目的研究妊娠期糖尿病(GDM)孕妇胎盘组织中性激素结合球蛋白(SHBG)、胰岛素信号转导蛋白及葡萄糖转运蛋白表达,探讨其在GDM发病过程中的作用。方法收集足月妊娠且非肥胖(BMI<25 kg/m2)GDM孕妇(GDM组)和同期糖代谢正常(正常组)孕妇胎...目的研究妊娠期糖尿病(GDM)孕妇胎盘组织中性激素结合球蛋白(SHBG)、胰岛素信号转导蛋白及葡萄糖转运蛋白表达,探讨其在GDM发病过程中的作用。方法收集足月妊娠且非肥胖(BMI<25 kg/m2)GDM孕妇(GDM组)和同期糖代谢正常(正常组)孕妇胎盘组织各10例,应用Western blotting、实时PCR方法检测2组胎盘组织中SHBG和胰岛素信号转导蛋白(IRS-1、ISR-2、PI3K p85α)和葡萄糖转运蛋白(GLUT-1、GLUT-3、GLUT-4)的表达,各指标进行直线回归依存关系分析。结果与正常组比较,GDM组胎盘组织中SHBG m RNA、蛋白表达降低(P<0.05);IRS-1蛋白、IRS-2 m RNA表达降低(P<0.05);PI3K p85α和GLUT-1蛋白及其m RNA表达无统计学差异(P>0.05);GLUT-3蛋白,GLUT-4 m RNA、蛋白表达降低(P<0.05)。直线回归分析结果显示SHBG m RNA与IRS-2 m RNA、PI3K p85αm RNA、GLUT-4 m RNA的表达呈正相关(均P<0.05);IRS-2 m RNA和GLUT-4 m RNA的表达呈正相关(P<0.01);IRS-1 m RNA和PI3K p85αm RNA、GLUT-3 m RNA的表达呈负相关(P<0.05);IRS-2m RNA和GLUT-1 m RNA的表达正相关(P<0.01)。结论 GDM孕妇胎盘中胰岛素信号转导和葡萄糖转运蛋白存在异常,SHBG可能参与胰岛素信号通路的调节,GDM时胎盘滋养细胞合成和分泌SHBG减少,引起胰岛素信号转导通路相关蛋白表达降低,从而导致胰岛素抵抗及GDM的发生。展开更多
Burkholderia mallei is the etiologic agent of glanders in solipeds and humans. Lipopolysaccharide (LPS) is a major component of cell envelop of this pathogen. O-antigen, the most external component of LPS, is a virule...Burkholderia mallei is the etiologic agent of glanders in solipeds and humans. Lipopolysaccharide (LPS) is a major component of cell envelop of this pathogen. O-antigen, the most external component of LPS, is a virulence factor and a protective antigen in many pathogenic bacteria. Two putative proteins named Wzm (integral membrane protein) and Wzt (hydrophilic ATP-binding protein) are believed to make up an ABC-2 transporter of B. mallei that facilitates transport of components of O-antigen from cytosol to outer-membrane. We studied the importance of wzt (encoding Wzt) to growth, LPS O-antigen profile, and pathogenicity of B. mallei. A wzt mutant strain was generated by deleting a portion of the wzt in B. mallei wild type strain ATCC 23344 by gene replacement. Compared to the wild type strain, the wzt mutant displayed slower growth in vitro and less lethality in CD1 mice when inoculated intraperitoneally. The 50% lethal doses (LD50) of the wild type and the wzt mutant strains were 5.9 × 105 and 9.1 × 105 cfu, respectively. CD1 mice inoculated with a non-lethal dose of the wzt mutant produced specific serum immunoglobulins IgG1 and IgG2a and were partially protected against challenge with 11.2 times LD50 of the wild type strain. These findings suggest that the wzt is required for optimal in vitro growth and pathogenesis of B. mallei, and a wzt mutant protects CD1 mice against glanders.展开更多
AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation mark...AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation marker of BCRP expression.METHODS MTT assay was used to filtrate BCRP-mediatedresistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated withdifferent concentration anticancer agents.High performance liquid chromatography(HPLC) was applied tomeasure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest33258,staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs.RESULTSThere were shown increasing durg-resistance to 5-fluorouracil,methotrexate,doxirubicin,pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRPcells(P<0.05,n=3),but shown sensitive to paclitaxel,cisplatin,vincristine,mitomycin and vindesine.Therealso was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil withdifferent expression of BCRP(r=-0.885,P<0.05,n=3).There were shown parallel results ofthat decreasingcellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil byfluorescence dye staining and flow cytometry(P<0.05,n=3),and also shown significate rise of the apoptoticrate of BCRP expression cells after treated with Ko143 (P<0.05,n=3).Every group of cells could be differentextently blocked in phase of G_0/G_1 treated with 5-fluorouracil.CONCLUSION Resistance of 5-fluorouracilcould be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate.Cellularability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.展开更多
