Objective.To study whether the abilities of hepatitis C virus(HCV)E2 gene immunization to induce humoral and cellular immune responses to E2 protein were affected by hepatitis B virus(HBV)preS gene when they were fuse...Objective.To study whether the abilities of hepatitis C virus(HCV)E2 gene immunization to induce humoral and cellular immune responses to E2 protein were affected by hepatitis B virus(HBV)preS gene when they were fused in DNA-immunized mice.Methods.Mice were immunized with E2,preS-E2(preS gene was upstream of E2 gene),and E2-preS(preS gene was downstream of E2 gene)gene by their eukaryotic expression vectors,respectively.The anti-E2 or anti-preS antibodies were detected using the E2 and preS antigens.The cellular immune response to E2 pro-tein in immunized mice was presented by its survival time after injecting SP2/O myeloma cells expressing HCV E2 protein into the abdominal cavity.Results. Chimeric E2 and preS gene immunization can induce mice to develop anti-preS and anti-E2 antibodies.The number of the mice developing anti-E2 antibody and the antibody titers in preS-E2 gene-injected group were higher than those in E2-preS gene-immunized group.However,the mice injected with E2 gene did not develop the detectable anti-E2 antibodies until 12 weeks after DNA immunization.After the mice was injected with target cells,the average survival time of the mice in the group immunized with E2 gene alone was longer than that of the group injected with E2 gene fused with HBV preS and was significantly longer than that of the control(P< 0.05).Conclusion.HBV preS might be a humoral enhancer that can affect the abilities of HCV E2 protein to in-duce immune responses in DNA-immunized mice.展开更多
目的探究Bcl2相互作用蛋白3(BCL2 Interacting Protein 3,BNIP3)表达变化与正常子宫内膜周期的关系及在子宫内膜异位症(EMs)发病中的意义。方法从GEO数据库获得GSE51981、GSE6364、GSE4888、GSE25628的基因表达谱,通过生物信息学手段比...目的探究Bcl2相互作用蛋白3(BCL2 Interacting Protein 3,BNIP3)表达变化与正常子宫内膜周期的关系及在子宫内膜异位症(EMs)发病中的意义。方法从GEO数据库获得GSE51981、GSE6364、GSE4888、GSE25628的基因表达谱,通过生物信息学手段比较正常子宫内膜周期组增殖期到分泌期基因表达变化,分析基因富集情况以及BNIP3在正常子宫内膜月经周期中的表达变化;比较数据集中正常子宫内膜周期组人群与内异症病人差异趋势表达基因以及各样本中BNIP3的表达,总结其在EMs发病中的意义;取23例Ⅲ⁃ⅣEMs病人正位及异位子宫内膜组织,以及25例因子宫肌瘤行子宫切除术病人的子宫内膜组织作为正常对照,通过实时荧光定量qRT⁃PCR及蛋白质印迹法(Western Blot)检测BNIP3在各组织中的表达,验证BNIP3表达变化同EMs发病的关系。结果GSE4888、GSE6364、GSE51981数据集中,BNIP3在增殖期与分泌期相对表达量分别为100.22±16.54、332.80±32.29,487.68±100.09、1425.83±157.47,8.36±0.69、9.82±0.71,均差异有统计学意义(P<0.0001);GSE25628数据集中,BNIP3在正常子宫内膜组织、EMs病人正位、EMs病人异位子宫内膜组织中的表达量分别为(10.11±0.72)、(9.19±0.56)、(8.52±0.57),三组间对比差异有统计学意义;临床样本中,正常内膜组织、EMs正位内膜组织、EMs异位内膜组织中BNIP3的mRNA表达量分别为1.15±0.57、0.56±0.19、0.36±0.08,三组间对比差异有统计学意义;BNIP3蛋白在正常子宫内膜组织、EMs正位内膜组织、EMs异位内膜组织中的表达水平分别为1.00±0.41、0.58±0.20、0.38±0.12,三组间对比均差异有统计学意义。结论BNIP3在子宫内膜组织中的表达降低与EMs的发生具有相关性。展开更多
Two new C-glucoside flavonoids, namely 8-C-b-D-(2-O-acetyl) glucofuranosyl apigenin and 3-O-acetylvitexin, were isolated from leaves of Crataegus pinnatifida Bge. var. major N. E. Br.. Their structures were elucidated...Two new C-glucoside flavonoids, namely 8-C-b-D-(2-O-acetyl) glucofuranosyl apigenin and 3-O-acetylvitexin, were isolated from leaves of Crataegus pinnatifida Bge. var. major N. E. Br.. Their structures were elucidated by the spectroscopic means and chemical evidence.展开更多
