AIM:To evaluate the effect of efflux pump inhibitors (EPIs) on multidrug resistance of Helicobacter pylori (H. pylori).METHODS: H. pylori strains were isolated and cultured on Brucella agar plates with 10% sheep's...AIM:To evaluate the effect of efflux pump inhibitors (EPIs) on multidrug resistance of Helicobacter pylori (H. pylori).METHODS: H. pylori strains were isolated and cultured on Brucella agar plates with 10% sheep's blood. The multidrug resistant (MDR) H. pylori were obtained with the inducer chloramphenicol by repeated doubling of the concentration until no colony was seen, then the susceptibilities of the MDR strains and their parents to 9 antibiotics were assessed with agar dilution tests. The present study included periods before and after the advent of the EPIs, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), reserpine and pantoprazole), and the minimum inhibitory concentrations (MICs) were determined accordingly. In the same way, the effects of 5 proton pump inhibitors (PPIs), used in treatment of H. pylori infection, on MICs of antibiotics were evaluated.RESULTS: Four strains of MDR H. pylori were induced successfully, and the antibiotic susceptibilities of MDR strains were partly restored by CCCP and pantoprazole, but there was little effect of reserpine. Rabeprazole was the most effective of the 5 PPIs which could decrease the MICs of antibiotics for MDR H. pylori significantly.CONCLUSION: In vitro, some EPIs can strengthen the activities of different antibiotics which are the putative substrates of the efflux pump system in H. pylori.展开更多
建立了免疫磁珠分离(Immunomagnetic separation,IMS)联合实时荧光PCR(real time PCR)快速、灵敏地检测虾中沙门氏菌的方法。用纳米磁珠与抗沙门氏菌多克隆抗体制备免疫磁珠,优化反应条件,建立IMS方法。同时针对沙门氏菌ttr基因合成探...建立了免疫磁珠分离(Immunomagnetic separation,IMS)联合实时荧光PCR(real time PCR)快速、灵敏地检测虾中沙门氏菌的方法。用纳米磁珠与抗沙门氏菌多克隆抗体制备免疫磁珠,优化反应条件,建立IMS方法。同时针对沙门氏菌ttr基因合成探针和引物,构建real time PCR体系,选4株代表性的沙门氏菌检测其特异性和灵敏度。结果显示,免疫磁珠的最适添加量为100μL,免疫磁珠与样品菌液的最佳反应时间为30 min。建立的免疫磁珠分离-实时荧光PCR(IMS-real time PCR)方法特异性高,对于虾中沙门氏菌的检测限为鼠伤寒沙门氏菌5×10~1 CFU/25 g,猪霍乱沙门氏菌5×10~2 CFU/25 g,肠炎沙门氏菌和甲型副伤寒沙门氏菌1×10~2CFU/25 g,检测全程可在6 h内完成。对40份实际样品,IMS-real time PCR方法与国标法检测结果完全一致。建立的IMS-Real time PCR方法用时短、灵敏度高,为水产品中沙门氏菌的快速检测提供了技术支撑,对沙门氏菌引起的食源性疾病的预防和控制具有现实意义。展开更多
目的:探讨一氧化氮(NO)对肠上皮细胞表达紧密连接蛋白Occludin的影响,以研究NO对肠黏膜屏障的作用机制.方法:将NO的供体Sin1与肠上皮细胞株Caco-2共培养24 h,采用MTT方法观察NO对肠上皮细胞的作用,并分别提取细胞蛋白和总RNA,采用免疫...目的:探讨一氧化氮(NO)对肠上皮细胞表达紧密连接蛋白Occludin的影响,以研究NO对肠黏膜屏障的作用机制.方法:将NO的供体Sin1与肠上皮细胞株Caco-2共培养24 h,采用MTT方法观察NO对肠上皮细胞的作用,并分别提取细胞蛋白和总RNA,采用免疫蛋白印迹(Westem blot)蛋白半定量方法和实时定量聚合酶链式反应(RO-PCR)方法检测不同NO浓度对Caco-2细胞表达紧密连接蛋白Occludin蛋白和mRNA表达的影响.结果:随着Sin1浓度升高(125,250,500和1000μmol/L)NO对细胞的杀伤作用产生并逐渐增大,Occludin蛋白表达量和mRNA的相对表达量与无Sin1刺激时蛋白及mRNA的表达量相比明显降低(蛋白:375±0.5,374±0.8,363±0.3.363±0.7 vs 398±0.7;mRNA:0.689±0.01,0.578±0.09,0.554±0.03,0.619±0.04 vs 1,均P<0.01).结论:NO可直接损伤肠上皮细胞,同时以剂量依赖形式在蛋白和分子水平影响紧密连接蛋白Occludin的表达.展开更多
