广西肝病患者及献血员的HCV Serotype和Genotype的分子生物学研究

选择广西不同地市的肝病患者１０３例（ＡＨ８、ＣＡＨ３６、ＣＰＨ３６、ＬＣ７、ＨＣＣ１６）和献血员３５５例进行Ａｎｔｉ－ＨＣＶ阳性率、ＨＣＶＳｅｒｏｔｙｐｅ和Ｇｅｎｏｔｙｐｅ的分子生物学研究。结果表明：肝病患者Ａｎｔｉ－ＨＣＶ阳性１２例（ＡＨ１／８。ＣＡＨ２／３６。ＣＰＨ９／３６。ＬＣＯ／７、ＨＣＣ０／１６），阳性率为１１．６５％，明显高于同室检测兰州和南京肝病患者的Ａｎｔｉ－ＨＣＶ阳性率４．３％和４．２％（Ｐ＜０．０１）；２４０例献血员中检出Ａｎｔｉ－ＨＣＶ阳性１１例（４．５８％），明显低于同室检测兰州献血员的Ａｎｔｉ－ＨＣＶ阳性率３５．００％（Ｐ＜０．０１）；本文两组的Ａｎｔｉ－ＨＣＶ阳性率也存在显著性差异（Ｐ＜０．０１）。肝病患者组的ＨＣＶＳｅｒｏｔｐｙｅ１型、２型、１＋２型和无法定为１和／或２型者分别为３３．３３％、０、０和６６．６７％；而献血员组的ＨＣＶＳｅｒｏｔｙｐｅ分别为６５．９５％、１．１０％、２．２０％和３０．７７％。此外，肝病患者组的ＨＣＶＧｅｎｏｔｙｐｅ均属ｌｂ型（５０．００％）和ｌｂ混合型（５０．００％）；献血员组的ＨＣＶＧｅｎｏｔｙｐｅ中ｌａ型、ｌｂ型、ｌｂ混合型和无法定为１和／或２?

HCV患者年龄、抗体和HCV RNA与血清分型的关系

目的采用建立的丙型肝炎病毒（HCV）血清分型技术,探讨HCV患者年龄、抗体（S/CO）和HCV RNA与血清分型的关系。方法收集HCV慢性感染患者血清200例,定量检测HCV RNA、抗-HCV,同时进行血清分型,并进行统计分析。结果 200例慢性丙肝患者的血清抗-HCV抗体均为阳性,S/CO〉3.5,HCV RNA滴度在1.12×103~9.29×107 copies/ml。其中血清1型102例,2型58例,混合型2例,分型率为81.00%。采用Student t检验分析年龄、抗体S/CO、HCV RNA与分型之间的关系,t值分别为1.842 9,0.446 7,1.778 2,P均〉0.05,无统计学意义。结论 HCV患者年龄、抗体（S/CO）和HCV RNA滴度对血清分型结果没有影响。

Phytochemical analysis of Berberis lyceum methanolic extract and its antiviral activity through the restoration of MAPK signaling pathway modulated by HCV NS5A

Objective:To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase(MAPK)signaling pathway modulated by hepatitis C virus(HCV)nonstructural protein 5 A(NS5A).Methods:A total of ten plant extracts were initially screened for their toxicities against Hep G2 cells.The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both m RNA and protein levels using real-time PCR and Western blotting assays,respectively.The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR.Subsequently,the identification of secondary metabolites was carried out by phytochemical and HPLC analysis.Results:The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids,phenols,saponins,tannins,flavonoids,carbohydrates,terpenoids,steroids,and glycosides.Similarly,quercetin,myricetin,gallic acid,caffeic acid,and ferulic acid were identified through HPLC analysis.The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44μg/m L.RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dosedependent manner.Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway.Conclusions:Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.

