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| 1 | RNA interference:Antiviral weapon and beyond显示文摘RNA interference (RNAi) is a remarkable type of gene regulation based on sequence-specific targeting and degradation of RNA. The term encompasses related pathways found in a broad range of eukaryotic organisms, including fungi, plants, and animals. RNA interference is part of a sophisticated network of interconnected pathways for cellular defense, RNA surveillance, and development and it may become a powerful tool to manipulate gene expression experimentally. RNAi technology is currently being evaluated not only as an extremely powerful instrument for functional genomic analyses, but also as a potentially useful method to develop specific dsRNA based gene-silencing therapeutics.Several laboratories have been interested in using RNAi to control viral infection and many reports in Nature and in Cell show that short interfering (si) RNAs can inhibit infection by HIV-1, polio and hepatitis C viruses in a sequence-specific manner. RNA-based strategies for gene inhibition in mammalian cells have recently been described, which offer the promise of antiviral therapy. | Quan-Chu Wang Qing-He Nie Zhi-Hua Feng, The Center of Diagnosis and Treatment for Infectious Diseases of Chinese PLA, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi Province, China | 2003 | World Journal of Gastroenterology2003，9，8： | 14 |
| 2 | HCV RNA在外周血单个核细胞中的复制与丙肝慢性化及复发的关系显示文摘通过对外周血单个核细胞(PBMC)中正链及负链HCV RNA的检测来探讨HCV在PBMC中的复制与丙肝慢性化及复发的关系。应用巢式RT-PCR对71例慢性丙型肝炎患者的血清和PBMC进行正负链HCV RNA的检测,其中15例干扰素治疗。并对10例正常人的血清和PBMC进行了HCV RNA检测。71例慢性丙型肝炎患者血清中正链HCV RNA阳性率73.2%,负链HCV RNA阳性率为0,PBMC中正链HCV RNA阳性率为60.5%,负链HCVRNA阳性率为38.0%。15例干扰素治疗患者在治疗后血清正链HCV RNA转阴率为73.3%,PBMC中正链HCVRNA转阴率为45.5%,负链转阴率为71.4%。正常对照组血清及PBMC中HCV RNA阳性率为0。这说明HCV RNA在PBMC中的复制与丙肝慢性化及复发存在一定的关系。干扰素治疗丙肝具有一定疗效,但不能彻底清除病毒。 | 张秀华 | 2006 | 临床肝胆病杂志2006，22，1： | 7 |
| 3 | 丙型肝炎患者抗病毒治疗后外周血单个核细胞内病毒 RNA 的检测及其与疗效的关系显示文摘目的:观察慢性丙型肝炎患者在聚乙二醇干扰素α-2a (Peg IFNα-2a)联合利巴韦林(RBV)抗病毒治疗前、后血清及 PBMC 内 HCV RNA 的变化,初步探讨 PBMC 内 HCV RNA 检测的临床意义。方法2013年6月至2014年12月在广州市第八人民医院感染科就诊的慢性丙型肝炎患者20例,在治疗前、后的不同时间点(0、2、4、12、24、36、48周)采集患者外周血,分离血浆和 PBMC;采用精确荧光定量方法(Cobas TaqMan 实时 PCR)检测丙型肝炎患者血浆 HCV RNA;采用实时-PCR、套式PCR 两种方法分别检测 PBMC 内 HCV RNA。计数资料采用χ^2检验。结果 Cobas 精确定量检测各时间点血浆中 HCV RNA 分别为(5.92±0.86)、(2.77±1.32)、(1.30±1.72)、(0.15±0.47)、0、0和(0.06±0.26)lgIU/mL;各时间点差异有统计学意义(F =148.06,P <0.01)。12周时有18例患者血浆中 HCV RNA 完全清除。套式 PCR 较实时 PCR 阳性率高(均 P <0.01);20例患者血浆病毒载量及PBMC 中 HCV RNA 检测结果发现,PBMC 中 HCV RNA 清除速率较血浆明显延迟,其中2例 PBMC中 HCV RNA 持续性存在并在治疗结束24周后复发。结论慢性丙型肝炎患者 PBMC 中可以检测到HCV RNA,且套式 PCR 检测较实时 PCR 检测阳性率高;抗病毒治疗对 PBMC 内、外的 HCV 均有效,但 PBMC 中 HCV RNA 清除速率较血浆明显延迟;干扰素抗病毒过程中 PBMC 内 HCV 清除缓慢,预示存在治疗后复发的风险。 | 聂静敏 胡凤玉 许敏 陈伟烈 何浩岚 李凌华 蔡卫平 唐小平 | 2016 | 中华传染病杂志2016，34，3： | 6 |
