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浙江大学学报（医学版） 2011年第6期40卷 588-592,652页

作者：李苏娟陈智朱海红浙江大学医学院附属第一医院传染病诊治国家重点实验室浙江杭州310003

目的:探讨miR-122对α-干扰素(IFN-α)抗丙型肝炎病毒(HCV)效应的影响。方法:给感染HCV的Huh7.5.1予miR-122模拟物(20 nmol/L、100 nmol/L、400 nmol/L)和(或)IFN-α(1 000 IU/ml)处理,采用荧光定量PCR检测HCV RNA水平;给Huh7.5.1细胞感染不同数量HCV(107拷贝,106拷贝,105拷贝)和(或)IFN-α处理,荧光定量PCR检测HCV RNA水平。结果:IFN-α以时间依赖方式抑制HCV复制,在48 h时使HCV RNA下降了83%。miR-122模拟物呈剂量依赖性促进HCV RNA复制(P<0.05)。IFN-α的抗病毒效应与miR-122模拟物水平(20 nmol/L、100nmol/L、400 nmol/L)成反比,与对照组比较有显著性差异(73.3%±3.5%,64.67%±5.5%,56.33%±5.1%vs 84%±4.5%,后两组P<0.05)。IFN-α的抗病毒效应与HCV载量呈反比(105拷贝组vs 107拷贝组,P<0.05)。结论:在细胞感染模型中,miR-122促进HCV的复制,并且,miR-122的表达可部分中和IFN-α的抗HCV效应。

关键词：肝炎病毒属/药物作用干扰素α/治疗应用微RNAs/治疗应用丙型肝炎病毒 miR-122 IFN-α 病毒载量抗病毒效应

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Genetic diversity and response to IFN of the NS3 protease gene from clinical strains of the hepatitis C virus

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ARCHIVES OF VIROLOGY 2002年第7期147卷 1385-1406页

作者： Holland-Staley, CA Kovari, LC Golenberg, EM Pobursky, KJ Mayers, DL Henry Ford Hosp Detroit MI 48202 USA Wayne State Univ Dept Biochem & Mol Biol Detroit MI USA Wayne State Univ Dept Biol Sci Detroit MI 48202 USA

The N-terminal one-third of the hepatitis C virus nonstructural gene 3 (NS3) codes for a serine protease. To investigate natural genetic diversity of this enzyme a nested PCR reaction was developed to obtain NS3 protease sequence data directly from patient strains. This data was used to determine genetic diversity, phylogenetic and evolutionary rates, and selection of variants by interferon therapy. The potential effect of genetic diversity on enzyme structure using molecular modeling was also attempted. Results show significant variability in clinical HCV strains at both the nucleotide (30.2% for la and 25.8% for 1b) and amino acid sequences (11.0% for 1a and 9.9% for 1b). Phylogenic analysis shows two distinct clades with two HCV isolates grouping as a sister clade to 1b. Structural analysis reveals that most mutations lie in the N-terminus of the enzyme. When strains were sorted as to whether or not the patient had received antiviral therapy, no difference was found in the number or locations of mutations in la strains. However, 1b strains demonstrated an overall drop in the number of positions that were mutated. This study demonstrates significant differences among natural strains that may pose a problem for structure based drug development.

关键词：氨基酸序列抗病毒药/治疗应用药物疗法联合基因病毒肝炎病毒属/药物作用肝炎病毒属/酶学肝炎病毒属/遗传学肝炎丙型/药物疗法肝炎丙型/病毒学干扰素α/治疗应用模型分子分子序列数据种系发生逆转录聚合酶链反应利巴韦林/治疗应用序列比对变异(遗传学) 病毒非结构蛋白质类/化学病毒非结构蛋白质类/遗传学成年人老年人女(雌)性人类男(雄)性中年人

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Internal initiation of translation efficiency in different hepatitis C genotypes isolated from interferon treated patients

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ARCHIVES OF VIROLOGY 1999年第2期144卷 215-229页

作者： Sáiz, JC Lopez de Quinto, S Ibarrola, N López-Labrador, FX Sánchez-Tapias, JM Rodés, J Martínez-Salas, E Univ Autonoma Madrid CSIC Ctr Biol Mol Severo Ochoa E-28049 Madrid Spain Univ Barcelona Hosp Clin IDIBAPS Dept MedLiver Unit Barcelona Spain