CHANGES OF ANTIBODIES AGAINST ANTIGENS ENCODED BY DIFFERENT REGIONS OF HCV GENOME IN CHRONIC HEPATITIS C PATIENTS TREATED WITH INTERFERON

Twenty patients with chronic hepatitis c were investigated during the treatment with interferon to explore the changes of antibodies to HCV (anti-RCV). Anti-HCV was tested with recombinant immunoblot assay (RIBA) by using three antigens (C22, C33c, and C100-3) encoded by different regions of HCV genome. The changes of individual anti-HCV and ALT were compared with the change of HCV RNA. The results showed that persistent disappearance of serum HCV RNA was closely related to the changes of anti-C33c (P<0. 01) and anti-C100-3 (P<0. 005), but there was no relation between persistent ALT normality and HCV viremia clearance (P<0. 05). In conclusion, monitoring anti-C33c and anti-C100-3 could indicate the changes or HCV viremia. The normalization of ALT after interferon treatment did not indicate disappearance of HCV viremia.

山东省部分地区献血员丙型肝炎病毒感染状况及分型研究

选择 42 4例有偿献血员用ELISA检测抗 -HCV ,对抗 -HCV阳性血清用ELISA进行血清学分型 ,对己确定血清型者采用RT—PCR作基因分型。结果 42 4例献血员中 ,HCV感染率为 2 1 70 % ( 92 / 4 2 4) ,献全血并血浆者的HCV感染率( 45 14% )显著高于只献全血者 ( 6 94% ) ;92例抗—HCV阳性者中 ,血清学分型率为 34 78% ( 32份 ) ,其中血清学Ⅰ型 30例( 93 75 % )、Ⅱ型 2例 ( 6 2 5 % ) :该 32例血清中有 30例可用基因分型 ,其中基因Ⅱ型 2 6例 ( 86 6 7% )、Ⅲ型 3例 ( 10 % )、Ⅱ /Ⅲ型混合感染 1例 ( 3 33% ) ,两种分型方法的一致率为93 33 % ( 2 8/ 30 )。表明山东省部分地区献血人群的HCV感染率较高 ;献血浆是该献血人群HCV感染的危险因素 ;

广西吸毒成瘾者丙型肝炎病毒的感染及其分子生物学研究

选择２８３名静脉吸毒者（ＩＶＤＡｓ）和１２１名献血员（ＢＤｓ）进行Ａｎｔｉ－ＨＣＶ、ＨＣＶ血清基因型、ＨＣＶ基因型和ＨＣＶｃＤＮＡ序列的检测。结果表明，ＩＶＤＡｓ和ＢＤｓ的Ａｎｔｉ－ＨＣＶ检出率分别为９１．１７％和０．８３％；ＩＶＤＡｓ的ＨＣＶ血清基因型为１型８１．８５％（２２１／２７０），２型１．４８％（４／２７０），ｌ＋２型０．３７％（１／２７０），不能定为１和／或２型１６．３０％（４４／２７０）；ＨＣＶ基因型为１ａ型：２８．６％（３４／１１９）；ｌｂ型：３８．７％（４６／１１９）；２ａ型１０．９％（１３／ｌ１９）；２ｂ型１４．３％（１７／ｌ１９）；３ａ型２６．９％（３２／１１９）；３ｂ型４０．３％（４８／１１９）；６ａ型８．４％（１０／１１９）；６ｂ型２６．７％（３１／１１９）；其中１４．３％的病例有４～５种不同基因亚型的混合感染现象。

罗氏诊断新一代Elecsys anti-HCV Ⅱ检测试剂上市

近日，罗氏诊断新一代Elecsys anti-HcV Ⅱ检测试剂上市会在上海召开，来自临床肝病领域、传染病实验室检测领域的专家以及罗氏诊断总部的专家等做了大会演讲。与会专家表示，如能早筛查、早诊断，丙肝治疗效果很好，约70％～80％的患者可通过规范治疗实现治愈。但其高隐匿、高漏诊、高慢性化的特点，使许多患者错过了最佳治疗时机。