| 4 | Hepatitis C virus lymphotropism and peculiar immunological phenotype:Effects on natural history and antiviral therapy显示文摘Hepatitis C virus(HCV) has been recognized to be both a hepato-and lymphotropic virus.HCV lymphotropism represents an essential lap in the pathogenesis of virusrelated autoimmune and lymphoproliferative disorders,ranging from clonal expansion of B-cells with organ-and non-organ-specific autoantibody production up to overt non-Hodgkin's lymphoma along a continuous step-by-step model of B-cell lymphomagenesis,where the intermediated mixed cryoglobulinemia could be considered as a stage of suppressible antigen-driven lymphoproliferation.HCV infection of lymphoid cells could set up privileged reservoirs able to interfere with the host viral clearance eff iciency and may be implicated in viral recurrence after apparently successful antiviral therapy.The HCV long-lasting extrahepatic replicative state generates an abnormal systemic immunological response,easily detectable by searching simple laboratory and clinical parameters,mainly represented by vasculitis-like skin features and hypocomplementemia.The presence or absence of this hypersensitivity pattern seems to correlate with the antiviral response and could be identif ied as a novel immunological cofactor.Further research is required to fully verify the real impact on therapeutic choice/regimen. | Paolo Conca Giovanni Tarantino | 2009 | World Journal of Gastroenterology2009，15，19： | 5 |
| 5 | Clearance of HCV RNA in peripheral blood mononuclear cell as a predictor of response to antiviral therapy in patients with chronic hepatitis C显示文摘BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication. But in interferon treated patients the HCV RNA in serum sometimes could not show the virus in cells. Such factors as virus genotype, HCV RNA contents in serum, HCV specific cellular immunities after treatment were reported to predict the response to interferon therapy. In most patients, HCV RNA could detect the virus in peripheral blood mononucle-ar cell. The aim of this study was to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after interferon treatment. METHODS: Sixteen patients with chronic hepatitis C were treated with interferon for 24 weeks, and they all get complete responses at 12 weeks of treatment. At the end of treatment, the HCV RNA in PBMC and serum were detected by RT-PCR, and after stopping treatment, HCV RNA in serum was monitored continually. RESULTS: In 9 patients who were HCV RNA positive in their PBMC at the end of treatment, 8 showed serum HCV RNA positive after 24 weeks and another 1 after 1 year. In 7 patients with negative HCV RNA in their PBMC, only 2 patients relapsed in serum HCV RNA after 1-year follow-up, and others remained viral response after 3.5 years. CONCLUSION: HCV RNA in PBMC at the end of IFN treatment is a predictor of durable response to antiviral therapy in patients with chronic hepatitis C. | Dao-Zhen Xu, Yao Xie and Zheng-Qin Li Beijing Ditan Hospital, Beijing 100011, China | 2005 | Hepatobiliary & Pancreatic Diseases International2005，4，4： | 4 |
