Objective To investigate the effect of programmed cell death-ligand 1(PD-L1)monoclonal antibody and paclitaxel(PTX)combined with lentinan(LNT)on human breast cancer MDA-MB-231 cells in vitro.Methods MDA-MB-231 cells,human peripheral blood mononuclear cells(PBMCs),and MDA-MB-231+PBMCs co-culture were randomly divided into control group,PTX group,LNT group,MPDL3280A(PD-L1 monoclonal antibody)group,PTX+LNT group,and PTX+LNT+MPDL3280A group.The cell activity was detected by CCK8.The expressions of MHC-I and PD-L1 were detected by flow cytometry.The levels of IFN-γ and TNF-α were detected by ELISA kits.Results Compared with control group,the activity of MDA-MB-231 cells was significant inhibited in PTX group,MPDL3280A group,PTX+LNT group,and PTX+LNT+MPDL3280A group(P＜0.01).LNT group and PTX+LNT+MPDL3280A group significantly promoted the immune effect of PBMCs(P＜0.05,P＜0.01).PTX+LNT+MPDL3280A group significantly inhibited the activity of MDA-MB-231cells co-cultured with PBMCs(0.56±0.16 vs.0.39±0.13,P＜0.05).Compared with the negative control,LNT significantly promoted the expression of PD-L1 in MDA-MB-231 and the secretion of IFN-γ by PBMCs(P＜0.05).Conclusion By blocking the effect between PD-L1 and PD-1,PD-L1 monoclonal antibody can improve immunity and promote the antitumor effect of PTX combined with LNT in vitro.

Objective To investigate the effect of programmed cell death-ligand 1(PD-L1)monoclonal antibody and paclitaxel(PTX)combined with lentinan(LNT)on human breast cancer MDA-MB-231 cells in vitro.Methods MDA-MB-231 cells,human peripheral blood mononuclear cells(PBMCs),and MDA-MB-231+PBMCs co-culture were randomly divided into control group,PTX group,LNT group,MPDL3280A(PD-L1 monoclonal antibody)group,PTX+LNT group,and PTX+LNT+MPDL3280A group.The cell activity was detected by CCK8.The expressions of MHC-I and PD-L1 were detected by flow cytometry.The levels of IFN-γ and TNF-α were detected by ELISA kits.Results Compared with control group,the activity of MDA-MB-231 cells was significant inhibited in PTX group,MPDL3280A group,PTX+LNT group,and PTX+LNT+MPDL3280A group(P＜0.01).LNT group and PTX+LNT+MPDL3280A group significantly promoted the immune effect of PBMCs(P＜0.05,P＜0.01).PTX+LNT+MPDL3280A group significantly inhibited the activity of MDA-MB-231cells co-cultured with PBMCs(0.56±0.16 vs.0.39±0.13,P＜0.05).Compared with the negative control,LNT significantly promoted the expression of PD-L1 in MDA-MB-231 and the secretion of IFN-γ by PBMCs(P＜0.05).Conclusion By blocking the effect between PD-L1 and PD-1,PD-L1 monoclonal antibody can improve immunity and promote the antitumor effect of PTX combined with LNT in vitro.

This study was designed to investigate the biological function and interaction mechanism of deubiquitinating enzyme USP14 and β-catenin in endometrial carcinoma(EC).The expression of USP14 and β-catenin in EC tissues and adjacent healthy tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry,while the effects of USP14 on β-catenin protein stability and ubiquitination were investigated by Western blotting and protein immunoprecipitation using HEA-1C and Ishikawa cell lines derived from EC.The effect of USP14 and β-catenin on EC cell proliferation was verified by cell proliferation assay(CCK-8);the regulatory effect of USP14 on β-catenin was verified by plasmid transfection.The regulation of USP14 and β-catenin on the expression of proteins related to epithelial-mesenchymal transition(EMT)in EC cells were verified by Western blotting.Data showed that the expression of USP14 and β-catenin in EC tissues were higher than those in adjacent healthy tissues,and USP14 and β-catenin interacted in EC cell line.USP14 maintained the protein stability of β-catenin through its deubiquitination activity.USP14-specific inhibitors could enhance the ubiquitination level of β-catenin and down-regulate the expression of β-catenin.USP14-specific inhibitors also inhibited EC cell proliferation and inhibited the expression of EMT-related proteins,while overexpression of β-catenin reversed the inhibition of EC cell growth and EMT induced by USP14 inhibitors.In conclusion,USP14 maintains β-catenin protein stability in EC cells by regulating the ubiquitination level,and the regulation of cell proliferation and EMT requires β-catenin.

