Management of hepatitis C virus infection in HIV/HCV co-infected patients: Clinical review

Nearly one fourth of individuals with human immunodeficiency virus (HIV) infection have hepatitis C virus (HCV) infection in the US and Western Europe. With the availability of highly active antiretroviral therapy and the consequent reduction in opportunistic infections, resulting in the prolongation of the life span of HIV-infected patients, HCV co-infection has emerged as a signif icant factor influencing the survival of HIV patients. Patients with HIV/HCV co-infection have a faster rate of fibrosis progression resulting in more frequent occurrences of cirrhosis, end-stage liver disease, and hepatocellular carcinoma. However, the mechanism of interaction between the two viruses is not completely understood. The treatment for HCV in co-infected patients is similar to that of HCV monoinfection; i.e., a combination of pegylated interferon and ribavirin. The presence of any barriers to antiHCV therapy should be identified and eliminated in order to recruit all eligible patients. The response to treatment in co-infected patients is inferior compared to the response in patients with HCV mono-infection. The sustained virologic response rate is only 38% for genotype-1 and 75% for genotype-2 and -3 infections. Liver transplantation is no longer considered a contraindication for end-stage liver disease in coinfected patients. However, the 5 year survival rate is lower in co-infected patients compared to patients with HCV mono-infection (33% vs 72%, P = 0.07). A better understanding of liver disease in co-infected patients is needed to derive new strategies for improving outcome and survival.

Shear-wave elastography to predict hepatocellular carcinoma after hepatitis C virus eradication:A systematic review and meta-analysis

BACKGROUND Direct-acting antiviral agents(DAAs)are highly effective treatment for chronic hepatitis C(CHC)with a significant rate of sustained virologic response(SVR).The achievement of SVR is crucial to prevent additional liver damage and slow down fibrosis progression.The assessment of fibrosis degree can be performed with transient elastography,magnetic resonance elastography or shear-wave elastography(SWE).Liver elastography could function as a predictor for hepato-cellular carcinoma(HCC)in CHC patients treated with DAAs.AIM To explore the predictive value of SWE for HCC development after complete clearance of hepatitis C virus(HCV).METHODS A comprehensive literature search of clinical studies was performed to identify the ability of SWE to predict HCC occurrence after HCV clearance.In accordance with the study protocol,a qualitative and quantitative analysis of the evidence was planned.RESULTS At baseline and after 12 wk of follow-up,a trend was shown towards greater liver stiffness(LS)in those who go on to develop HCC compared to those who do not[baseline LS standardized mean difference(SMD):1.15,95%confidence interval(95%CI):020-2.50;LS SMD after 12 wk:0.83,95%CI:0.33-1.98].The absence of a statistically significant difference between the mean LS in those who developed HCC or not may be related to the inability to correct for confounding factors and the absence of raw source data.There was a statist-ically significant LS SMD at 24 wk of follow-up between patients who developed HCC vs not(0.64;95%CI:0.04-1.24).CONCLUSION SWE could be a promising tool for prediction of HCC occurrence in patients treated with DAAs.Further studies with larger cohorts and standardized timing of elastographic evaluation are needed to confirm these data.

Liver biopsy in the post-hepatitis C virus era in Japan

In Japan,liver biopsies were previously crucial in evaluating the severity of hepatitis caused by the hepatitis C virus(HCV)and diagnosing HCV-related hepatocellular carcinoma(HCC).However,due to the development of effective antiviral treatments and advanced imaging,the necessity for biopsies has significantly decreased.This change has resulted in fewer chances for diagnosing liver disease,causing many general pathologists to feel less confident in making liver biopsy diagnoses.This article provides a comprehensive overview of the challenges and potential solutions related to liver biopsies in Japan.First,it highlights the importance of considering steatotic liver diseases as independent conditions that can coexist with other liver diseases due to their increasing prevalence.Second,it emphasizes the need to avoid hasty assumptions of HCC in nodular lesions,because clinically diagnosable HCCs are not targets for biopsy.Third,the importance of diagnosing hepatic immune-related adverse events caused by immune checkpoint inhibitors is increasing due to the anticipated widespread use of these drugs.In conclusion,pathologists should be attuned to the changing landscape of liver diseases and approach liver biopsies with care and attention to detail.

