treatment(P<0.05), and all indexes in the control group also showed an increasing trend, but the differences were not statistically significant. After treatment, SNDD and RMSSD in observation group were higher than those in control group(P<0.05). Conclusion CBT combined with SSRI can improve anxiety symptoms, autonomic nerve function and endothelial cell function in CHD patients.

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Objective To screen the differentially expressed mRNA between esophageal squamous cell carcinoma(ESCC) and normal tissues of Kazakh in Xinjiang by microarray, so as to further understand the occurrence and development mechanism of ESCC. Methods Five pairs of esophageal squamous cell carcinoma and adjacent normal tissues(more than 5 cm away from cancer tissues) were screened by gene chip; Bioinformatics analysis was performed by R software and R software package; Metascape was used to carry out gene ontology(GO) enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analyses; bioinformatics website String and Cytoscape software was used to find core genes; 23 cases of esophageal squamous cell carcinoma(ESCC) and 20 cases of adjacent normal tissues of Kazakh in Xinjiang were collected, and the core gene PLAUR was verified by immunohistochemistry. Results 1 764 differentially expressed mRNAs were screened out(multiple of difference ≥2 andP<0.01); Among the differentially expressed mRNAs, 378 genes were up-regulated and 1 368 genes were down regulated.These differentially expressed mRNAs involved a series of biological functions and pathways; Immunohistochemical results showed that the expression of the core gene PLAUR in Kazakh esophageal squamous cell carcinoma was higher than that in adjacent normal tissues. Conclusion The differentially expressed mRNAs in esophageal squamous cell carcinoma and adjacent normal tissues of Kazakh in Xinjiang was screened by microarray, and the core genes were found out, which improved the gene spectrum of esophageal squamous cell carcinoma of Kazakh in Xinjiang, and laid a foundation for further research on the diagnosis and prognosis of esophageal squamous cell carcinoma.

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Objective The study aimed at exploring the expression level and clinical significance of miR-22 in serum of patients with non-small cell lung cancer. Methods 276 patients with NSCLC diagnosed were collected.The expression of miR-22 in serum was detected by qRT-PCR,and its correlation with clinical features and prognosis was analyzed. Results The serum level of miR-22 in NSCLC patients was higher than that in normal subjects(P<0.01).According to the different clinical characteristics of NSCLC patients, we could know that the expression of miR-22 was different in M stage, N stage, tumor differentiation degree, lung cancer bone metastasis, lung cancer liver metastasis and intrapulmonary metastasis groups(P<0.05).High level group and low level group were divided according to the average level of serum miR-22 in NSCLC patients, and the expression of serum miR-22 was correlated with N stage, M stage, liver metastasis and intrapulmonary metastasis of lung cancer.Survival analysis showed that the prognosis of NSCLC patients with low serum miR-22 expression was better than that of the patients with high serum miR-22 expression(P<0.05).Multivariate Cox analysis confirmed that serum miR-22,T stage, M stage, bone metastasis of lung cancer and primary tumor laterality were independent prognostic factors of NSCLC patients. Conclusion The expression of serum miR-22 is closely related to the occurrence and development of non-small cell lung cancer patients, which can be used as an independent prognostic factor for NSCLC patients.

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Peripheral blood of healthy people was used to induce the proliferation and differentiation of B lymphocytes into plasmablasts and plasma cells in vitro.Cells characteristics were observed under microscope.Molecules expression level of B cells surface was analyzed under flow cytometry.Cell viability was observed by Trypan blue.The result showed CD40 L signal stimulation, CpG immunoregulatory sequence combination and optimized cytokine combination could effectively promote the transformation of human B lymphocytes and the establishment of vitro model.

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To evaluate the application value of 2020 Chinese guidelines for ultrasound malignancy risk stratification of thyroid nodules( the C-TIRADS) in the diagnosis and follow-up management of thyroid nodules.The study included the 251 thyroid nodules in 208 patients, and ultrasonographic features of each nodule were assessed according to the criteria of C-TIRADS.The area under ROC curve was 0.942(95%CI0.914-0.971), when 4 B was set as the cut-off value of benign and malignant nodules, the diagnostic efficacy was highest, with the sensitivity of 0.916, specificity of 0.849, accuracy of 0.888 and Youden index of 0.765.C-TIRADS is of great value in guiding the diagnosis and treatment of thyroid nodules.