This study was designed to investigate the biological function and interaction mechanism of deubiquitinating enzyme USP14 and β-catenin in endometrial carcinoma(EC).The expression of USP14 and β-catenin in EC tissues and adjacent healthy tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry,while the effects of USP14 on β-catenin protein stability and ubiquitination were investigated by Western blotting and protein immunoprecipitation using HEA-1C and Ishikawa cell lines derived from EC.The effect of USP14 and β-catenin on EC cell proliferation was verified by cell proliferation assay(CCK-8);the regulatory effect of USP14 on β-catenin was verified by plasmid transfection.The regulation of USP14 and β-catenin on the expression of proteins related to epithelial-mesenchymal transition(EMT)in EC cells were verified by Western blotting.Data showed that the expression of USP14 and β-catenin in EC tissues were higher than those in adjacent healthy tissues,and USP14 and β-catenin interacted in EC cell line.USP14 maintained the protein stability of β-catenin through its deubiquitination activity.USP14-specific inhibitors could enhance the ubiquitination level of β-catenin and down-regulate the expression of β-catenin.USP14-specific inhibitors also inhibited EC cell proliferation and inhibited the expression of EMT-related proteins,while overexpression of β-catenin reversed the inhibition of EC cell growth and EMT induced by USP14 inhibitors.In conclusion,USP14 maintains β-catenin protein stability in EC cells by regulating the ubiquitination level,and the regulation of cell proliferation and EMT requires β-catenin.

Objective To understand the endemic situation and control effect of schistosomiasis through the surveillance in a national surveillance site of Jurong City, so as to provide the evidence for formulating the prevention and control measures. Methods According to the National Schistosomiasis Monitoring Scheme (2014 Edition), the surveillance of schistosome infection in Oncomelania hupensis snails, residents and livestock was performed in the Kongqing Village, a national surveillance site of Jurong City, from 2015 to 2017. Results The areas with snails were 0, 0, and 0.63 hm² in 2015, 2016, and 2017 respectively; the average densities of living snails were 0, 0, and 0.19 snails/0.1 m² in 2015, 2016, and 2017 respectively. No schistosome-infected snails were found. The positive rates of blood tests for schistosomiasis in the local residents were 7.72%, 7.45% and 3.45%, and the positive rates of blood tests in the floating population were 4.90%, 3.47% and 0.97% in 2015, 2016 and 2017, respectively. No positives were found in the schistosome etiology detection in the crowd and livestock. Conclusions The effect of schistosomiasis prevention and control is obvious in Jurong City, but O. hupensis snails are still of recurrence. Therefore, the monitoring and control efforts should be strengthened to consolidate the achievements of schistosomiasis prevention and control.

Objective To characterize the biomarkers of urine samples for early diagnosis of colorectal cancer(CRC)using proton nuclear magnetic resonance (1H-NMR)combined with pattern recognition.Methods 400 MHz 1H-NMR was used to test the urine samples obtained from 23 patients with Ⅰ/Ⅱ stage CRC,40 healthy controls (HC)and 18 patients with esophageal cancer (EC). Pattern recognition through orthogonal partial least squares-discriminant analysis (OPLS-DA)was applied on 1H-NMR data to find urine metabolic differences between CRC and HC.Results OPLS-DA could effectively determine HC,patients withⅠ/Ⅱstage CRC and patients with esophageal cancer.Compared with HC,early stage CRC had significant decreases of choline,isocitric acid,lactamine,phenylalan, cysteine,creatinine,aspartic acid,hippurate acid,methylamine,dimethyl sulfone,and increases of acetoacetate,glutamine,glycocyamine,cis-aconitate, trans-aconitate,homocycteine in the urine samples.Conclusion Urine metabonomics based on NMRIndicates that glucose metabolism,amino acid metabolism,choline metabolism,energy metabolism and intestinal microflora are disturbance in colorectal cancer patients,which provide valuable metabolic information on the molecular level for early diagnosis of colorectal cancer.

