目的:构建乙型肝炎病毒(hepatitis B virus,HBV)-X蛋白(hepatitis B virus x protein,HBx)和增强型绿色荧光蛋白(enhanced green fluorescentprotein,EGFP)真核融合蛋白表达载体(pHBx-EGFP),转染肝癌Bel7402细胞,观察其表达和定位,为进...目的:构建乙型肝炎病毒(hepatitis B virus,HBV)-X蛋白(hepatitis B virus x protein,HBx)和增强型绿色荧光蛋白(enhanced green fluorescentprotein,EGFP)真核融合蛋白表达载体(pHBx-EGFP),转染肝癌Bel7402细胞,观察其表达和定位,为进一步研究HBx功能提供工具.方法:以pcDNA3.1-HBx为模板,应用PCR和DNA重组技术构建增强型绿色荧光蛋白HBx-EGFP融合蛋白表达载体pHBx-EGFP,经脂质体转染Bel7402细胞,应用倒置荧光显微镜观察融合蛋白表达和定位;同时提取转染pHBx-E G F P24h后的B e l7402细胞总蛋白,应用Western blot技术,鉴定HBx在细胞的表达情况.结果:成功扩增HBx片段插入pEGFP载体,经酶切验证后测序正确;pHBx-EGFP转染Bel7402细胞24h后,荧光显微镜下观察显示HBx-EGFP存在于细胞核周区域,而EGFP则弥散分布于整个细胞;Western blot得到HBx-EGFP、EGFP和HBx目的条带.结论:成功构建pHBx-EGFP真核重组蛋白表达载体,通过检测绿色荧光蛋白标记显示其在Bel7402细胞中表达及定位,并证实pHBx-EGFP载体能在Bel7402细胞正确表达.展开更多
To evaluate the efficacy of interferon (IFN) alpha-2b and lamivudine therapy in children with chronic hepatitis B virus (HBV) infection Method Ten children who developed chronic hepatitis B infection received IFN al...To evaluate the efficacy of interferon (IFN) alpha-2b and lamivudine therapy in children with chronic hepatitis B virus (HBV) infection Method Ten children who developed chronic hepatitis B infection received IFN alpha-2b 10 million international units (IU)/m 2 body surface area, subcutaneously three times a week for six months IFN+lamivudine therapy began to be used in the cases who were unresponsive to IFN treatment Results Among 27 HBsAg (+) subjects in this study, interferon treatment was given to 11 subjects who developed chronic hepatitis One case was excluded from the study due to detection of Herpes type 1 encephalitis At the end of six months of follow-up, complete response was obtained in three (30%) patients and partial response in four (40%) patients, whereas no response was detected in three (30%) patients Fifty percent of the cases experienced serological response, 70% biochemical response, and all (100%) had histological response In three cases started concomitant IFN+lamivudine therapy, HBV-DNA became negative at the second month of treatment Conclusions IFN-alpha and lamivudine can be used for the treatment of chronic hepatitis B infection in children展开更多
文摘目的:构建乙型肝炎病毒(hepatitis B virus,HBV)-X蛋白(hepatitis B virus x protein,HBx)和增强型绿色荧光蛋白(enhanced green fluorescentprotein,EGFP)真核融合蛋白表达载体(pHBx-EGFP),转染肝癌Bel7402细胞,观察其表达和定位,为进一步研究HBx功能提供工具.方法:以pcDNA3.1-HBx为模板,应用PCR和DNA重组技术构建增强型绿色荧光蛋白HBx-EGFP融合蛋白表达载体pHBx-EGFP,经脂质体转染Bel7402细胞,应用倒置荧光显微镜观察融合蛋白表达和定位;同时提取转染pHBx-E G F P24h后的B e l7402细胞总蛋白,应用Western blot技术,鉴定HBx在细胞的表达情况.结果:成功扩增HBx片段插入pEGFP载体,经酶切验证后测序正确;pHBx-EGFP转染Bel7402细胞24h后,荧光显微镜下观察显示HBx-EGFP存在于细胞核周区域,而EGFP则弥散分布于整个细胞;Western blot得到HBx-EGFP、EGFP和HBx目的条带.结论:成功构建pHBx-EGFP真核重组蛋白表达载体,通过检测绿色荧光蛋白标记显示其在Bel7402细胞中表达及定位,并证实pHBx-EGFP载体能在Bel7402细胞正确表达.
文摘To evaluate the efficacy of interferon (IFN) alpha-2b and lamivudine therapy in children with chronic hepatitis B virus (HBV) infection Method Ten children who developed chronic hepatitis B infection received IFN alpha-2b 10 million international units (IU)/m 2 body surface area, subcutaneously three times a week for six months IFN+lamivudine therapy began to be used in the cases who were unresponsive to IFN treatment Results Among 27 HBsAg (+) subjects in this study, interferon treatment was given to 11 subjects who developed chronic hepatitis One case was excluded from the study due to detection of Herpes type 1 encephalitis At the end of six months of follow-up, complete response was obtained in three (30%) patients and partial response in four (40%) patients, whereas no response was detected in three (30%) patients Fifty percent of the cases experienced serological response, 70% biochemical response, and all (100%) had histological response In three cases started concomitant IFN+lamivudine therapy, HBV-DNA became negative at the second month of treatment Conclusions IFN-alpha and lamivudine can be used for the treatment of chronic hepatitis B infection in children