Autoimmune hepatitis and primary sclerosing cholangitis after direct-acting antiviral treatment for hepatitis C virus:A case report

BACKGROUND Chronic hepatitis C virus(HCV)infection is a major global health concern that leads to liver fibrosis,cirrhosis,and cancer.Regimens containing direct-acting antivirals(DAAs)have become the mainstay of HCV treatment,achieving a high sustained virological response(SVR)with minimal adverse events.CASE SUMMARY A 74-year-old woman with chronic HCV infection was treated with the DAAs ledipasvir,and sofosbuvir for 12 wk and achieved SVR.Twenty-four weeks after treatment completion,the liver enzyme and serum IgG levels increased,and antinuclear antibody became positive without HCV viremia,suggesting the development of autoimmune hepatitis(AIH).After liver biopsy indicated AIH,a definite AIH diagnosis was made and prednisolone was initiated.The treatment was effective,and the liver enzyme and serum IgG levels normalized.However,multiple strictures of the intrahepatic and extrahepatic bile ducts with dilatation of the peripheral bile ducts appeared on magnetic resonance cholangiopancreatography after 3 years of achieving SVR,which were consistent with primary sclerosing cholangitis.CONCLUSION The potential risk of developing autoimmune liver diseases after DAA treatment should be considered.

Ensemble for evaluating diagnostic efficacy of non-invasive indices in predicting liver fibrosis in untreated hepatitis C virus population

BACKGROUND Hepatitis C virus(HCV)infection progresses through various phases,starting with inflammation and ending with hepatocellular carcinoma.There are several invasive and non-invasive methods to diagnose chronic HCV infection.The invasive methods have their benefits but are linked to morbidity and complications.Thus,it is important to analyze the potential of non-invasive methods as an alternative.Shear wave elastography(SWE)is a non-invasive imaging tool widely validated in clinical and research studies as a surrogate marker of liver fibrosis.Liver fibrosis determination by invasive liver biopsy and non-invasive SWE agree closely in clinical studies and therefore both are gold standards.AIM To analyzed the diagnostic efficacy of non-invasive indices[serum fibronectin,aspartate aminotransferase to platelet ratio index(APRI),alanine aminotransferase ratio(AAR),and fibrosis-4(FIB-4)]in relation to SWE.We have used an Artificial Intelligence method to predict the severity of liver fibrosis and uncover the complex relationship between non-invasive indices and fibrosis severity.METHODS We have conducted a hospital-based study considering 100 untreated patients detected as HCV positive using a quantitative Real-Time Polymerase Chain Reaction assay.We performed statistical and probabilistic analyses to determine the relationship between non-invasive indices and the severity of fibrosis.We also used standard diagnostic methods to measure the diagnostic accuracy for all the subjects.RESULTS The results of our study showed that fibronectin is a highly accurate diagnostic tool for predicting fibrosis stages(mild,moderate,and severe).This was based on its sensitivity(100%,92.2%,96.2%),specificity(96%,100%,98.6%),Youden’s index(0.960,0.922,0.948),area under receiver operating characteristic curve(0.999,0.993,0.922),and Likelihood test(LR+>10 and LR-<0.1).Additionally,our Bayesian Network analysis revealed that fibronectin(>200),AAR(>1),APRI(>3),and FIB-4(>4)were all strongly associated with patients who had severe fibrosis,with a 100% probability.CONCLUSION We have found a strong correlation between fibronectin and liver fibrosis progression in HCV patients.Additionally,we observed that the severity of liver fibrosis increases with an increase in the non-invasive indices that we investigated.