AIM:To observe the changes in interstitial cells of Cajal(ICC) in rats with experimental severe acute pancreatitis(SAP).METHODS:A total of twenty-four SD rats were randomly divided into two groups(n = 12),namely the s...AIM:To observe the changes in interstitial cells of Cajal(ICC) in rats with experimental severe acute pancreatitis(SAP).METHODS:A total of twenty-four SD rats were randomly divided into two groups(n = 12),namely the sham(S) group and the SAP group;the SAP rat model was established by retrograde injection of 5% sodium taurocholate(1.0 mL/kg) into the pancreatic duct.Twenty-four hours later intestinal motility was assessed by testing small intestinal propulsion rate,and then the rats were sacrificed.The pancreas and jejunum were resected and underwent routine pathologic examination.Immunohistochemical staining was used to detect c-kit-positive cells in the jejunum.Expression of c-kit mRNA was detected by real-time polymerase chain reaction,and the expression of c-kit protein was evaluated by Western blotting.Ultrastructure of ICC was evaluated by transmission electron microscopy.RESULTS:There was bleeding,necrosis and a largeamount of inflammatory cell infiltration in pancreatic tissue in the SAP group,while in jejunal tissue we observed a markedly denuded mucosal layer,loss of villous tissue and a slightly dilated muscular layer.The small intestinal propulsion rate was 68.66% ± 2.66% in the S group and 41.55% ± 3.85% in the SAP group.Compared with the S group,the rate of the SAP group decreased sharply.The density of c-kit-positive cells in the SAP group was significantly lower than in the S group;the respective mean densities were 88.47 ± 10.49 in the S group and 56.11 ± 7.09 in the SAP group.The levels of c-kit protein and mRNA were 0.36 ± 0.04 and 1.29 ± 0.91 in the SAP group,respectively,which were significantly lower than those in the S group(0.53 ± 0.06,0.64 ± 0.33,respectively).In the SAP group,ICC profiles showed the same change tendency,such as vacuolation of mitochondria,irregular vacuoles and loosened desmosome-like junctions.CONCLUSION:Decreased c-kit-positive cells and ultrastructural changes in ICC resulting from blockade of the c-kit signaling pathway are involved in the intestinal dysmotili展开更多
基金Supported by Henan Distinguished Junior Scholar Grant,No.074100510017
文摘AIM:To evaluate the effect of efflux pump inhibitors (EPIs) on multidrug resistance of Helicobacter pylori (H. pylori).METHODS: H. pylori strains were isolated and cultured on Brucella agar plates with 10% sheep's blood. The multidrug resistant (MDR) H. pylori were obtained with the inducer chloramphenicol by repeated doubling of the concentration until no colony was seen, then the susceptibilities of the MDR strains and their parents to 9 antibiotics were assessed with agar dilution tests. The present study included periods before and after the advent of the EPIs, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), reserpine and pantoprazole), and the minimum inhibitory concentrations (MICs) were determined accordingly. In the same way, the effects of 5 proton pump inhibitors (PPIs), used in treatment of H. pylori infection, on MICs of antibiotics were evaluated.RESULTS: Four strains of MDR H. pylori were induced successfully, and the antibiotic susceptibilities of MDR strains were partly restored by CCCP and pantoprazole, but there was little effect of reserpine. Rabeprazole was the most effective of the 5 PPIs which could decrease the MICs of antibiotics for MDR H. pylori significantly.CONCLUSION: In vitro, some EPIs can strengthen the activities of different antibiotics which are the putative substrates of the efflux pump system in H. pylori.