The Diagnostic Value of Anti-HCV Test in HCV Infection

Two hundred and ninety-six samples of patient serum and 28 samples of donor serum were tested for anti-HCV with second generation domestic made testing kit,and for HCV RNA with PCR assay.The purpose of the study was

Hepatitis C Status in Karachi, Pakistan, a Five-Year Survey at Civil Hospital, Karachi, Pakistan

Hepatitis C virus is associated with Hepatitis C which can be acute or chronic. Acute hepatitis C is usually short term illness that can lead to chronic infection, while chronic hepatitis C is a chronic liver disease leading to cirrhosis, hepatocellular carcinoma and even death. The mode of transmission is mainly parenteral and vertical. Hepatitis C is the leading cause of morbidity and mortality and a serious public health problem, worldwide as well as in Pakistan. There is no vaccine available for hepatitis C so the only way to prevent hepatitis C is by changing behaviors which can spread the disease. Pakistan is also facing a huge burden of this disease. In Pakistan, the frequency of hepatitis C virus infection ranges from 8% - 15% in the general population with variations in different parts of the country. The general method of detecting infection with HCV is to observe the presence of antibodies to the virus. The aim of this study was to determine frequency of hepatitis C among general population and to see the increasing frequency in our part. Selection of Patients: All patients visiting outpatient clinics, Hepatitis clinic and indoor patients at Civil Hospital Karachi, Pakistan from January 1, 2010 to December 31, 2014 were screened for anti HCV. Methods: Blood samples were collected from the patients in Red Topped vaccutainers and allowed to clot, then centrifuged and serum was screened for anti HCV by Immunochromatography (ICT) method. Results: A total of 156,990 patients were screened for anti-HCV, 23,511 (14.98%) patients were reported positive. During the year 2010, total 11,058 patients were screened for anti HCV, 1791 (16%) were positive, while in the year 2011 total 26,921 were screened and 3908 (14.5%) were positive, during the year 2012, total 23,321 were screened, 3036 (14%) were positive, in the year 2013 we screened total 40,483 patients 6070 (15%) were positive, while in 2014, total 55,484 patients were screened and 8584 were positive (15.5%). The most affected age group is between 21 - 40 years and males are affected more than females. Conclusion: Hepatitis C in increasing in our population affecting the young generation between the age of 21 - 40 years and male population is affected more than females so a mass screening is needed to know the exact position of the disease as well as awareness programs should be started using media.

丙型肝炎病毒血清学分型的研究

本文用EIA法对辽宁省部分地区的105例抗-HCV阳性者(包括各种丙型肝炎患者和无症状丙型肝炎病毒感染者)进行抗-HCV型特异性抗体检测,结果显示血清学方法的分型率为72.9％(76/105),其中血清1型44人(57.9％),血清2型29人(38.2％),1/2混合型占3.9％(3/76)。提示在辽宁省部分地区的抗-HCV阳性人群中HCV血清1、2型均存在,但以血清1型为主。

丙型肝炎病毒血清抗体分型研究

为探讨丙型肝炎病毒(HCV)血清抗体分型的可能性及其意义,采用酶联免疫技术(EIA)对140例丙型肝炎患者血清抗-HCV 型特异性抗体进行检测,并比较其与基因分型的检测结果。结果显示63.6%(89)的患者检测出 HCV 血清型特异性抗体,其中血清1型占87.6%(78/89),血清2型占6.7%(6/89),血清1+2型占5.6%(5/89)。对其中71例血清 HCV RNA 阳性者的血清型结果与基因型结果相比较,二者符合率达97.2%。研究表明 HCV 血清抗体分型简便易行,尤其可用于对部分抗-HCV 阳性而 HCV RNA 阴性患者进行分型。但其敏感性尚有待于提高。

