Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.010
Xuzhi Zhang, Ao Ren, Zhongqiu Li, R. Deng, Yi Ma
Objective To study the effect of inhibiting NLRC5 on CD4+ T cells function and allografts immune protective effects in mice. Methods Mice spleen-derived CD4+ T cells were cultured, purified andtransfected with short hairpin RNA (shRNA) lentiviral vector NLRC5-RNA interference (RNAi)-green fluorescent protein (GFP). The mice allogenic islet transplantation model andskintransplantation model were established, and 24 h prior to transplantation each recipient mice was given CD4+ T cells, NLRC5-RNAi-T cells or equal amount of phosphate buffer (PBS), named T cell group, NLRC5-RNAi group and control group respectively. The survival of islet grafts andskingraftsof transplanted recipients was observed, and on the day 7, the islet graft glucose tolerance, the percentage of T cell subsets in the spleenaswellas related cytokines interleukin (IL)-10 and interferon (IFN)-γwere observed. Results The experimental data demonstratedthat NLRC5-RNAi significantly prolonged the survival timewith a median survival time of (19.0±3.4) days inislet allograftand (15.4±3.8) days inskin allograft, which is significantly longer than control [(11.6±1.9) d, t=5.156, P<0.01] and T cell group [(10.0±1.4) d, t=4.151, P<0.01] inislet allograft, aswellascontrol [(8.0±0.9) d, t=3.375, P<0.05] and T cell group [(7.8±0.8) d, t=3.375, P<0.05] inskin allograft. The results of enzyme linked immunosorbent assay (ELISA) showed that the expression of IL-10 (344.0±4.1) ng/L inisletallograft and (275.9±12.5) ng/L in skinallograftin NLRC5-RNAi group were significantly higher than control group (t=124.141, 20.121, P<0.01) and T cell group (t=32.605, 17.900, P<0.01); the expression of IFN-γ (85.1±6.6) ng/L inisletallograftand (96.6±2.5) ng/L inskinallograftin NLRC5-RNAi group were lower than control group (t=7.633, 7.490, P<0.05) and T cell group (t=10.972, 13.286, P<0.01). Flow cytometric analysis revealed that compared to control group and T cell group, the NLRC5-RNAi group dramatically increased the population of Th2 [(0.190±0.053)%, t=5.220, 5.278, P<0.05] inislet allograft and [(0.130±0.012)%, t=21.060, 9.470, P<0.05] in skin allograft, as well asreduced the population of Th1 [(0.810±0.036)%, t=6.219, 5.276, P<0.05] inislet allograft and [(0.180±0.026)%, t=9.248, 25.324, P<0.05] in skin allograft. Conclusion After inhibiting the expression of NLRC5 gene, the expression of Th2 type cytokines in CD4+ T cells is increased, which prolongs the survival time of grafts and has positive significance for the induction of transplantation tolerance. Key words: NLRC5; Lentivirus transduction; CD4+ T cells; Islet transplantation; Skin transplantation; Immune tolerance
{"title":"Inhibit NLRC5 regulate CD4+ T cellsfunctionandimmunological protection onallograftinmice","authors":"Xuzhi Zhang, Ao Ren, Zhongqiu Li, R. Deng, Yi Ma","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.010","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.010","url":null,"abstract":"Objective \u0000To study the effect of inhibiting NLRC5 on CD4+ T cells function and allografts immune protective effects in mice. \u0000 \u0000 \u0000Methods \u0000Mice spleen-derived CD4+ T cells were cultured, purified andtransfected with short hairpin RNA (shRNA) lentiviral vector NLRC5-RNA interference (RNAi)-green fluorescent protein (GFP). The mice allogenic islet transplantation model andskintransplantation model were established, and 24 h prior to transplantation each recipient mice was given CD4+ T cells, NLRC5-RNAi-T cells or equal amount of phosphate buffer (PBS), named T cell group, NLRC5-RNAi group and control group respectively. The survival of islet grafts andskingraftsof transplanted recipients was observed, and on the day 7, the islet graft glucose tolerance, the percentage of T cell subsets in the spleenaswellas related cytokines interleukin (IL)-10 and interferon (IFN)-γwere observed. \u0000 \u0000 \u0000Results \u0000The experimental data demonstratedthat NLRC5-RNAi significantly prolonged the survival timewith a median survival time of (19.0±3.4) days