| 6 | Flow cytometric detection of hepatitis C virus antigens in infected peripheral blood leukocytes: Binding and entry显示文摘AIM: We designed two synthetic-core-specific peptides core 1 (C1) and core 2 (C2), and an E1-specific peptide (E1). We produced specific polyclonal antibodies againstthese peptides and used the antibodies for detection of HCV antigens on surface and within infected peripheral blood leukocytes.METHODS: Peripheral blood from a healthy individual who tested negative for HCV RNA was incubated with HCV type 4 infected serum for 1 h and 24 h at 37 ℃. Cells were stained by direct and indirect immunofluorescence and measured by flow cytometry.RESULTS: After 1 h of incubation, antibodies against C1,C2, and E1 detected HCV antigens on the surface of 27%,26% and 73% of monocytes respectively, while 10%, 5% and 9% of lymphocytes were positive with anti-C1, anti-C2 and anti-E1 respectively. Only 1-3% of granulocytes showed positive staining with anti-C1, anti-C2 and anti E1 antibodies. After 24 h of incubation, we found no surface staining with anti-C1, anti-C2 or anti-E1. Direct immunostaining using anti-C2 could not detect intracellular HCV antigens, after 1 h of incubation with the virus, while after 24 h of incubation, 28% of infected cells showed positive staining. Only plus strand RNA was detectable intracellularly as early as 1 h after incubation, and remained detectable throughout 48 h post-infection.Interestingly, minus RNA strand could not be detected after 1 h, but became strongly detectable intracellularly after 24 h post-infection.CONCLUSION: Monocytes and lymphocytes are the preferred target cells for HCV infection in peripheral blood leukocytes. Our specific anti-core and anti-E1 antibodies are valuable reagents for demonstration of HCV cell cycle.Also, HCV is capable of infecting and replicating in peripheral blood mononuclear cells as confirmed by detection of minus strand HCV RNA as well as intracellular staining of core HCV antigen. | Mostafa K El-Awady Ashraf A Tabll El-Rashdy M Redwan Samar Youssef Moataza H Omran Fouad Thakeb Maha El-Demellawy | 2005 | World Journal of Gastroenterology2005，11，33： | 4 |
| 7 | 丙型肝炎患者外周血单核细胞内病毒载量与干扰素治疗效果的相关性研究显示文摘目的探讨慢性丙型肝炎患者外周血单核细胞(PBMCs)中病毒载量,观察聚乙二醇干扰素(PEG-IFNa-2 a)抗病毒治疗后PBMCs中HCVRNA的变化及其临床意义。方法采用荧光定量方法检测丙型肝炎患者血清及外周血单核细胞内病毒载量。结果血清中HCVRNA阳性患者其PBMCs中检出率为83.6%,HCV RNA阴性患者检出率为16.3%;治疗结束及停药6个月时丙氨酸氨基转移酶(ALT)复常率、PBMCs中HCVRNA阴转率均与治疗前比较下降明显(P<0.05),PEG-IFNa-2 a对血清及PBMCs中的HCVRNA均有明显的抑制作用,但PBMCs中HCVRNA清除速率较血清明显延迟。结论PBMCs是HCV在肝外复制的重要场所,是评价HCV活动最理想的标志,可作为临床评价病毒复制程度和抗病毒疗效的参考指标。 | 范公忍 韩聚强 熊锦华 任永强 胡学玲 | 2008 | 临床内科杂志2008，25，9： | 3 |
| 8 | 丙型肝炎病毒NS5A基因变异与干扰素疗效的关系显示文摘目的:探讨HCV1b型慢性丙型肝炎患者HCV NS5A基因变异与干扰素(IFN)疗效的关系,NS5A_(2209-2248)片断是否存在干扰素敏感决定区(ISDR)。方法:留取慢性丙型肝炎患者干扰素治疗前血清,应用RT-PCR法扩增NS5A基因片段,用直接测序法进行核苷酸及氨基酸序列测定。结果:11例HCV1b型患者中1例为中间型,其余为野生型。2例表现为完全应答,均为野生型,9例为无应答。两组之间核苷酸及氨基酸序列无显著差异。对1例患者的动态观察发现,干扰素治疗后其核苷酸及氨基酸序列均有所变化,改变了其在基因树中的位置。结论:HCV1b型NS5A基因变异与干扰素疗效无关,NS5A_(2209-2248)区高度保守。未证实存在ISDR。干扰素治疗可引起HCV准种改变。 | 张琳 赵桂珍 石理兰 曹丽 | 2003 | 世界华人消化杂志2003，11，8： | 2 |
