抗病毒RNA干扰(RNA interference,RNAi)机制在哺乳动物中保守存在,宿主通过病毒源siRNA(virus-derived small interfering RNA,vsiRNA)引导Ago2蛋白剪切病毒RNA,从而发挥抗病毒作用.为了建立快速检测宿主抗流感病毒(PR8ΔNS1)和野田村病...抗病毒RNA干扰(RNA interference,RNAi)机制在哺乳动物中保守存在,宿主通过病毒源siRNA(virus-derived small interfering RNA,vsiRNA)引导Ago2蛋白剪切病毒RNA,从而发挥抗病毒作用.为了建立快速检测宿主抗流感病毒(PR8ΔNS1)和野田村病毒(NoVΔB2)特异性siRNA分子的方法,根据病毒特异性siRNAs序列设计茎环引物和扩增引物,筛选出具有高特异性和灵敏度的茎环引物和PCR扩增引物(16768,3280和24~45),成功建立了茎环法RT-qPCR检测哺乳动物病毒源siRNA表达水平的系统.该方法快速简便,准确性高,兼具较高的特异性与灵敏度,可作为小RNA测序替代方案检测siRNA或在测序前对宿主产生的病毒源siRNA进行有效监测与评价.展开更多
Background/Aims:Human invariant natural killer T(iNKT) cells express a TCR Vα 24-Jα Q paired with Vβ 11 and are activated by a surrogate ligand,α-galactosylceramide(α GalCer) .The iNKT cells are involved in the r...Background/Aims:Human invariant natural killer T(iNKT) cells express a TCR Vα 24-Jα Q paired with Vβ 11 and are activated by a surrogate ligand,α-galactosylceramide(α GalCer) .The iNKT cells are involved in the regulation of anti-viral immune responses;however,little is known about their roles in hepatitis C virus(HCV) infection.Methods:We compared the frequency of peripheral iNKT cells and their cytokine producing capacity reactive to α GalCer between chronically HCV-infected patients and healthy subjects.Cytokine production of freshly isolated iNKT cells were analyzed by ELISPOT.Activated iNKT cells were obtained by culture with α GalCer-loaded dendritic cells(DCs) and re-stimulated with them for the measurement of cytokine production.Results:The frequencies of iNKT cells were not different between HCV-infected patients and healthy subjects.The number of fresh IFN-γ-producing iNKT cells reactive to α GalCer was not different between the patients and controls,whereas fresh iNKT cells produced negligible amounts of Th2 cytokines regardless of HCV infection.In response to α GalCer,expanded iNKT cells from the patients secreted IFN-γ comparable in amount to controls,whereas they released significantly more IL-13 than cells from controls.Conclusions:Activated iNKT cells from HCV-infected patients gain more ability to secrete IL-13 than those from healthy subjects.展开更多
慢性活动性肝炎（Chronic active hepatitis简称CAH）是一种严重危害人类健康的疾病,且预后较差。CAH的病因主要有乙型肝炎病毒（HBV）,在我国以HBV相关的CAH约占90％左右,其次非甲非乙型肝炎病毒（NANB）及某些药物,如α—甲基多巴、...慢性活动性肝炎（Chronic active hepatitis简称CAH）是一种严重危害人类健康的疾病,且预后较差。CAH的病因主要有乙型肝炎病毒（HBV）,在我国以HBV相关的CAH约占90％左右,其次非甲非乙型肝炎病毒（NANB）及某些药物,如α—甲基多巴、异烟肼、双醋酚酊和酒精中毒,α<sub>1</sub>—抗胰蛋白酶缺乏、肝豆状核变性等也可引起类似CAH的病变。本文仅就CAH的发病机理的几个因素加以叙述。展开更多
文摘抗病毒RNA干扰(RNA interference,RNAi)机制在哺乳动物中保守存在,宿主通过病毒源siRNA(virus-derived small interfering RNA,vsiRNA)引导Ago2蛋白剪切病毒RNA,从而发挥抗病毒作用.为了建立快速检测宿主抗流感病毒(PR8ΔNS1)和野田村病毒(NoVΔB2)特异性siRNA分子的方法,根据病毒特异性siRNAs序列设计茎环引物和扩增引物,筛选出具有高特异性和灵敏度的茎环引物和PCR扩增引物(16768,3280和24~45),成功建立了茎环法RT-qPCR检测哺乳动物病毒源siRNA表达水平的系统.该方法快速简便,准确性高,兼具较高的特异性与灵敏度,可作为小RNA测序替代方案检测siRNA或在测序前对宿主产生的病毒源siRNA进行有效监测与评价.
文摘Background/Aims:Human invariant natural killer T(iNKT) cells express a TCR Vα 24-Jα Q paired with Vβ 11 and are activated by a surrogate ligand,α-galactosylceramide(α GalCer) .The iNKT cells are involved in the regulation of anti-viral immune responses;however,little is known about their roles in hepatitis C virus(HCV) infection.Methods:We compared the frequency of peripheral iNKT cells and their cytokine producing capacity reactive to α GalCer between chronically HCV-infected patients and healthy subjects.Cytokine production of freshly isolated iNKT cells were analyzed by ELISPOT.Activated iNKT cells were obtained by culture with α GalCer-loaded dendritic cells(DCs) and re-stimulated with them for the measurement of cytokine production.Results:The frequencies of iNKT cells were not different between HCV-infected patients and healthy subjects.The number of fresh IFN-γ-producing iNKT cells reactive to α GalCer was not different between the patients and controls,whereas fresh iNKT cells produced negligible amounts of Th2 cytokines regardless of HCV infection.In response to α GalCer,expanded iNKT cells from the patients secreted IFN-γ comparable in amount to controls,whereas they released significantly more IL-13 than cells from controls.Conclusions:Activated iNKT cells from HCV-infected patients gain more ability to secrete IL-13 than those from healthy subjects.
文摘慢性活动性肝炎（Chronic active hepatitis简称CAH）是一种严重危害人类健康的疾病,且预后较差。CAH的病因主要有乙型肝炎病毒（HBV）,在我国以HBV相关的CAH约占90％左右,其次非甲非乙型肝炎病毒（NANB）及某些药物,如α—甲基多巴、异烟肼、双醋酚酊和酒精中毒,α<sub>1</sub>—抗胰蛋白酶缺乏、肝豆状核变性等也可引起类似CAH的病变。本文仅就CAH的发病机理的几个因素加以叙述。