Relationship of hepatitis C virus infection with primary hepatic carcinoma: an investigation of serum HCV RNA in different population groups

A preliminary study on the serum hepatitis C virus(HCV)RNA in 182 pa-tients with different chronic liver diseases and 35 blood donors was carried out with“nested”polymerase chain reaction.It was found that serum HCV RNA was detected in36.6%(34/93)cases of primary hepatic carcinoma(PHC),31.7%(20/63)liver cirrhosis,15.4%(4/26)chronic hepatitis and 2.9%(1/35)blood donors;most of the HCV RNA pos-itive cases(41/58)were complicated with HBV replicating markers;the rate of single posi-tive for HCV RNA was 15.1%(14/93)in PHC,3.2%(2/63)in liver cirrhosis,and 3.8%(1/26)in chronic hepatitis.These findings imply that about 20% of PHC cases appear tobe related to the coinfection of HCV and HBV and 15% of PHC cases are related to sin-gle HCV infection.

STUDY ON THE MOTHER TO INFANT TRANSMISSIONOF HEPATITIS C VIRUS BY COMBINED ASSAYOF ANTI-HCV AND HCV-RNA

objective The mother to infant transmission of Hey was prospectively investigated.Methods Hepatitis C virus(HCV) infection was tested by the combination assay of anti-HCV andHCV- RNA. Six hundred and ten pregnant women were investigated for HCV infection, and infants from HCVinfected mothers were followed up at birth, 3, 6, 9, 12 months after birth to investigate HCV infection. HCVgenotypes were detected in all persons with HCV- RNA positive. HCV was quantified by branch DNA signalamplification assay (bDNA) in pregnant women. Results Among 610 pregnant women, 18 infected HCV, theinfection rate of HCV in pregnant women was 2.95% (18/610). Five of 18 infants from 18 HCV infected mothersinfected HCV, they had no history of operation, or blood transfusion and other risk exposure to HCV, so the HCVinfection was irc m their mothers, the rate of HCV transmission from mother to infant was 27.8%.HCVgenotype fo was found in 16 pregnant women with HCV- RNA positive, 5 infants and their mothers had the sameHCV genotype(1b) infection. All pregnant women infected HCV have low HCV titer in serum. Conclusion lnthis research, it was noted that HCV could be vertically transmitted from mother to infant even if mother had lowserum HCV titer(≤14.11 ×105/ml). The combination assay of anti - HCV and HCV- RNA was valuable ininvestigating the HCV infection in pregnant women and the transmission of HCV from mother to infant. It haspotential value in diagnosing HCV infection in other population.

Lichen planus and hepatitis c virus (HCV)-there is no association:A serological case control study

Fifty patients(28 males and 22 females)with lichen planus were tested for anti HCV antibodies.None of them gave positive result in our case control study.There were various studies conducted in different parts of the world that proved or disproved a causative role for HCV in LP.

Hepatitis C virus micro-elimination:Where do we stand?

Hepatitis C virus (HCV) elimination by 2030, using direct-acting antiviraltreatments, has been promoted by the World Health Organization. Thisachievement is not attainable, however, particularly after the 2020 pandemic ofthe coronavirus disease 2019. Consequently, the more realistic objective ofeliminating HCV from population segments for which targeted strategies ofprevention and treatment are easily attained has been promoted in Europe, as avalid alternative. The underlying idea is that micro-elimination will ultimatelylead to macro-elimination. The micro-elimination strategy may target differentspecific populations and at-risk groups. Different settings, including prisons andhospitals, have also been identified as micro-elimination scenarios. In addition,dedicated micro-elimination strategies have been designed that are tailored at thegeographical level according to HCV epidemiology and individual country’sincome. The main elements of a valid and successful micro-elimination project arereliable epidemiological data and active involvement of all the stakeholders.Community involvement represents another essential component for a successfulprogram.