慢性丙型肝炎干扰素抗体及血清分型与派罗欣疗效关系探讨

目的探讨慢性丙型肝炎抗_IFN及血清分型与派罗欣(聚乙二醇IFNα_2a注射液)疗效的关系。方法接受派罗欣治疗的慢性丙型肝炎患者32例,用ELISA法进行血清分型及抗_IFN的测定。结果32例慢性丙型肝炎患者中HCV血清1型、血清2型及血清1+2混合型阳性率分别为50.00%(16/32)、31.25%(10/32)及18.75%(6/32)。32例慢性丙型肝炎患者,总体抗_IFN检出率为53.13%(17/32),其中HCV血清1型、2型及1+2混合型阳性率分别为:31.25%(10/32)、15.63%(5/32)、6.25%(2/32),血清1型与2型相比较差异显著(P<0.05)。派罗欣治疗后,血清2型患者HCVRNA含量<103拷贝/ml,血清1型患者HCVRNA仍>103拷贝/ml。结论32例慢性丙型肝炎患者HCV血清1型为主;派罗欣治疗后血清2型效果优于血清1型。

Emerging roles of interferon-stimulated genes in the innate mmune response to hepatitis C virus infection

Infection with hepatitis C virus （HCV）, a major viral cause of chronic liver disease, frequently progresses to steatosis and cirrhosis, which can lead to hepatocellular carcinoma. HCV infection strongly induces host responses, such as the activation of the unfolded protein response, autophagy and the innate immune response. Upon HCV infection, the host induces the interferon （IFN）-mediated frontline defense to limit virus replication. Conversely, HCV employs diverse strategies to escape host innate immune surveillance. Type I IFN elicits its antiviral actions by inducing a wide array of IFN-stimulated genes （ISGs）. Nevertheless, the mechanisms by which these ISGs participate in IFN-mediated anti-HCV actions remain largely unknown. In this review, we first outline the signaling pathways known to be involved in the production of type I IFN and ISGs and the tactics that HCV uses to subvert innate immunity. Then, we summarize the effector mechanisms of scaffold ISGs known to modulate IFN function in HCV replication. We also highlight the potential functions of emerging ISGs, which were identified from genome-wide siRNA screens, in HCV replication. Finally, we discuss the functions of several cellular determinants critical for regulating host immunity in HCV replication. This review will provide a basis for understanding the complexity and functionality of the pleiotropic IFN system in HCV infection. Elucidation of the specificity and the mode of action of these emerging ISGs will also help to identify novel cellular targets against which effective HCV therapeutics can be developed.

Role of Ras-related Nuclear Protein/Polypyrimidine Tract Binding Protein in Facilitating the Replication of Hepatitis C Virus

Background and Aims:Ras-related nuclear(RAN)protein is a small GTP-binding protein that is indispensable for the translocation of RNA and proteins through the nuclear pore complex.Recent studies have indicated that RAN plays an important role in virus infection.However,the role of RAN in hepatitis C virus(HCV)infection is unclear.The objective of this study was to investigate the role and underlying mechanisms of RAN in HCV infection.Methods:Huh7.5.1 cells were infected with the JC1-Luc virus for 24 h and then were incubated with complete medium for an additional 48 h.HCV infection and RAN expression were determined using luciferase assay,quantitative reverse transcription-PCR and western blotting.Small interfering RNA was used to silence RAN.Western blotting and immunofluorescence were used to evaluate the cytoplasmic translocation of polypyrimidine tract-binding(PTB),and coimmunoprecipitation was used to examine the interaction between RAN and PTB.Results:HCV infection significantly induced RAN expression and cytoplasmic redistribution of PTB.Knockdown of RAN dramatically inhibited HCV infection and the cytoplasmic accumulation of PTB.Colocalization of RAN and PTB was determined by immunofluorescence,and a direct interaction of RAN with PTB was demonstrated by coimmunoprecipitation.Conclusions:PTB in the host cytoplasm is directly associated with HCV replication.These findings demonstrate that the involvement of RAN in HCV infection is mediated by influencing the cytoplasmic translocation of PTB.