Objective To investigate the effects of metformin(Met)on liver function,chronic inflammation and liver fibrosis in rats with type 2 diabetes mellitus (T2DM) combined with nonalcoholic fatty liver disease (NAFLD). Methods 4 groups of rat models were established:the control group(group A),T2DM group(group B),T2DM+NAFLD group(group C)and T2DM+NAFLD+Met group(group D). After grouping,the specimens of blood and liver tissues were taken. Serum liver function,C-reactive protein(CRP),tumor necrosis factor-α (TNF-α)and interleukin-6(IL-6)were detected,and then their insulin resistance was assessed. The pathological changes of the liver were detected by HE staining,oil red O staining,Masson staining and reticular fiber staining, and the expression of TNF-α,IL-6 and CRP in liver tissue was detected by immunohistochemistry. Results (1) The degree of insulin resistance and hypohepatia in group B and C were higher than those in group A,and those of group D were lower than those in group C.(2)The levels of inflammatory cytokines in group B and C were higher than those in group A,and the level in group D were lower than those in group C.(3)The pathological examination of liver showed that there was significant fatty degeneration,fatty inflammation and fibrosis in the livers of group C,and group D was better than those in group C. Conclusions Met is effective in the improvement of insulin resis-tance,hypohepatia,chronic inflammation and liver fibrosis in type 2 diabetes mellitus associated with non-alco-holic fatty liver disease,which in mechanism may be related to the decreased expression of inflammatory factors and improved chronic inflammatory state.

AIM:To investigate the effect of insulin and selenium in combination on the apoptosis and the ex-pression of Ku70, acetylated Ku70, Bax and cytochrome C in myocardial cells of the rats with diabetic cardiomyopathy (DCM), and to explore the mechanism of insulin and selenium in their synergistic anti-DCM effect.METHODS:SD rats (n=50) were randomly grouped into control, DCM, DCM with insulin treatment (DCM+In) group, DCM with selenium treatment (DCM+Se) group, and DCM with insulin and selenium combination treatment (DCM+In+Se) group.Mito-chondrial membrane potential ( MMP) was measured by flow cytometry .The cell apoptosis was observed by terminal-deoxy-nucleotidyl transferase mediated nick end labeling (TUNEL).The levels of Ku70, Bax and cytochrome C were examined by Western blot .The acetylation status of Ku 70 was detected by co-immunoprecipitation .RESULTS: The rats in DCM group showed marked cell apoptosis compared with the control rats .The levels of Ku70 and acetylated Ku70 declined sig-nificantly compared with control group .Bax significantly translocated from cytoplasm into mitochondria and cytochrome C translocated from mitochondria into cytoplasm compared with control group .Compared with DCM +In group or DCM +Se group, insulin and selenium in combination significantly inhibited the apoptosis , down-regulated Ku70 and acetylated Ku70 levels, and prevented Bax and cytochrome C translocation .CONCLUSION: Insulin and selenium synergistically inhibits myocardial apoptosis by regulating Ku 70 acetylation and inhibiting Bax translocation .