慢性丙型肝炎合并非酒精性脂肪肝患者血清25(OH)D水平变化及其与HCV-RNA载量、糖脂代谢的关系

目的 探讨慢性丙型肝炎(CHC)合并非酒精性脂肪性肝病(NAFLD)患者血清25-羟维生素D[25(OH)D]水平及其与HCV-RNA载量、糖脂代谢的关系。方法 选取2018年9月至2021年9月医院收治CHC患者174例,根据是否合并NAFLD分为单纯CHC组(110例)和CHC-NAFLD组(64例),选取同期体检健康成人87例(健康组),使用酶联免疫吸附法检测纳入对象血清25(OH)D水平,生化分析检测糖脂代谢指标,荧光定量PCR法检测CHC患者HCV-RNA载量,spearman相关性分析血清25(OH)D水平与HCV-RNA载量、糖脂代谢指标水平的相关性,logistics回归分析CHC合并NAFLD的危险因素。结果 CHC-NAFLD组、CHC组患者中BMI≥28kg/m^(2)比率、HbA1c、FBG、INS、IR、TG和LDL-C水平显著高于健康组(P<0.05),血清25(OH)D水平显著低于健康组(P<0.05)。CHC-NAFLD组BMI≥28kg/m^(2)比率、HbA1c、FBG、INS、IR、TG、LDL-C水平和HCV-RNA载量显著高于CHC组(P<0.05),血清25(OH)D水平显著低于CHC组(P<0.05)。25(OH)D水平与HbA1c、FBG、IR、TG水平呈负相关(r=-0.426、-0.387、-0.406、-0.514,P<0.001)。25(OH)D<42 nmol/L、BMI≥28 kg/m^(2)、TG≥1.72 mmol/L、HCV-RNA载量≥3.5×10^(6) IU/mL是CHC患者合并NAFLD的危险因素(P<0.05)。结论 CHC合并NAFLD患者血清25(OH)D水平降低,25(OH)D水平与HbA1c、FBG、IR、TG呈负相关且其水平升高会增加CHC患者发NAFLD发生风险。

Glecaprevir/pibrentasvir+sofosbuvir for post-liver transplant recurrent hepatitis C virus treatment

Glecaprevir/pibrentasvir in combination with sofosbuvir may serve as a safe and effective option for treatment of recurrent hepatitis C virus post-liver transplant in patients who previously failed direct-acting antivirals.

Hepatitis C virus treatment with glecaprevir and pibrentasvir in patients co-prescribed carbamazepine:Three case reports

BACKGROUND Highly effective and well-tolerated direct-acting antiviral(DAA)therapies have revolutionised the management of hepatitis C virus(HCV);however,niche populations face treatment barriers.DAAs co-prescribed with several firstgeneration anti-epileptic drugs(AEDs)are contraindicated due to drug-drug interactions.A common example is carbamazepine whereby steady-state carbamazepine reduces the maximum concentration and area under the curve of velpatasvir,glecaprevir and pibrentasvir due to potent cytochrome P450(CYP)3A4 induction.Carbamazepine also induces P-glycoprotein which reduces glecaprevir and pibrentasvir’s area under curve to infinite time.Sofosbuvirvelpatasvir and glecaprevir-pibrentasvir are contraindicated in patients who are co-prescribed carbamazepine due to the risk of reduced DAA therapeutic effect and consequently,virological treatment failure.This presents a challenge for patients in whom carbamazepine substitution is medically unfeasible,impractical or unacceptable.However,the properties of current generation DAA therapies,including high-potency non-structural protein 5A inhibitory effect,may be sufficient to overcome reduced bioavailability arising from carbamazepine related CYP 3A4 and P-glycoprotein induction.CASE SUMMARY We present a case series of three patients with non-cirrhotic,treatment-naïve,genotype 1a,1b,and 3a HCV who were treated with a 12 wk course of glecaprevir-pibrentasvir,while co-prescribed carbamazepine for seizure disorders.Glecaprevir-pibrentasvir combination therapy was chosen due to its potent in vitro activity and low barrier to pan-genotypic resistance associated variants.DAA therapy was dose-separated from carbamazepine to maximise time to peak concentration,and taken with meals to improve absorption.Sustained virological response at 12 wk was achieved in each patient with no adverse outcomes.CONCLUSION DAA therapies,including glecaprevir-pibrentasvir,warrant consideration as a therapeutic agent in people with HCV who are co-prescribed carbamazepine,particularly if AED substitution is not feasible.