文摘目的研究妊娠期糖尿病(GDM)孕妇胎盘组织中性激素结合球蛋白(SHBG)、胰岛素信号转导蛋白及葡萄糖转运蛋白表达,探讨其在GDM发病过程中的作用。方法收集足月妊娠且非肥胖(BMI<25 kg/m2)GDM孕妇(GDM组)和同期糖代谢正常(正常组)孕妇胎盘组织各10例,应用Western blotting、实时PCR方法检测2组胎盘组织中SHBG和胰岛素信号转导蛋白(IRS-1、ISR-2、PI3K p85α)和葡萄糖转运蛋白(GLUT-1、GLUT-3、GLUT-4)的表达,各指标进行直线回归依存关系分析。结果与正常组比较,GDM组胎盘组织中SHBG m RNA、蛋白表达降低(P<0.05);IRS-1蛋白、IRS-2 m RNA表达降低(P<0.05);PI3K p85α和GLUT-1蛋白及其m RNA表达无统计学差异(P>0.05);GLUT-3蛋白,GLUT-4 m RNA、蛋白表达降低(P<0.05)。直线回归分析结果显示SHBG m RNA与IRS-2 m RNA、PI3K p85αm RNA、GLUT-4 m RNA的表达呈正相关(均P<0.05);IRS-2 m RNA和GLUT-4 m RNA的表达呈正相关(P<0.01);IRS-1 m RNA和PI3K p85αm RNA、GLUT-3 m RNA的表达呈负相关(P<0.05);IRS-2m RNA和GLUT-1 m RNA的表达正相关(P<0.01)。结论 GDM孕妇胎盘中胰岛素信号转导和葡萄糖转运蛋白存在异常,SHBG可能参与胰岛素信号通路的调节,GDM时胎盘滋养细胞合成和分泌SHBG减少,引起胰岛素信号转导通路相关蛋白表达降低,从而导致胰岛素抵抗及GDM的发生。
文摘Burkholderia mallei is the etiologic agent of glanders in solipeds and humans. Lipopolysaccharide (LPS) is a major component of cell envelop of this pathogen. O-antigen, the most external component of LPS, is a virulence factor and a protective antigen in many pathogenic bacteria. Two putative proteins named Wzm (integral membrane protein) and Wzt (hydrophilic ATP-binding protein) are believed to make up an ABC-2 transporter of B. mallei that facilitates transport of components of O-antigen from cytosol to outer-membrane. We studied the importance of wzt (encoding Wzt) to growth, LPS O-antigen profile, and pathogenicity of B. mallei. A wzt mutant strain was generated by deleting a portion of the wzt in B. mallei wild type strain ATCC 23344 by gene replacement. Compared to the wild type strain, the wzt mutant displayed slower growth in vitro and less lethality in CD1 mice when inoculated intraperitoneally. The 50% lethal doses (LD50) of the wild type and the wzt mutant strains were 5.9 × 105 and 9.1 × 105 cfu, respectively. CD1 mice inoculated with a non-lethal dose of the wzt mutant produced specific serum immunoglobulins IgG1 and IgG2a and were partially protected against challenge with 11.2 times LD50 of the wild type strain. These findings suggest that the wzt is required for optimal in vitro growth and pathogenesis of B. mallei, and a wzt mutant protects CD1 mice against glanders.
文摘AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation marker of BCRP expression.METHODS MTT assay was used to filtrate BCRP-mediatedresistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated withdifferent concentration anticancer agents.High performance liquid chromatography(HPLC) was applied tomeasure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest33258,staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs.RESULTSThere were shown increasing durg-resistance to 5-fluorouracil,methotrexate,doxirubicin,pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRPcells(P<0.05,n=3),but shown sensitive to paclitaxel,cisplatin,vincristine,mitomycin and vindesine.Therealso was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil withdifferent expression of BCRP(r=-0.885,P<0.05,n=3).There were shown parallel results ofthat decreasingcellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil byfluorescence dye staining and flow cytometry(P<0.05,n=3),and also shown significate rise of the apoptoticrate of BCRP expression cells after treated with Ko143 (P<0.05,n=3).Every group of cells could be differentextently blocked in phase of G_0/G_1 treated with 5-fluorouracil.CONCLUSION Resistance of 5-fluorouracilcould be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate.Cellularability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.