文摘建立了免疫磁珠分离(Immunomagnetic separation,IMS)联合实时荧光PCR(real time PCR)快速、灵敏地检测虾中沙门氏菌的方法。用纳米磁珠与抗沙门氏菌多克隆抗体制备免疫磁珠,优化反应条件,建立IMS方法。同时针对沙门氏菌ttr基因合成探针和引物,构建real time PCR体系,选4株代表性的沙门氏菌检测其特异性和灵敏度。结果显示,免疫磁珠的最适添加量为100μL,免疫磁珠与样品菌液的最佳反应时间为30 min。建立的免疫磁珠分离-实时荧光PCR(IMS-real time PCR)方法特异性高,对于虾中沙门氏菌的检测限为鼠伤寒沙门氏菌5×10~1 CFU/25 g,猪霍乱沙门氏菌5×10~2 CFU/25 g,肠炎沙门氏菌和甲型副伤寒沙门氏菌1×10~2CFU/25 g,检测全程可在6 h内完成。对40份实际样品,IMS-real time PCR方法与国标法检测结果完全一致。建立的IMS-Real time PCR方法用时短、灵敏度高,为水产品中沙门氏菌的快速检测提供了技术支撑,对沙门氏菌引起的食源性疾病的预防和控制具有现实意义。
文摘目的:探讨一氧化氮(NO)对肠上皮细胞表达紧密连接蛋白Occludin的影响,以研究NO对肠黏膜屏障的作用机制.方法:将NO的供体Sin1与肠上皮细胞株Caco-2共培养24 h,采用MTT方法观察NO对肠上皮细胞的作用,并分别提取细胞蛋白和总RNA,采用免疫蛋白印迹(Westem blot)蛋白半定量方法和实时定量聚合酶链式反应(RO-PCR)方法检测不同NO浓度对Caco-2细胞表达紧密连接蛋白Occludin蛋白和mRNA表达的影响.结果:随着Sin1浓度升高(125,250,500和1000μmol/L)NO对细胞的杀伤作用产生并逐渐增大,Occludin蛋白表达量和mRNA的相对表达量与无Sin1刺激时蛋白及mRNA的表达量相比明显降低(蛋白:375±0.5,374±0.8,363±0.3.363±0.7 vs 398±0.7;mRNA:0.689±0.01,0.578±0.09,0.554±0.03,0.619±0.04 vs 1,均P<0.01).结论:NO可直接损伤肠上皮细胞,同时以剂量依赖形式在蛋白和分子水平影响紧密连接蛋白Occludin的表达.
文摘AIM:To observe the changes in interstitial cells of Cajal(ICC) in rats with experimental severe acute pancreatitis(SAP).METHODS:A total of twenty-four SD rats were randomly divided into two groups(n = 12),namely the sham(S) group and the SAP group;the SAP rat model was established by retrograde injection of 5% sodium taurocholate(1.0 mL/kg) into the pancreatic duct.Twenty-four hours later intestinal motility was assessed by testing small intestinal propulsion rate,and then the rats were sacrificed.The pancreas and jejunum were resected and underwent routine pathologic examination.Immunohistochemical staining was used to detect c-kit-positive cells in the jejunum.Expression of c-kit mRNA was detected by real-time polymerase chain reaction,and the expression of c-kit protein was evaluated by Western blotting.Ultrastructure of ICC was evaluated by transmission electron microscopy.RESULTS:There was bleeding,necrosis and a largeamount of inflammatory cell infiltration in pancreatic tissue in the SAP group,while in jejunal tissue we observed a markedly denuded mucosal layer,loss of villous tissue and a slightly dilated muscular layer.The small intestinal propulsion rate was 68.66% ± 2.66% in the S group and 41.55% ± 3.85% in the SAP group.Compared with the S group,the rate of the SAP group decreased sharply.The density of c-kit-positive cells in the SAP group was significantly lower than in the S group;the respective mean densities were 88.47 ± 10.49 in the S group and 56.11 ± 7.09 in the SAP group.The levels of c-kit protein and mRNA were 0.36 ± 0.04 and 1.29 ± 0.91 in the SAP group,respectively,which were significantly lower than those in the S group(0.53 ± 0.06,0.64 ± 0.33,respectively).In the SAP group,ICC profiles showed the same change tendency,such as vacuolation of mitochondria,irregular vacuoles and loosened desmosome-like junctions.CONCLUSION:Decreased c-kit-positive cells and ultrastructural changes in ICC resulting from blockade of the c-kit signaling pathway are involved in the intestinal dysmotili