inislet allograftand (15.4±3.8) days inskin allograft, which is significantly longer than control [(11.6±1.9) d, t=5.156, P<0.01] and T cell group [(10.0±1.4) d, t=4.151, P<0.01] inislet allograft, aswellascontrol [(8.0±0.9) d, t=3.375, P<0.05] and T cell group [(7.8±0.8) d, t=3.375, P<0.05] inskin allograft. The results of enzyme linked immunosorbent assay (ELISA) showed that the expression of IL-10 (344.0±4.1) ng/L inisletallograft and (275.9±12.5) ng/L in skinallograftin NLRC5-RNAi group were significantly higher than control group (t=124.141, 20.121, P<0.01) and T cell group (t=32.605, 17.900, P<0.01); the expression of IFN-γ (85.1±6.6) ng/L inisletallograftand (96.6±2.5) ng/L inskinallograftin NLRC5-RNAi group were lower than control group (t=7.633, 7.490, P<0.05) and T cell group (t=10.972, 13.286, P<0.01). Flow cytometric analysis revealed that compared to control group and T cell group, the NLRC5-RNAi group dramatically increased the population of Th2 [(0.190±0.053)%, t=5.220, 5.278, P<0.05] inislet allograft and [(0.130±0.012)%, t=21.060, 9.470, P<0.05] in skin allograft, as well asreduced the population of Th1 [(0.810±0.036)%, t=6.219, 5.276, P<0.05] inislet allograft and [(0.180±0.026)%, t=9.248, 25.324, P<0.05] in skin allograft. \u0000 \u0000 \u0000Conclusion \u0000After inhibiting the expression of NLRC5 gene, the expression of Th2 type cytokines in CD4+ T cells is increased, which prolongs the survival time of grafts and has positive significance for the induction of transplantation tolerance. \u0000 \u0000 \u0000Key words: \u0000NLRC5; Lentivirus transduction; CD4+ T cells; Islet transplantation; Skin transplantation; Immune tolerance","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"33-36"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45494035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.044
W. Bu, Qinglin Zhao, D. Zhao
Objective To study the clinical significance of early usage of continuous blood purification on the treatment of severe craniocerebral injury. Methods 85 Patients with severe brain injury admitted in our department were randomly divided into continuous blood purification treatment group (experiment group, 42 patients) and conventional treatment group (control group, 43 patients). patients in continuous blood purification treatment group were treated by continuous blood purification and traditional treatment from 1-7 days, traditional treatment group only were treated by the traditional treatment methods. To detect tumor necrosis factor (TNF)-α, neuron specific enolase (NSE) and pNF-H in 1 and 5 day. On the 28th day Apache Ⅱ score, ICU stay time and GCS score of two groups. To detect 6 months survival rate of two groups. Spss15.0 software was used for statistical analysis, The measurement data are subject to T test The survival rate and quality of life comparison between the two groups was compared by χ2 test, P<0.05 was considered statistically significant. Results Incidence of 5 days of TNF-α (4.85±0.45) ng/L (t=6.890, P 0.05). Conclusion Early continuous blood purification treatment improves the survival rate and quality of life of patients with severe brain injury which have higher clinical value. Key words: Continuous blood purification; Sever brain injury; Early treatment
{"title":"Clinical study on early application of continuous blood purification in the treatment of severe craniocerebral injury","authors":"W. Bu, Qinglin Zhao, D. Zhao","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.044","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.044","url":null,"abstract":"Objective \u0000To study the clinical significance of early usage of continuous blood purification on the treatment of severe craniocerebral injury. \u0000 \u0000 \u0000Methods \u000085 Patients with severe brain injury admitted in our department were randomly divided into continuous blood purification treatment group (experiment group, 42 patients) and conventional treatment group (control group, 43 patients). patients in continuous blood purification treatment group were treated by continuous blood purification and traditional treatment from 1-7 