Initiation of translation of hepatitis C viral RNA occurs internally and it is mediated by a segment of about 330 nucleotides termed Internal Ribosome Entry Site (IRES) located in the 5' end region. While being the most conserved part of the genome, this region also accumulates nucleotide substitutions which are often covariant. In this study we have examined the activity and sequence variation of IRES elements belonging to genotypes 1b, 2a/2c and 3a in patients that responded or not to interferon therapy. The substitutions found in the IRES region analyzed were predicted to maintain the secondary structure of the RNA. Comparison of their efficiency to promote internal initiation of translation in bicistronic constructs supported the conclusion that for both Ib and 3a genotypes, response to interferon therapy and IRES activity are unrelated, although sequence homology was not always found among isolates from patients with different type of response. IRES activity of the studied genotypes varied about 4-fold under the conditions used in our in vivo assays depending on the cell line used for transfection. Such differences were not evidenced in vitro suggesting that the differences observed depend on trans-acting factors present in the transfected cell.

关键词：抗病毒药/治疗应用碱基序列结合部位 COS细胞细胞系氯霉素O-乙酰转移酶/遗传学氯霉素O-乙酰转移酶/代谢 DNA 互补/化学 DNA 互补/遗传学基因表达调控基因型 HeLa细胞肝炎病毒属/药物作用肝炎病毒属/遗传学肝炎病毒属/分离和提纯肝炎丙型/治疗干扰素类/治疗应用萤光素酶类/遗传学萤光素酶类/代谢分子序列数据肽链起始翻译蛋白质生物合成/遗传学 RNA 病毒/遗传学 RNA 病毒/代谢重组融合蛋白质类/遗传学重组融合蛋白质类/代谢逆转录聚合酶链反应核糖体/代谢序列比对序列分析 DNA 序列同源性核酸肿瘤细胞培养的变异(遗传学) 成年人动物女(雌)性人类男(雄)性中年人

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Management of recurrent hepatitis C after liver transplantation: A concise review

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AMERICAN JOURNAL OF GASTROENTEROLOGY 2000年第9期95卷 2164-2170页

作者： Szabo, G Katz, E Bonkovsky, HL Univ Massachusetts Sch Med Dept Med Div Digest Dis & Nutr Worcester MA 01655 USA Univ Massachusetts Mem Hlth Care Dept Med Liver Biliary & Pancreat Ctr Liver Transplantat Program Worcester MA 01655 USA Univ Massachusetts Mem Hlth Care Dept Surg Liver Biliary & Pancreat Ctr Liver Transplantat Program Worcester MA 01655 USA

Recurrent hepatitis C infection and subsequent graft failure are increasingly recognized problems after orthotopic liver transplantation. Although many prospective therapeutic, controlled trials in primary hepatitis C disease have been reported, large-scale studies are yet to be performed in patients with posttransplant recurrent hepatitis C after liver transplantation. In this review, we summarize the current literature on the therapeutic approaches for recurrent hepatitis C and discuss the results of published studies on therapy with ribavirin or interferon (IFN) alone and on combination therapy with IFN plus ribavirin. Further, we discuss results of prophylactic approaches to the problem of recurrent hepatitis C after transplant. Finally, we discuss additional aspects of anti-hepatitis C virus therapy after liver transplantation. (Am J Gastroenterol 2000;95:2164-2170. (C) 2000 by Am. Cell. of Gastroenterology).

关键词：抗病毒药/治疗应用药物疗法联合肝炎病毒属/药物作用肝炎病毒属/遗传学肝炎丙型慢性/药物疗法肝炎丙型慢性/病因学干扰素α/治疗应用预后 RNA 病毒/分析复发/预防和控制利巴韦林/治疗应用人类

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Mutations in interferon sensitivity determining region of hepatitis C virus: Its relation to change in viral load

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AMERICAN JOURNAL OF GASTROENTEROLOGY 2000年第7期95卷 1781-1787页

作者： Terazaw, Y Yoshioka, K Kobayashi, M Watanabe, K Ishigami, M Yano, M Takagi, K Kakumu, S Nagoya Univ Sch Med Dept Internal Med 3 Showa Ku Nagoya Aichi 4668550 Japan Aichi Med Sch Dept Internal Med 1 Aichi Japan