Aim To explore the mechanism of insulin in combination with selenium preventing myocardial apoptosis in diabetic cardiomyopathy rats .Methods SD rats ( n =50 ) were randomly divided into five groups: control , diabetic cardiomyopathy ( DCM ) , DCM with insulin treatment , DCM with selenium treat-ment, and DCM with insulin and selenium combination treatment .The cell apoptosis was observed by TUNEL . The levels of Bcl-2, caspase-3, PARP, Cbl-b and p38 MAPK were examined by Western blot .The inter-actions of Cbl-b-p38 MAPK and Ku70-Bax were detec-ted by immunoprecipitation .Results Insulin in com-bination with selenium synergistically inhibited apopto-sis, up-regulated Cbl-b, down-regulated p38MAPK ex-pressions and increased the interactions of Cbl-b-p38MAPK and Ku70-Bax.Conclusion Insulin and selenium synergistically inhibit myocardial apoptosis by regulating Cbl-b-inhibited p38 MAPK and preventing Bax translocation .

Objective To characterize the metabolic "fingerprint" of fecal extracts for diagnosis of early-stage colorectal cancer (CRC) using proton nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomics coupled with pattern recognition.Methods From January 2014 to December 2014,we collected fecal samples at the Second Affiliated Hospital of Shantou University Medical College,from 25 patients with colorectal adenomas (CR-Ad),20 with stage Ⅰ /Ⅱ CRC,and 32 healthy controls (HCs).The patients were diagnosed by histopathology.No subjects had any complicating diseases.HCs showed no abnormalities from blood tests,endoscopic examination,diagnostic imaging,and/or medical interviews.We excluded participants who used antibiotics,NSAIDS,statins,or probiotics within two months of study participation,and any patients who underwent chemotherapy or radiation treatments prior to surgery.We used orthogonal partial least-squares-discriminant analysis (OPLS-DA) for pattern recognition (dimension reduction) on 1H-NMR processed data (1H frequency of 400.13 MHz),to find metabolic differences among CR-Ad,carcinoma and HC fecal samples;and receiver operating characteristic (ROC) analysis to determine the diagnostic value of the fecal metabolic biomarkers.Results Fecal samples were collected from 20 patients with Stage Ⅰ/Ⅱ CRC (11 M,9 F,median age (52±13) years),25 with CR-Ad (14 M,11 F,median age (53±11) years) and 32 HCs (15 M,17 F,median age (53± 14) years).OPLS-DA clearly distinguished CR-Ad and stage Ⅰ/Ⅱ CRC from HC samples,based on their metabolomic profiles.Relative signal intensities in HCs were significantly lower than in the cancer patients for butyrate (HC:23.0±6.0;CR-Ad:18.0±5.0;CRC:14.0±6.0;Z=-2.07,P=0.008),acetate (HC:45.0± 11.0;CR-Ad:31.0±11.0;CRC:24.0±8.0;Z=-2.32,P=0.011),propionate (HC:26.0 ± 7.0;CR-Ad:22.0 ± 6.0;CRC:19.0 ± 5.0;Z=-2.43,P=0.032),glucose (HC:37.0±7.0;CR-Ad:31.0±7.0;CRC:26.0±8.0;Z=-2.07,P=0.044) and glutamine (HC:4.5±2.0;CR-Ad:4.9 ± 1.0;CRC:5.4 ± 1.0;Z=2.21,P=0.044).However,relative signal intensities in HCs were significantly higher than in patients for lactate (HC:4.8± 1.0;CR-Ad:6.9±2.0;CRC:4.8± 1.0;Z=2.02,P=0.038),glutamate (HC:3.2±2.0;CR-Ad:4.9 ± 1.0;CRC:3.2± 2.0;Z=2.21,P=0.044) and succinate (HC:12.0±2.0;CR-Ad:15.0±3.0;CRC:12.0± 2.0;Z=2.25,P=0.011).Among the potential biomarkers,acetate at 1.92 ppm,and succinate at 2.41 ppm displayed relatively high area under ROC,with sensitivity and specificity both ＞90％,to distinguish early-stage CRC patients from HCs.Conclusion Fecal metabolic profiles distinguish of HCs from patients with CRC patients,even in the early stages (stage Ⅰ/Ⅱ),highlighting the potential of NMR-based fecal metabolomic fingerprinting as tools for early CRC diagnosis.