days, traditional treatment group only were treated by the traditional treatment methods. To detect tumor necrosis factor (TNF)-α, neuron specific enolase (NSE) and pNF-H in 1 and 5 day. On the 28th day Apache Ⅱ score, ICU stay time and GCS score of two groups. To detect 6 months survival rate of two groups. Spss15.0 software was used for statistical analysis, The measurement data are subject to T test The survival rate and quality of life comparison between the two groups was compared by χ2 test, P<0.05 was considered statistically significant. \u0000 \u0000 \u0000Results \u0000Incidence of 5 days of TNF-α (4.85±0.45) ng/L (t=6.890, P 0.05). \u0000 \u0000 \u0000Conclusion \u0000Early continuous blood purification treatment improves the survival rate and quality of life of patients with severe brain injury which have higher clinical value. \u0000 \u0000 \u0000Key words: \u0000Continuous blood purification; Sever brain injury; Early treatment","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"152-154"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48957098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effects of astragaloside IV and 5-fluorouracilon on chemosensitization in SW480 tumor transplanted in nude mice and the associated mechanism","authors":"Jianbin Chen, Fang Wang, Weiwei Wang, Guoling Yin, Jianjun Wu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.051","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.051","url":null,"abstract":"","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"176-176"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42996084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.017
Qinghua Zhang, Gui-Zhen Ou, Ruihua Wang, Zhenxing Yu, Jingfeng Liu
Objective To investigate the effect of common bile duct ligation on fibroblast activation in rats with obstructive jaundice. Methods Twenty-four male specific pathogen free (SPF) rats were randomly divided into sham-operated group and experimental group, with 12 rats in each group. All rats underwent common bile duct ligation. Rats The common bile duct in the experimental group was ligated with 3-0 silk thread and cut off. The rats in the two groups were sampled 2 and 7 days after operation. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of transforming growth factor-β1 (TGF-β1). Immunohistochemistry was used to detect the expression of TGF-β1 protein. The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of TGF-β1 gene. Western blotting was used to detect the expression of TGF-β1 protein. Results In sham-operated group, the structure of hepatic lobules was intact, and the hepatic sinuses were normal without congestion. In experimental group, the structure of hepatic lobules was disordered and the hepatic cords were dissociated. The level of serum TGF-β1 in the experimental group at 2nd day [(689.42±58.13) ng/L] and 7th day [(867.53±45.62) ng/L] was significantly higher than that in the sham-operated group [(519.92±32.41) ng/L and (536.50±41.29) ng/L, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 protein in liver tissues of experimental group at 2nd day [(6.53±1.29)%] and 7th day [(8.97±1.85)%] was significantly higher than that of sham-operated group [(1.17±0.24)% and (1.54±0.39)%, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 in liver tissues of experimental group at 2nd day [(0.89±0.14)%] and 7th day [(0.95±0.19)%] was significantly higher than that of sham-operated group [(0.52±1.32)% and (0.31±1.76)%, t=14.693 and 18.316, P<0.05]. The gray value of TGF-β1 protein expression in liver tissue of experimental group at 2nd day after operation (0.62±0.13) and 7th day after operation (0.78±0.10) was significantly higher than that of sham-operated group (0.21±0.05 and 0.23±0.06, t=10.197 and 16.337, P<0.05). Conclusion The expression of TGF-β1 in rats with obstructive jaundice induced by common bile duct ligation is significantly up-regulated, and the proliferation of biliary duct epithelial cells induces fibroblast activation. Key words: Common bile duct ligation; Obstructive jaundice; Fibroblast activation