OBJECTIVE: Interferon sensitivity-determining region (ISDR) in nonstructural region 5A of hepatitis C virus (HCV) genotype-lb has been reported to be associated with viral load. Viral load is usually small in the patients with mutant type (four or more amino acid substitutions, compared with HCV-J) and large in those with wild (identical to HCV-J) or intermediate type (from one to three amino acid substitutions). A possible correlation was investigated between mutations in ISDR and alterations of viral load during the course of disease. METHODS: The sequences of ISDR were determined in eight patients with significant changes of viral load and in 11 patients without changes. RESULTS: In two of the eight patients with significant alterations of viral load, ISDR sequences changed significantly. In one patient whose viral load increased after a course of interferon therapy, the number of substitutions, compared with HCV-J, decreased from five to zero or one;the type of ISDR converted from mutant type to wild or intermediate type. In one patient whose viral load decreased significantly after two courses of interferon therapy, the number of substitutions increased from one to six;ISDR changed from intermediate type to mutant type. In the remaining six patients with changes of viral load and in the other 11 patients without changes, the sequences of ISDR did not change significantly. CONCLUSIONS: The mutations in ISDR are one of the viral factors involved in the changes in viral load during the course of disease, although the majority of other factors involved are still unknown. (C) 2000 by Am. Coll. of Gastroenterology).

关键词：氨基酸序列肝炎病毒属/药物作用肝炎病毒属/遗传学肝炎丙型/血液肝炎丙型/病毒学干扰素类/治疗应用微生物敏感性试验突变病毒载量成年人女(雌)性人类男(雄)性中年人

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ISDR pattern and evolution in patients with chronic hepatitis C treated with standard or Peg-IFN plus ribavirin

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ANTIVIRAL THERAPY 2003年第2期8卷 105-110页

作者： Cappiello, G Abbate, I Lo Iacono, O Longo, R Solmone, M Ferraro, D Antonucci, G Di Marco, V Di Stefano, R Croxi, A Ippolito, G Capobianchi, MR Natl Inst Infect Dis L Spallanzani Rome Italy S Pertini Hosp Rome Italy Univ Palermo Palermo Italy

The aim of the study was to characterize the interferon sensitivity determining region (ISDR) mutation pattern and its changes at 4 weeks of treatment in a population of patients infected with hepatitis C virus (HCV) genotype 1b receiving standard or PEG-IFN plus ribavirin (RBV), to find possible early correlates of therapy outcome. Forty-five patients with chronic hepatitis due to HCV 1b were treated by PEG-IFN-alpha 2b (n=23) or IFN-alpha 2b (n=22) plus RBV 1000-1200 mg/day. They were classified 24 weeks after stopping therapy as sustained responders (SR), relapsers (REL) or non-responders (NR). Sixteen patients were SIR, 12 REL and 17 *** mutations were evaluated by direct sequencing at baseline in all and after 4 weeks in patients with detectable viraemia (n=30).

关键词：氨基酸序列抗病毒药/治疗应用队列研究药物疗法联合基因型肝炎病毒属/药物作用肝炎病毒属/遗传学丙型肝炎慢性/药物疗法干扰素α/治疗应用分子序列数据突变聚乙烯二醇类/治疗应用重组蛋白质类利巴韦林/治疗应用序列比对治疗结果病毒非结构蛋白质类/遗传学女(雌)性人类男(雄)性中年人

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Preventing the spread of hepatitis B and C viruses: Where are germicides relevant?

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AMERICAN JOURNAL OF INFECTION CONTROL 2001年第3期29卷 187-197页

作者： Sattar, SA Tetro, J Springthorpe, VS Giulivi, A Univ Ottawa Fac Med Ctr Res Environm Microbiol Ottawa ON K1H 8M5 Canada Hlth Canada Div Bloodborne Pathogens Ottawa ON K1A 0L2 Canada

Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the most prevalent bloodborne pathogens. Infections caused by these organisms can become chronic and may lead to liver cirrhosis and carcinoma. Limited chemotherapy is now available, but only HBV can be prevented through vaccination. Both viruses are enveloped and relatively sensitive to many physical and chemical agents;their ability to survive in the environment may not be as high as often believed. As a result, their spread occurs mainly through direct parenteral or percutaneous exposure to tainted body fluids and tissues. Careful screening of and avoiding contact with such materials remain the most effective means of protection. Nevertheless, the indirect spread of these viruses, although much less common, can occur when objects that are freshly contaminated with tainted blood enter the body or contact damaged skin. Germicidal chemicals are important in the prevention of HBV and HCV spread through shared injection devices, sharps used in personal services (such as tattooing and body piercing), and heat-sensitive medical/dental devices (such as flexible endoscopes) and in the cleanup of blood spills. Microbicides in vaginal gels may also interrupt their transmission. General-purpose environmental disinfection is unlikely to play a significant role in the prevention of the transmission of these viruses. Testing of low-level disinfectants and label claims for such products against HBV and HCV should be discouraged. Both viruses remain difficult to work with in the laboratory, but closely related animal viruses (such as the duck HBV) and the bovine viral diarrhea virus show considerable promise as surrogates for HBV and HCV, respectively. Although progress in the culturing of HBV and HCV is still underway, critical issues on virus survival and inactivation should be addressed with the use of these surrogates.