Prevalence of HCV Antibody and its Associated Factors: A Study from Sentinel Hospitals in China

Objective The prevalence and related factors of serum anti-HCV in different regions and hospitals have not been studied extensively in China. We used routine screening data to determine the prevalence of HCV antibody in hospital patients, evaluate the epidemic trend of hepatitis C and formulate screening strategies.Methods Patient information and HCV antibody testing results were collected from January 2017 to December 2019 in 77 HCV sentinel hospitals in China. Univariate and multivariate logistic regression was used to determine the characteristics and associations.Results HCV antibody prevalence rates were distinct among patients in different departments, with a range of 0.33%–6.93%. Patients who were admitted to the liver disease-related departments(aOR =10.76;95% CI, 10.27–11.28), Internal Medicine(aOR = 2.87;95% CI, 2.75–3.00), and Department of Surgery(aOR = 1.95;95% CI, 1.87–2.04), were more likely to be tested for HCV antibody positive. HCV antibody prevalence was associated with patients aged 45 years and older(aOR = 2.74;95% CI,2.69–2.80), testing in infetious disease hospitals(aOR = 2.33;95% CI, 2.26–2.40) and secondary hospitals(aOR = 1.72;95% CI, 1.69–1.75). Patients in sentinel hospitals of the Northeast(aOR = 12.75;95% CI,12.40–13.11), the Central(aOR = 1.65;95% CI, 1.61–1.70), and the West(aOR = 1.78;95% CI, 1.73–1.83)China had higher HCV prevalence than those who were in the Eastern coastal area. Conclusion Those who were over 45 years old and saw doctors for liver diseases, and invasive diagnosis and treatment should be referred to HCV antibody testing.

丙型病毒性肝炎抗体检测与HCV高敏核酸检测对病毒性丙型肝炎的筛查价值研究

目的通过对本院2019年1月至2022年9月需进行手术、输血或其他侵入性医疗服务的住院患者的HCV血清学抗体及高敏HCV RNA筛查结果进行回顾性分析,探讨丙型病毒性肝炎抗体检测与HCV高敏核酸检测对病毒性丙型肝炎的筛查检测价值。方法对本院相关住院患者进行Elecsys Anti-HCV II或(和)高敏HCV RNA检测,统计分析其检测结果。结果HCV血清学抗体检测22443人次,阳性率为0.68%,阳性COI中位数为38.4(1.0~165)。高敏HCV RNA检测34628人次,阳性率0.30%,阳性病毒载量中位数为1.00×10^(6)IU/mL(3.50×10^(1)~4.00×10^(7)IU/mL)。两种方法学均显示45~59岁人群阳性率显著高于其他人群(P<0.001)。两种检测有重合的样本共17785人次,HCV抗体血清学阳性率为0.70%。高敏HCV RNA阳性率0.22%,如以高敏HCV RNA为丙型肝炎现症感染的标准,HCV抗体血清学检测HCV现症感染的灵敏度为100.00%(95%CI 89.09%~100.00%),特异性为99.52%(95%CI 99.41%~99.61%),现症感染阳性预测期PPV为32.00%(95%CI 23.82%~40.18%),阴性预期值为100.00%。HCV抗体血清学检测COI值与高敏HCV RNA检测的病毒载量无线性相关性,COI<10和COI>100的HCV RNA阳性率低。结论HCV抗体检测和高敏HCV RNA均为有效的丙型肝炎筛查手段,需进一步加强对重点人群的丙型肝炎感染管理。