{"title":"Fibroblast activation induced by common bile duct ligation in rats with obstructive jaundice","authors":"Qinghua Zhang, Gui-Zhen Ou, Ruihua Wang, Zhenxing Yu, Jingfeng Liu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.017","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.017","url":null,"abstract":"Objective \u0000To investigate the effect of common bile duct ligation on fibroblast activation in rats with obstructive jaundice. \u0000 \u0000 \u0000Methods \u0000Twenty-four male specific pathogen free (SPF) rats were randomly divided into sham-operated group and experimental group, with 12 rats in each group. All rats underwent common bile duct ligation. Rats The common bile duct in the experimental group was ligated with 3-0 silk thread and cut off. The rats in the two groups were sampled 2 and 7 days after operation. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of transforming growth factor-β1 (TGF-β1). Immunohistochemistry was used to detect the expression of TGF-β1 protein. The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of TGF-β1 gene. Western blotting was used to detect the expression of TGF-β1 protein. \u0000 \u0000 \u0000Results \u0000In sham-operated group, the structure of hepatic lobules was intact, and the hepatic sinuses were normal without congestion. In experimental group, the structure of hepatic lobules was disordered and the hepatic cords were dissociated. The level of serum TGF-β1 in the experimental group at 2nd day [(689.42±58.13) ng/L] and 7th day [(867.53±45.62) ng/L] was significantly higher than that in the sham-operated group [(519.92±32.41) ng/L and (536.50±41.29) ng/L, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 protein in liver tissues of experimental group at 2nd day [(6.53±1.29)%] and 7th day [(8.97±1.85)%] was significantly higher than that of sham-operated group [(1.17±0.24)% and (1.54±0.39)%, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 in liver tissues of experimental group at 2nd day [(0.89±0.14)%] and 7th day [(0.95±0.19)%] was significantly higher than that of sham-operated group [(0.52±1.32)% and (0.31±1.76)%, t=14.693 and 18.316, P<0.05]. The gray value of TGF-β1 protein expression in liver tissue of experimental group at 2nd day after operation (0.62±0.13) and 7th day after operation (0.78±0.10) was significantly higher than that of sham-operated group (0.21±0.05 and 0.23±0.06, t=10.197 and 16.337, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The expression of TGF-β1 in rats with obstructive jaundice induced by common bile duct ligation is significantly up-regulated, and the proliferation of biliary duct epithelial cells induces fibroblast activation. \u0000 \u0000 \u0000Key words: \u0000Common bile duct ligation; Obstructive jaundice; Fibroblast activation","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"60-62"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47648342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.030
Ximeng Ji, H. Zhuang, Yiwen Zhang, Jincan Zhang, Bu Shoushan
Objective To construct an artificial nerve graft as an alternative for autologous nerve grafting. Methods We fabricated an innovative tissue engineered nerve graft (TENG), which consists of a commercial bilayer collagen membrane (Bio-Gide)-based nerve scaffold and bone marrow mesenchymal stem cells (BMSCs) serving as support cells to bridge a 10 mm long sciatic nerve defect in rats, The 10 mm gap was left empty after resection in the blank group. In the other three groups, the sciatic nerve defect was, respectively, bridged with plain collagen tube (CT group), a collagen tube filled with BMSC suspension (C+ CT group), or reversed transected nerve segment (AG group). Results Histological and functional assessments showed that the developed TENG facilitated nerve regeneration (SFI: CT, -92.490±1.836; C+ CT, -70.010±7.805; AG, -70.130±8.744; F= 17.850, P 0.05) and better outcomes than the plain collagen tube (P<0.05). Conclusion These results manifested that the bilayer collagen conduit loaded BMSCs could be applied as a new biodegradable artificial nerve guide for nerve tissue engineering. Key words: Tissue engineer; Nerve graft; Bone marrow mesenchymal stem cells; Bilayer collagen conduit scaffold; Sciatic nerve injury; Peripheral nerve regeneration