关键词：消毒药(剂)/药理学抗药性微生物肝炎病毒属/药物作用肝炎乙型/预防和控制肝炎乙型/传播肝炎病毒乙型/药物作用肝炎丙型/预防和控制肝炎丙型/传播微生物敏感性试验动物人类

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α-ketoamides, α-ketoesters and α-diketones as HCVNS3 protease inhibitors

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BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 2000年第8期10卷 711-713页

作者： Han, W Hu, ZL Jiang, XJ Decicco, CP Dupont Merck Pharmaceut Co Expt Stn Dept Chem & Phys Sci Wilmington DE 19880 USA

Peptide-based alpha-ketoamides, alpha-ketoesters and alpha-diketones were designed, synthesized and evaluated against HCV NS3 protease. alpha-Ketoamides have the highest affinity among the three classes, with 8 being the most potent inhibitor with an IC50 Of 340 nM. (C) 2000 DuPont Pharmaceuticals Company. Published by Elsevier Science Ltd. All rights reserved.

关键词：酰胺类/化学氨基酸序列酯类/化学肝炎病毒属/药物作用肝炎病毒属/代谢酮类/化学酮类/药理学丝氨酸蛋白酶抑制剂/化学丝氨酸蛋白酶抑制剂/药理学病毒非结构蛋白质类/药理学

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Follow-up of hepatitis C virus RNA in peripheral blood mononuclear cells during interferon therapy

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ARCHIVES OF VIROLOGY 1999年第2期144卷 355-364页

作者： Cribier, B Uhl, G Schmitt, C Doffoël, M Vetter, D Kirn, A Stoll-Keller, F Univ Hosp INSERM U74 F-67000 Strasbourg France Univ Hosp Serv Hepatogastroenterol Strasbourg France

Mononuclear cells can be infected in vitro by hepatitis C virus and the viral RNA can be detected in mononuclear cells of chronically infected patients. It was suggested that the virus could persist in the mononuclear cells of some patients treated by interferon. The aim of this study was to follow the presence of viral RNA in the plasma and peripheral bleed mononuclear cells of 16 chronically infected patients treated by alpha2b interferon for 1 year. The RNA was detected by reverse transcription followed by nested PCR and quantified using the branched DNA method at regular intervals for at least one year. Before PCR, the mononuclear cells were treated by RNase and trypsin in order to eliminate the viral particles that could be stuck at the cell surface. Six patients were non responders and had persistent plasmatic viral RNA during the treatment. Two patients were good responders and had persistantly negative PCR in both plasma and mononuclear cells. Eight patients had initial negativation of plasmatic hepatitis C virus RNA but showed a relapse characterized by positive plasmatic PCR. Positive PCR in mononuclear cells despite negativity of plasmatic PCR was noted 18 times in 8 patients. Persistently positive PCR in mononuclear cells in absence of detectable viraemia was followed by a virological relapse in 5 of these patients. This study confirms that hepatitis C virus RNA can be detected in mononuclear cells despite negative plasmatic PCR in patients treated by interferon. Moreover, the persistence of viral RNA in peripheral mononuclear cells could be a predictive factor of treatment failure. Our data also suggest that detection of viral RNA in mononuclear cells is probably not only due to passive virus adsorption from plasma.

关键词：抗病毒药/治疗应用随访研究基因型肝炎病毒属/药物作用肝炎病毒属/遗传学肝炎丙型/血液肝炎丙型/治疗干扰素类/治疗应用白细胞单核/药物作用白细胞单核/病毒学 RNA 病毒/血液 RNA 病毒/药物作用逆转录聚合酶链反应时间因素病毒血症/遗传学成年人女(雌)性人类男(雄)性中年人

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Treatment of chronic viral hepatitis

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ACTA CLINICA BELGICA 1999年第5期54卷 281-288页

作者： Henrion, J Hop Jolimont Serv Gastroenterol Dept Med Interne B-7100 Haine St Paul Belgium

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关键词：抗病毒药/治疗应用糖皮质激素类/治疗应用肝炎病毒属/药物作用肝炎病毒属/生理学乙型肝炎病毒/药物作用乙型肝炎病毒/生理学乙型肝炎慢性/药物疗法乙型肝炎慢性/病理生理学丙型肝炎慢性/药物疗法丙型肝炎慢性/病理生理学干扰素α/治疗应用干扰素类/治疗应用病人遴选泼尼松/治疗应用逆转录酶抑制剂/治疗应用利巴韦林/治疗应用人类

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