| 9 | Expression of hepatitis C virus hypervariable region 1 and its clinical significance显示文摘AIM: To explore the properties of hypervariable region 1(HVR1) in the envelope 2 gene of hepatitis C virus by analyzing the reactivity of HVR1 fusion proteins from different Chinese HCV strains with sera of patients with chronic hepatitis C and by comparing their reactivity between interferon therapy responders and non-responders.METHODS: Gene fragments of HVR1 of four HCV strains (three genotype 1b and one genotype 2a) were amplified from pGEMT-E2 plasmids and sub-cloned into pQE40vectors respectively to construct recombinant expression plasmids which expressed HVR1 fused downstream to DHFR in Escherichia coli strain TG1. The purified DHFRHVR1 proteins were then used to detect the anti-HVR1antibodies in 70 serum samples of patients with chronic hepatitis C.RESULTS: Four DHFR- HVR1 fusion proteins were successfully expressed in E.coli (320-800 ug fusion proteins per 100 ml culture). Each fusion protein (SH1b, BJ1b,SD1b and SD2a) reacted with 72.8 % (51/70), 60 % (42/70), 48.6 % (34/70), and 58.6 % (41/70) of the anti-HCV positive patients' sera respectively by ELISA. 57.1% (4/7) of non responders reacted with all four HVR1 fusion proteins, while only 15.3 % (2/13) of responders reacted with all of them. The O.D. values of sera from IFN therapy responders were significantly higher than those of non responders (P<0.05).CONCLUSION: The selected HVR1 fusion proteins expressed in E. coli can broadly react with HCV-infected patients' sera. The intensity and/or quality of the immune response against HCV may be a critical factor determining the response to interferon treatment. With the evolution of virus strains, anti-HVR1 antibodies can not neutralize all the quasispecies. A polyvalent and high immunogenic vaccine comprising a mixture of several HVR1 sequences that cover the reactivity of most HCV isolates may be useful. | Xin-XinZhang Shen-YigZhang JingLiu Zhi-MengLu YuanWang | 2003 | World Journal of Gastroenterology2003，9，5： | 2 |
| 10 | Monocyte chemotactic protein-1 and soluble adhesion molecules as possible prognostic markers of the efficacy of antiviral treatment in chronic hepatitis C显示文摘AIM: To explain the role of Monocyte chemotactic protein-1 (MCP-1) and soluble adhesion molecules in chronic hepatitis C during the treatment of interferon alpha (IFNα) 2 b and ribavirin (RBV).METHODS: Concentrations of MCP-1, soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1), sPselectin, interleukin (IL) 6, and ILl0 in serum were estimated in the group of 40 patients with chronic hepatitis C treated with IFNalpha2 b and RBV in 0, 16, 32, 48 wk of the therapy.RESULTS: In chronic hepatitis C, before and during the treatment, the serum levels of MCP-1 and sP-selectin in responders were similar to those of healthy subjects. In nonresponders (NR), MCP-1 increased in the course of IFN(α+RBV