{"title":"Repairing rat sciatic nerve defects via transplanting bilayer collagen conduit scaffolds carrying bone marrow mesenchymal stem cells","authors":"Ximeng Ji, H. Zhuang, Yiwen Zhang, Jincan Zhang, Bu Shoushan","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.030","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.030","url":null,"abstract":"Objective \u0000To construct an artificial nerve graft as an alternative for autologous nerve grafting. \u0000 \u0000 \u0000Methods \u0000We fabricated an innovative tissue engineered nerve graft (TENG), which consists of a commercial bilayer collagen membrane (Bio-Gide)-based nerve scaffold and bone marrow mesenchymal stem cells (BMSCs) serving as support cells to bridge a 10 mm long sciatic nerve defect in rats, The 10 mm gap was left empty after resection in the blank group. In the other three groups, the sciatic nerve defect was, respectively, bridged with plain collagen tube (CT group), a collagen tube filled with BMSC suspension (C+ CT group), or reversed transected nerve segment (AG group). \u0000 \u0000 \u0000Results \u0000Histological and functional assessments showed that the developed TENG facilitated nerve regeneration (SFI: CT, -92.490±1.836; C+ CT, -70.010±7.805; AG, -70.130±8.744; F= 17.850, P 0.05) and better outcomes than the plain collagen tube (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000These results manifested that the bilayer collagen conduit loaded BMSCs could be applied as a new biodegradable artificial nerve guide for nerve tissue engineering. \u0000 \u0000 \u0000Key words: \u0000Tissue engineer; Nerve graft; Bone marrow mesenchymal stem cells; Bilayer collagen conduit scaffold; Sciatic nerve injury; Peripheral nerve regeneration","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"105-108"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46777948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.054
C. Luo, Zeyuan Yu, Gang Chen, Long Li, Ruiying Luo, Z. Jiao
{"title":"Expression of ubiquitin-conjugating enzyme E2T in gastric cancer and para-carcinoma tissues","authors":"C. Luo, Zeyuan Yu, Gang Chen, Long Li, Ruiying Luo, Z. Jiao","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.054","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.054","url":null,"abstract":"","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"179-180"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45061753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.005
Chuan-jiang Liu, Peng Xia, Pan Liu, Wen-ping Zhou, Q. Fu, T. Qin, Hongwei Zhang
Objective To observe the expression of microRNA (miRNA, miR)-15b in pancreatic cancer (PC) cells and its effect on proliferation, migration and apoptosis of pancreatic cancer cell lines. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect miR-15b expression in four pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) and human normal pancreatic ductal epithelial cell line (HPDE6c7). CFPAC-1 and PANC-1 cells stably transfected with miR-15b short hairpin RNA (shRNA) were used as experimental group, and CFPAC-1 and PANC-1 cells which normally expressed miR-15b were transfected with negative control (NC) as control group. The proliferation of CFPAC-1 and PANC-1 cells was measured by cell counting kit-8 (CCK-8) proliferation assay after transfection. The migration ability of CFPAC-1 and PANC-1 cells was measured by Transwell migration assay. The apoptosis of CFPAC-1 and PANC-1 cells was examined by flow cytometry. Results The expression levels of miR-15b in 4 pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) were 3.63±1.02, 5.28±0.76, 6.72±1.39 and 4.68±1.56 respectively, which were significantly higher than that (1.08±0.23) in HPDE6c7 cells (t=9.944, 10.326, 12.013, 15.877, P<0.01). The cell proliferation ability of CFPAC-1 cells in miR-15b low expression group at 2, 3 and 4 d (0.51±0.04, 0.94±0.06, 1.25±0.15) was significantly reduced correspondingly as compared with that of CFPAC-1 cells in control group (0.68±0.06, 1.21±0.09, 1.95±0.12) (t=8.256, 