格卡瑞韦/哌仑他韦治疗HCV/HIV合并感染患者的临床研究

目的分析格卡瑞韦/哌仑他韦治疗丙型肝炎病毒(HCV)/人类免疫缺陷病毒(HIV)合并感染患者的临床效果和安全性,以期为临床治疗提供科学依据。方法选择2021年1月—2022年1月凉山州布拖县某医院收治的89例HCV/HIV合并感染无肝硬化的初治患者,均予以8周的格卡瑞韦/哌仑他韦治疗,随访12周。观察并记录治疗结束时的病毒学应答率、治疗结束12周后的持续病毒性应答率(SVR12)及不良反应发生情况。结果89例HCV/HIV合并感染无肝硬化的初治患者多为中青年已婚男性(79例,88.8%),HIV感染主要经性接触传播(62例,69.7%)和静脉注射毒品传播(27例,30.3%)。HCV基因型最常见的是基因1b型(33例,37.1%)和基因3b型(25例,28.1%)。全部患者均顺利完成8周治疗,且治疗结束时HCV RNA载量均低于检测值下限(<25 IU/mL)。其中,8例患者未能完成随访,余81例(100%)患者均获得持续病毒学应答。患者观察期间均未出现严重不良反应,但有11例患者发生轻度不良反应。结论格卡瑞韦/哌仑他韦8周方案治疗基因1、3、6型HCV/HIV合并感染无肝硬化的初治患者SVR12达100%,且安全性和耐受性均较好,可以作为此类患者临床治疗的优先选择。

HCV不同标志物检测结果比较分析

目的分析丙型肝炎病毒抗原、抗体及核酸标志物实验室检测结果之间的关联性。方法用丙型肝炎病毒抗体(HCV-Ab)、丙型肝炎病毒核酸(HCV-RNA)扩增(PCR)荧光定量、丙型肝炎病毒核心抗原(HCV-CAg)及丙型肝炎病毒抗原(HCV-Ag)检测试剂盒,分别检测血清或血浆样品中的HCV-Ab、HCV-RNA、HCV-CAg和HCV-Ag丙型肝炎病毒4种标志物,对检测结果之间的关联性进行分析。结果在1551份血清或血浆样品中,共检出HCV-Ab阳性样品565份(36.43%)、HCV-Ag(游离抗原)阳性样品48份(3.09%)、HCV-RNA阳性样品317份(20.44%)、HCV-CAg阳性样品25份(1.61%),HCV-Ab及HCV-RNA检出率明显高于抗原检出率(P<0.01),其阳性样品检出率按高低顺序依次为HCV-Ab、HCV-RNA、HCV-Ag和HCV-CAg。结论 4种标志物的检测可以联合运用,能有效降低HCV-Ab检测带来的漏检风险。

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.

Innovations in education:A prospective study of storytelling narratives to enhance hepatitis C virus knowledge among substance users

BACKGROUND Even though substance users have the highest hepatitis C virus(HCV)burden,many lack knowledge about the infection.Lack of knowledge is an important obstacle to pursuing HCV care.Although printed materials are conventionally utilized to disseminate HCV-related knowledge,narrative story-telling videos may be an alternative.Data are extremely limited,however,in the ability of storytelling videos to increase HCV knowledge among substance users.In this study,we hypothesized that a story-telling narrative video would increase substance user’s immediate and 1-month HCV-related knowledge compared to a printed format.AIM To assess immediate and 1-month HCV-related knowledge retention among substance users comparing education delivered via a storytelling narrative video compared to a printed format.METHODS We conducted a prospective matched,case-control study among substance users actively prescribed buprenorphine enrolled from two sites.The intervention site received the video and the control site,the brochure.Participants(n=176)were matched on age,gender,and race.We obtained extensive patient and stakeholder input on the video’s design,validated the video’s content,and developed a recruitment plan to guide participant enrollment.Knowledge was assessed by administration of a 25-item instrument immediately before,immediately after,or one month after the intervention.Data were analyzed using nonparametric and generalized linear mixed-effects models.RESULTS We recruited a total of 176 substance users,90 and 86 individuals,from each site,respectively.One-month follow up occurred in 92%and 94%of enrollees in the control and intervention groups,respectively.In comparison with the pre-intervention scores,immediate knowledge recall increased significantly for both the intervention(P<0.0001)and control(P<0.0001)groups.Multivariate modeling revealed a significant improvement in HCV-related knowledge and retention(P=0.033)among participants who viewed the storytelling video.CONCLUSION Storytelling narratives emphasizing HCV education appear to be an effective method to increase HCV-related knowledge among substance users.They should become an educational cornerstone to promote HCV management among this population.