文摘泛素连接酶E4B通过U-box基序可将经泛素活化酶(Ubiquitin-activating enzyme, E1)和泛素结合酶(Ubiquitin-conjugating enzyme, E2)传递的泛素(Ubiquitin, UB)标记至底物蛋白质。探究3个蛋白质:金属蛋白酶M20家族蛋白质2(Peptidase M20 domain-containing protein 2,PM20D2)、多腺苷酸结合蛋白质1(Polyadenylate-binding protein 1,PABPC1)、细胞色素C氧化酶Ⅰ翻译激活剂(Translational activator of cytochrome C oxidase Ⅰ,TACO1)是否可被E4B介导泛素化。从HEK293细胞中提取总RNA,反转录为cDNA,以其为模板调取底物基因并构建重组质粒,利用大肠杆菌表达系统表达并纯化泛素化过程所需的各个相关蛋白质,通过蛋白质体外泛素化实验,对3个蛋白质进行泛素化验证。结果表明3个蛋白质均可以在蛋白质水平被E4B泛素化,证明3个蛋白质都是E4B的底物,为进一步在细胞中研究其泛素化的机理奠定基础。
文摘Objective.To study whether the abilities of hepatitis C virus(HCV)E2 gene immunization to induce humoral and cellular immune responses to E2 protein were affected by hepatitis B virus(HBV)preS gene when they were fused in DNA-immunized mice.Methods.Mice were immunized with E2,preS-E2(preS gene was upstream of E2 gene),and E2-preS(preS gene was downstream of E2 gene)gene by their eukaryotic expression vectors,respectively.The anti-E2 or anti-preS antibodies were detected using the E2 and preS antigens.The cellular immune response to E2 pro-tein in immunized mice was presented by its survival time after injecting SP2/O myeloma cells expressing HCV E2 protein into the abdominal cavity.Results. Chimeric E2 and preS gene immunization can induce mice to develop anti-preS and anti-E2 antibodies.The number of the mice developing anti-E2 antibody and the antibody titers in preS-E2 gene-injected group were higher than those in E2-preS gene-immunized group.However,the mice injected with E2 gene did not develop the detectable anti-E2 antibodies until 12 weeks after DNA immunization.After the mice was injected with target cells,the average survival time of the mice in the group immunized with E2 gene alone was longer than that of the group injected with E2 gene fused with HBV preS and was significantly longer than that of the control(P< 0.05).Conclusion.HBV preS might be a humoral enhancer that can affect the abilities of HCV E2 protein to in-duce immune responses in DNA-immunized mice.
文摘目的探究Bcl2相互作用蛋白3(BCL2 Interacting Protein 3,BNIP3)表达变化与正常子宫内膜周期的关系及在子宫内膜异位症(EMs)发病中的意义。方法从GEO数据库获得GSE51981、GSE6364、GSE4888、GSE25628的基因表达谱,通过生物信息学手段比较正常子宫内膜周期组增殖期到分泌期基因表达变化,分析基因富集情况以及BNIP3在正常子宫内膜月经周期中的表达变化;比较数据集中正常子宫内膜周期组人群与内异症病人差异趋势表达基因以及各样本中BNIP3的表达,总结其在EMs发病中的意义;取23例Ⅲ⁃ⅣEMs病人正位及异位子宫内膜组织,以及25例因子宫肌瘤行子宫切除术病人的子宫内膜组织作为正常对照,通过实时荧光定量qRT⁃PCR及蛋白质印迹法(Western Blot)检测BNIP3在各组织中的表达,验证BNIP3表达变化同EMs发病的关系。结果GSE4888、GSE6364、GSE51981数据集中,BNIP3在增殖期与分泌期相对表达量分别为100.22±16.54、332.80±32.29,487.68±100.09、1425.83±157.47,8.36±0.69、9.82±0.71,均差异有统计学意义(P<0.0001);GSE25628数据集中,BNIP3在正常子宫内膜组织、EMs病人正位、EMs病人异位子宫内膜组织中的表达量分别为(10.11±0.72)、(9.19±0.56)、(8.52±0.57),三组间对比差异有统计学意义;临床样本中,正常内膜组织、EMs正位内膜组织、EMs异位内膜组织中BNIP3的mRNA表达量分别为1.15±0.57、0.56±0.19、0.36±0.08,三组间对比差异有统计学意义;BNIP3蛋白在正常子宫内膜组织、EMs正位内膜组织、EMs异位内膜组织中的表达水平分别为1.00±0.41、0.58±0.20、0.38±0.12,三组间对比均差异有统计学意义。结论BNIP3在子宫内膜组织中的表达降低与EMs的发生具有相关性。
文摘Two new C-glucoside flavonoids, namely 8-C-b-D-(2-O-acetyl) glucofuranosyl apigenin and 3-O-acetylvitexin, were isolated from leaves of Crataegus pinnatifida Bge. var. major N. E. Br.. Their structures were elucidated by the spectroscopic means and chemical evidence.