6.350, 7.002, P<0.05). The cell proliferation ability of PANC-1 cells in miR-15b low expression group at 2, 3 and 4 d(0.48±0.05, 0.76±0.13, 1.44±0.25) was significantly decreased correspondingly as compared with that in the control group (0.58±0.07, 0.98±0.13, 1.86±0.14) (t=9.988, 10.022, 13.050, P<0.01). The number of CFPAC-1 and PANC-1 cells migrating through the Transwell chamber in the miR-15b low-expression group (38.47±4.69 and 47.83±12.47) was significantly lower than that in the control group (62.39±7.39 and 68.97±8.44) (t=5.798, 8.465, P<0.05). The apoptosis rate of CFPAC-1 and PANC-1 cells in the miR-15b low-expression group [(14.68±3.21)% and (11.57±2.52)%] was significantly higher corresponding than that in the control group [(4.29±1.42)% and (4.73±0.38)%] (t=6.350, 8.666, P<0.05). Conclusion MiR-15b is highly expressed in PC cells. Low expression of miR-15b can decrease the proliferation and migration of PC cells and promote apoptosis of PC cells. Key words: MicroRNA-15b; Proliferation; Migration; Apoptosis
{"title":"Function of microRNA-15b in proliferation, migration and apoptosis of pancreatic cancer cells","authors":"Chuan-jiang Liu, Peng Xia, Pan Liu, Wen-ping Zhou, Q. Fu, T. Qin, Hongwei Zhang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.005","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.005","url":null,"abstract":"Objective \u0000To observe the expression of microRNA (miRNA, miR)-15b in pancreatic cancer (PC) cells and its effect on proliferation, migration and apoptosis of pancreatic cancer cell lines. \u0000 \u0000 \u0000Methods \u0000Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect miR-15b expression in four pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) and human normal pancreatic ductal epithelial cell line (HPDE6c7). CFPAC-1 and PANC-1 cells stably transfected with miR-15b short hairpin RNA (shRNA) were used as experimental group, and CFPAC-1 and PANC-1 cells which normally expressed miR-15b were transfected with negative control (NC) as control group. The proliferation of CFPAC-1 and PANC-1 cells was measured by cell counting kit-8 (CCK-8) proliferation assay after transfection. The migration ability of CFPAC-1 and PANC-1 cells was measured by Transwell migration assay. The apoptosis of CFPAC-1 and PANC-1 cells was examined by flow cytometry. \u0000 \u0000 \u0000Results \u0000The expression levels of miR-15b in 4 pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) were 3.63±1.02, 5.28±0.76, 6.72±1.39 and 4.68±1.56 respectively, which were significantly higher than that (1.08±0.23) in HPDE6c7 cells (t=9.944, 10.326, 12.013, 15.877, P<0.01). The cell proliferation ability of CFPAC-1 cells in miR-15b low expression group at 2, 3 and 4 d (0.51±0.04, 0.94±0.06, 1.25±0.15) was significantly reduced correspondingly as compared with that of CFPAC-1 cells in control group (0.68±0.06, 1.21±0.09, 1.95±0.12) (t=8.256, 6.350, 7.002, P<0.05). The cell proliferation ability of PANC-1 cells in miR-15b low expression group at 2, 3 and 4 d(0.48±0.05, 0.76±0.13, 1.44±0.25) was significantly decreased correspondingly as compared with that in the control group (0.58±0.07, 0.98±0.13, 1.86±0.14) (t=9.988, 10.022, 13.050, P<0.01). The number of CFPAC-1 and PANC-1 cells migrating through the Transwell chamber in the miR-15b low-expression group (38.47±4.69 and 47.83±12.47) was significantly lower than that in the control group (62.39±7.39 and 68.97±8.44) (t=5.798, 8.465, P<0.05). The apoptosis rate of CFPAC-1 and PANC-1 cells in the miR-15b low-expression group [(14.68±3.21)% and (11.57±2.52)%] was significantly higher corresponding than that in the control group [(4.29±1.42)% and (4.73±0.38)%] (t=6.350, 8.666, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000MiR-15b is highly expressed in PC cells. Low expression of miR-15b can decrease the proliferation and migration of PC cells and promote apoptosis of PC cells. \u0000 \u0000 \u0000Key words: \u0000MicroRNA-15b; Proliferation; Migration; Apoptosis","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"19-21"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49337781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.023