treatment, differences were statistically significant as compared to responders. MCP-1 correlated statistically with the activity of periportal inflammation (r= 0.35, P<0.05) but not with staging of liver fibrosis, sICAM-1 positively correlated with inflammatory activity and fibrosis in NR. sP-selectin did not correlate with histological findings in the liver. The MCP-1 correlated with the soluble form of sP-selectin concentrations (r= 6, P<0.001) and with IL-10 level in NR (r = 0.4, P<0.05). There was no correlation observed between the concentration of MCP-1 and sICAM-1, IL-6 during the treatment.CONCLUSION: MCP-1 concentration may be a prognostic marker of the efficacy of IFN+RBV therapy in patients with chronic hepatitis C. | AnatolPanasiuk DanutaProkopowicz BozenaPanasiuk | 2004 | World Journal of Gastroenterology2004，10，24： | 1 |
| 11 | IFN治疗慢性丙型病毒性肝炎患者外周血单个核细胞HCV的变化显示文摘目的:应用荧光定量反转录多聚酶链反应技术(RT-PCR)及免疫组化技术研究慢性丙型病毒性肝炎患者外周血单个核细胞(PBMC)丙型肝炎病毒感染并评价干扰素对外周血单个核细胞中HCV的作用. 方法:20例慢性丙型病毒性肝炎患者血浆、PBMC中的HCV RNA含量应用荧光定量RT-PCR检测,应用免疫组化技术检测HCV NS3在PBMC中的表达;荧光定量RT- PCR法检测8例血浆及PBMC均阳性患者接受α-2b干扰素治疗;对照组6例血浆及PBMC荧光定量RT-PCR法检测均阳性,未用干扰素治疗.比较治疗前后用药组与对照组血浆及PBMC中HCV RNA载量变化.应用双侧Wilcoxon 秩和检验(wilcoxon 2-sample test),确切概率法(fisher’s exact test)进行数据分析. 结果:20例中荧光定量RT-PCR检出血浆15例、PBMC9 例阳性;免疫组化法检出7例PBMC阳性.应用确切概率法分析PBMC感染HCV RNA与血浆中病毒水平无相关性(P=0.319).8例患者接受干扰素治疗后其血浆及PBMC 中HCV RNA载量均下降,与治疗前相比有显著性差异(aP=0.0017,bP=0.0 059);与对照组相比治疗后差异有显著性(cP=0.0 042,dp=0.0 155). 结论:PBMC是丙型肝炎病毒肝外复制并表达的场所. PBMC感染HCV RNA病毒载量与血浆病毒水平无关.干扰素对PBMC感染的HCV RNA有清除作用. | 付丽娟 吕淑兰 程险峰 王晓燕 | 2004 | 世界华人消化杂志2004，12，3： | 1 |
| 12 | 丙型肝炎患者外周血单个核细胞内HCV RNA的定量测定显示文摘目的:明确丙型肝炎患者外周血单个核细胞内是否可检测到HCV RNA,以及外周血清与PBMCs中HCV RNA载量的相关性。方法:通过SYBR Green Realtime PCR对丙型肝炎患者外周血单个核细胞内及血清中HCV RNA进行定量检测。结果:40例抗HCV阳性的患者中35例血清HCVRNA阳性,5例血清HCVRNA阴性,35例血清HCVRNA阳性的患者中30例PBMCs中可测定到HCVRNA,而5例血清HCVRNA阴性的患者中2例PBMCs中检测到HCVRNA。统计分析示患者外周血清与PBMCs中HCVRNA载量无相关性(P>0.05)。结论:HCV患者PBMCs中可测定到HCV RNA,外周血清与PB-MCs中HCV RNA载量无相关性。 | 郑嵘炅 邢英 李刚 张跃新 | 2011 | 新疆医学2011，41，10： | 0 |
| 13 | 洗涤方法和冻存温度对外周血单个核细胞内HCV RNA含量的影响显示文摘目的分析洗涤方法和冻存温度对外周血单个核细胞(PBMC)内HCVRNA含量的影响。方法收集同一天内17例血清HCVRNA阳性慢性丙型肝炎患者PBMC,随机分成A组(6例)和B组(11例),A组把细胞分成2份后分别采用常规法及改良法洗涤细胞5次,并留取每次洗涤液及细胞;B组采用改良法洗涤细胞后把细胞分成2份,分别作为新鲜组和-20℃冻存组,然后采用离心柱型RNA提取法和实时荧光定量聚合酶链反应(FQPCR)检测洗涤液和细胞中HCVRNA含量。结果(1)洗涤完3次时,常规法有1份洗涤液HCV为阳性,改良法均为阴性;与第1次洗涤液相比,改良法再洗涤1次的病毒含量下降幅度明显高于常规法;(2)新鲜组与冻存组HCVRNA含量不存在明显差异。结论洗涤PBMC以采用改良法洗涤3次为宜;-20℃简易冻存6周的PBMC标本仍可用于FQPCR检测。 | 钟华平 赵伟 沈玲 薛蓉 吴引伟 殷艳天 | 2008 | 江苏医药2008，34，6： | 0 |
| 14 | α2b-IFN诱导CD25表达和对PBMC内HCV RNA的阴转作用显示文摘目的探讨α2b-IFN对PBMC内CD25的诱导及对HCV-RNA、抗-HCV转阴效果。方法用BSA法检测慢性丙肝患者治疗前后CD25表达水平。以α2b-IFN(3MU/d)治疗3个月为一疗程,共2个疗程,于治疗前后分别检测患者PBMC内HCV-RNA和血清HCV-RNA、抗-HCV。结果慢性丙肝患者α2b-IFN治疗后静息期、诱导期CD25表达水平分别为(3.44±0.77)%、(33.62±3.95)%,PBMC、血清内HCV-RNA和抗-HCV转阴率分别为42.31%(11/26)、57.69%(15/26)和65.38%(17/26),与对照相比差异有显著性(P〈0.01~P〈0.05)。结论α2b—IFN可诱导CD25表达,对PBMC内HCV-RNA具有肯定的治疗作用,其疗效优于常规治疗。 | 王健 王瑜 江水清 项桂菊 | 2006 | 中国微生态学杂志2006，18，1： | 0 |
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