S-adenosyl-L-methionine modifies antioxidant-enzymes,glutathione-biosynthesis and methionine adenosyltransferases-1/2 in hepatitis C virus-expressing cells

AIM: To elucidate the mechanism(s) by which S-adenosyl-L-methionine(SAM) decreases hepatitis C virus(HCV) expression.METHODS: We examined the effects of SAM on viral expression using an HCV subgenomic replicon cell culture system. Huh7 HCV-replicon cells were treated with 1 mmol/L SAM for different times(24-72 h), then total RNA and proteins were isolated. c DNA was synthesized and real time-PCR was achieved to quantify HCV-RNA, superoxide dismutase 1 and 2(SOD-1, SOD-2) catalase, thioredoxin 1, methionine adenosyltransferase 1A and 2A(MAT1A, MAT2A) expression, and GAPDH and RPS18 as endogenous genes. Expression of cellular and viral protein was evaluated by western-blot analysis using antibodies vs HCV-NS5 A, SOD-1, SOD-2, catalase, thioredoxin-1, MAT1 A, MAT2 A, GAPDH and actin. Total glutathione levels were measured at different times by Ellman's recycling method(0-24 h). Reactive oxidative species(ROS) levels were quantified by the dichlorofluorescein assay(0-48 h); Pyrrolidin dithiocarbamate(PDTC) was tested as an antioxidant control and H2O2 as a positive oxidant agent.RESULTS: SAM exposition decreased HCV-RNA levels 50%-70% compared to non-treated controls(24-72 h). SAM induced a synergic antiviral effect with standard IFN treatment but it was independent of IFN signaling. In addition, 1 mmol/L SAM exposition did not modify viral RNA stability, but it needs cellular translation machinery in order to decrease HCV expression. Total glutathione levels increased upon SAM treatment in HCV-replicon cells. Transcriptional antioxidant enzyme expression(SOD-1, SOD-2 and thioredoxin-1) was increased at different times but interestingly, there was no significant change in ROS levels upon SAM treatment, contrary to what was detected with PDTC treatment, where an average 40% reduction was observed in exposed cells. There was a turnover from MAT1A/MAT2 A, since MAT1 A expression was increased(2.5 fold-times at 48 h) and MAT2 A was diminished(from 24 h) upon SAM treatment at both the transcriptional and translational level. CONCLUSION: A likely mechanism(s) by which SAM diminish HCV expression could involve modulating antioxidant enzymes, restoring biosynthesis of glutathione and switching MAT1/MAT2 turnover in HCV expressing cells.

Treatment of hepatitis C virus infection

Acute and chronic hepatitis C virus (HCV) infection remains a serious health problem worldwide, however, there has been advancement in the treatment of HCV infection due to standard treatment using pegylated interferon and ribavirin. The literature indicates that therapy for HCV is becoming more individualized. In addition to considering genotype and viral RNA levels before treatment, achievement of an early virologic response (EVR) and a rapid virologic response (RVR) is now possible during therapy. Moreover, problem patients, such as non-responders, relapsers, HIV or HBV co- infected patients, patients with liver cirrhosis, and pre- or post-liver transplantation patients are an increasing fraction of the patients requiring treatment. This article reviews the literature regarding standard treatments and problem patients with acute and chronic HCV infection. It also includes discussion on contraindications and side effects of treatment with interferon and ribavirin, as well as new drug development.