Jun Hong, Jingquan Liu, Fangxiao Gong, Lingzhi Jiang, Shijing Mo, Minhua Chen, Xianghong Yang, R. Sun
Objective To investigate the effects of microRNA (miRNA, miR)-224 level changes on the lipopolysaccharide (LPS)-induced injury of pulmonary microvascular endothelium cells (PMVECs) and its mechanism. Methods Primary PMVECs were isolated from clean-level BALB/c mice and cultured in vitro. In order to induce cells injury, 1.0 mg/L LPS was used to treat PMVECs. MiR-224 inhibitor and p21 siRNA were transfected for silencing miR-224 and p21, respectively. The cell viability and apoptosis rate of PMVECs were detected by cell counting kit-8 reagent and flow cytometry, respectively. The changes of miR-224 and p21 were detected by fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting. Dual-luciferase reporter assay was used to determine the target relationship between miR-224 and p21. T-test was used to compare the mean between the two groups and the pairwise comparison of the mean among multiple groups was conducted by LSD test on the basis of one-way analysis of variance. Results After LPS treatment, the relative cell viability decreased to (42.333±7.586)% and apoptosis rate increased to (32.141±2.449)% as compared with non-LPS treated cells. Meanwhile, the level of miR-224 increased to 1.791±0.167 with a decreased p21 mRNA expression (0.527±0.058) (t=8.532, 7.261, 7.113 and 8.467, P<0.01). As compared with simple LPS treated cells, PMVECs transfected with miR-224 inhibitor showed lower cell inhibition and apoptosis rate after LPS treatment (F=62.618 and 32.643, P<0.01). However, p21 knock-down could antagonize the protective effect of miR-224. Conclusion MiR-224 may mediate the LPS-induced injury of PMVECs via targeting p21. MiR-224 could be a therapy target for acute lung injury/acute respiratory distress syndrome after sepsis. Key words: Sepsis; Lipopolysaccharide; Endotheliumcell; Acute lung injury; Acute respiratory distress syndrome; MicroRNA; P21
{"title":"MicroRNA-224 mediates lipopolysaccharide-induced injury of pulmonary microvascular endothelium cells via regulating p21","authors":"Jun Hong, Jingquan Liu, Fangxiao Gong, Lingzhi Jiang, Shijing Mo, Minhua Chen, Xianghong Yang, R. Sun","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.023","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.023","url":null,"abstract":"Objective \u0000To investigate the effects of microRNA (miRNA, miR)-224 level changes on the lipopolysaccharide (LPS)-induced injury of pulmonary microvascular endothelium cells (PMVECs) and its mechanism. \u0000 \u0000 \u0000Methods \u0000Primary PMVECs were isolated from clean-level BALB/c mice and cultured in vitro. In order to induce cells injury, 1.0 mg/L LPS was used to treat PMVECs. MiR-224 inhibitor and p21 siRNA were transfected for silencing miR-224 and p21, respectively. The cell viability and apoptosis rate of PMVECs were detected by cell counting kit-8 reagent and flow cytometry, respectively. The changes of miR-224 and p21 were detected by fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting. Dual-luciferase reporter assay was used to determine the target relationship between miR-224 and p21. T-test was used to compare the mean between the two groups and the pairwise comparison of the mean among multiple groups was conducted by LSD test on the basis of one-way analysis of variance. \u0000 \u0000 \u0000Results \u0000After LPS treatment, the relative cell viability decreased to (42.333±7.586)% and apoptosis rate increased to (32.141±2.449)% as compared with non-LPS treated cells. Meanwhile, the level of miR-224 increased to 1.791±0.167 with a decreased p21 mRNA expression (0.527±0.058) (t=8.532, 7.261, 7.113 and 8.467, P<0.01). As compared with simple LPS treated cells, PMVECs transfected with miR-224 inhibitor showed lower cell inhibition and apoptosis rate after LPS treatment (F=62.618 and 32.643, P<0.01). However, p21 knock-down could antagonize the protective effect of miR-224. \u0000 \u0000 \u0000Conclusion \u0000MiR-224 may mediate the LPS-induced injury of PMVECs via targeting p21. MiR-224 could be a therapy target for acute lung injury/acute respiratory distress syndrome after sepsis. \u0000 \u0000 \u0000Key words: \u0000Sepsis; Lipopolysaccharide; Endotheliumcell; Acute lung injury; Acute respiratory distress syndrome; MicroRNA; P21","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"81-83"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41896948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To establish a stable and reliable mouse atherosclerotic (AS) model, and standardize principal process of AS model identification. Methods The mouse AS model was developed by feeding apolipoprotein E (ApoE) gene knockout mice (ApoE-/-) on high-fat diet (hfd). Sixteen weeks later, the body weight, blood sugar, blood pressure and blood lipid were determined, and aortas were stained with oil red O and hematoxylin to evaluate the AS plaque. Results Cholesterol [(20.09±1.02) mmol/L] and low density lipoprotein cholesterol [(6.84±0.65) mmol/L] of ApoE-/- mice with hfd were significantly increased as compared with that in C57BL/6J normal diet (nd) group [total cholesterol (TC): (2.04±0.07) mmol/L, LDL-C: (0.25±0.01) mmol/L, F=190.543, 82.795, P<0.01]. Oil Red O staining of entire aorta showed that AS lesion size in ApoE-/-+ hfd mice [(22.09±3.49)%] was dramatically increased as compared with that in ApoE-/-+ nd mice [(1.46±0.96)%, F=118.558, P<0.01]. Similar result was obtained from cross-sections of aortic root analysis. Hematoxylin and eosin (HE) staining of cross-sections of aorta root showed typical As plaques. Conclusion The hfd treatment successfully promoted AS development in ApoE-/- mice. The process of gene identification-general situation analysis-blood lipid analysis-morphologic analysis was established to verify AS model. The establishment of mouse AS model provides a valuable tool for the study of AS-related diseases in vivo. Key words: Atherosclerosis; Gene knock out mice; Low density lipoprotein; Cholesterol; Lorta
{"title":"Establishment of mouse atherosclerotic model","authors":"Yiquan Dai, Xiaoxiao Yan, Xiao‐Ru Liu, Yichen Lin, Hongyu Chen","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.050","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.050","url":null,"abstract":"Objective \u0000To establish a stable and reliable mouse atherosclerotic (AS) model, and standardize principal process of AS model identification. \u0000 \u0000 \u0000Methods \u0000The mouse AS model was developed by feeding apolipoprotein E (ApoE) gene knockout mice (ApoE-/-) on high-fat diet (hfd). Sixteen weeks later, the body weight, blood sugar, blood pressure and blood lipid were determined, and aortas were stained with oil red O and hematoxylin to evaluate the AS plaque. \u0000 \u0000 \u0000Results \u0000Cholesterol [(20.09±1.02) mmol/L] and low density lipoprotein cholesterol [(6.84±0.65) mmol/L] of ApoE-/- mice with hfd were significantly increased as compared with that in C57BL/6J normal diet (nd) group [total cholesterol (TC): (2.04±0.07) mmol/L, LDL-C: (0.25±0.01) mmol/L, F=190.543, 82.795, P<0.01]. Oil Red O staining of entire aorta showed that AS lesion size in ApoE-/-+ hfd mice [(22.09±3.49)%] was dramatically increased as compared with that in ApoE-/-+ nd mice [(1.46±0.96)%, F=118.558, P<0.01]. Similar result was obtained from cross-sections of aortic root analysis. Hematoxylin and eosin (HE) staining of cross-sections of aorta root showed typical As plaques. \u0000 \u0000 \u0000Conclusion \u0000The hfd treatment successfully promoted AS development in ApoE-/- mice. The process of gene identification-general situation analysis-blood lipid analysis-morphologic analysis was established to verify AS model. The establishment of mouse AS model provides a valuable tool for the study of AS-related diseases in vivo. \u0000 \u0000 \u0000Key words: \u0000Atherosclerosis; Gene knock out mice; Low density lipoprotein; Cholesterol; Lorta","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"172-175"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44866914","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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