Blood micronutrient, oxidative stress, and viral load in patients with chronic hepatitis C

AIM: To assess the extent of micronutrient and oxidative stress in blood and to examine their linkages with viral loads in chronic hepatitis C patients. METHODS: Hepatitis C virus (HCV)-RNA levels were quantified in the serum from 37 previously untreated patients with chronic hepatitis C. The plasma and erythrocyte micronutrients (zinc, selenium, copper, and iron) were estimated, and malondialdehyde (MDA) contents were determined as a marker to detect oxidative stress. Antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione reductase (GR) activities in blood were also measured. The control group contained 31 healthy volunteers. RESULTS: The contents of zinc (Zn), and selenium (Se) in plasma and erythrocytes were significantly lower in hepatitis C patients than in the controls. On the contrary, copper (Cu) levels were significantly higher. Furthermore, plasma and erythrocyte MDA levels, and the SOD and GR activities in erythrocytes significantly increased in hepatitis C patients compared to the controls. However, the plasma GPX activity in patients was markedly lower. Plasma Se (R=-0.730, P<0.05), Cu (r=0.635), and GPX (r= -0.675) demonstrated correlations with HCV-RNA loads. Significant correlation coefficients were also observed between HCV-RNA levels and erythrocyte Zn (r=-0.403), Se (r = -0.544), Cu (r= 0.701) and MDA (P=0.629) and GR (r =0.441). CONCLUSION: The levels of Zn, Se, Cu, and oxidative stress (MDA), as well as related anti-oxidative enzymes (GR and GPX) in blood have important impact on the viral factors in chronic hepatitis C. The distribution of these parameters might be significant biomarkers for HCV.     

Cytogenetic methods for detection of oxidative stress and evaluation of antioxidant therapy in hepatitis C infection

The plasma of patients with hepatitis C contains chromosome-damaging substances, the so-called "clastogenic factors" (CFs), as this is the case for other chronic inflammatory diseases and after radiation exposure. These endogenous clastogens, formed as a consequence of increased superoxide production by inflammatory cells, can be detected with cytogenetic methods, as they are used for exogenous clastogens. The long-lived, autosustained DNA-damaging effects of CFs are risk factors for the development of cancer and leukemia. In hepatitis C, the highest clastogenic scores has been observed in patients with hepatocellular carcinoma. In agreement with the link to inflammation, clastogenic score are correlated with necro-inflammatory scores in liver biopsies. Antioxidant therapy with a powerful superoxide scavenger resulted in normalization of clastogenic scores and significant decreases in aminotransferase levels, but did not influence the virus load. Preliminary results of our study on a limited number of patients suggest that pre-treatment with antioxidants may improve the outcome of interferon/ribavirin treatment. A comparison of a three-month treatment with either interferon alone or the antioxidant alone, yielded similar results for reduction of ALT levels, but only complete normalization of clastogenic scores for the antioxidant. Further studies have to be conducted to see whether a combination of an antiviral agent with an appropriate antioxidant would allow to reduce interferon and its side effects.Combination of antioxidants with IFN/RIBA was also reported by other authors with discordant results. The CF-test can be useful in clinical trials for the choice of the appropriate antioxidant.     

Challenge of hepatitis C in Egypt and hepatitis B in Mauritania

Egypt has one of the highest prevalence rates of hepatitis C virus(HCV) in the world,mostly with genotype 4 that is highly associated with severe fibrosis. As a consequence,hepatocellular carcinoma has become the leading cause of cancer in this country. Mauritania is a highly endemic area for hepatitis B virus(HBV). HBV and HCV could both be iatrogenically transmitted through infected blood products,infected needles,and medical equipment improperly sterilized. Adequate and efficient healthcare and public health measures with good surveillance programs,access for screening,prevention strategies,and successful treatment are needed to halt the spread of these diseases. Herein,we have reviewed the epidemiology,modes of transmission,predisposing factors,and novel treatment modalities of these viruses. We have proposed practices and interventions to decrease the risk of transmission of HCV and HBV in the affected countries,including strict adherence to standard precautions in the healthcare setting,rigorous education and training of patients and healthcare providers,universal screening of blood donors,use of safetyengineered devices,proper sterilization of medical equipment,hepatitis B vaccination,as well as effective direct-acting antiviral agents for the treatment of HCV.     

Nigella sativa relieves the deleterious effects of ischemia reperfusion injury on liver

AIM： To determine whether Nigella sativa prevents hepatic ischemia-reperfusion injury to the liver. METHODS： Thirty rats were divided into three groups as sham （Group 1）, control （Group 2）, and Nigella sativa （NS） treatment group （Group 3）. All rats underwent hepatic ischemia for 45 min followed by 60 min period of reperfusion. Rats were intraperitoneally infused with only 0.9% saline solution in group 2. Rats in group 3 received NS （0.2 mL/kg） intraperitoneally, before ischemia and before reperfusion. Blood samples and liver tissues were harvested from the rats, and then the rats were sacrifi ced. Serum aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, and lactate dehydrogenase （LDH） levels were determined. Total antioxidant capacity （TAC）, catalase （CAT）, total oxidative status （TOS）, oxidative stress index （OSI） and my-eloperoxidase （MPO） in hepatic tissue were measured. Also liver tissue histopathology was evaluated by light microscopy. RESULTS： The levels of liver enzymes in group 3 weresignifi cantly lower than those in the group 2. TAC in liver tissue was significantly higher in group 3 than in group 2. TOS, OSI and MPO in hepatic tissue were signifi cantly lower in group 3 than the group 2. Histological tissue damage was milder in the NS treatment group than that in the control group. CONCLUSION： Our results suggest that Nigella sativa treatment protects the rat liver against to hepatic ischemia-reperfusion injury.     

Optimizing hepatitis C therapy in HIV/hepatitis C virus (HCV) coinfected patients: Analysis of HCV viral kinetics on treatment

INTRODUCTION:

Hepatitis C virus (HCV) infection is potentially curable, but the sustained virological response (SVR) has been shown to be lower in patients coinfected HIV. A single-centre experience treating individuals with HCV and HIV coinfection is reported.

METHODS:

Twenty-one patients who received standard doses of pegylated interferon with weight-based dosing of ribavirin (mean 14.3 mg/kg) were retrospectively reviewed. Qualitative HCV polymerase chain reaction (PCR) was performed prospectively every four weeks if the patient remained HCV PCR positive. All patients with HCV genotype 1 were treated for 48 weeks. Patients with genotype 2 or 3 were treated for 24 weeks and 32 weeks to 36 weeks if their HCV RNA level was undetectable after four weeks (RVR4) or eight weeks (RVR8) of therapy, respectively. If RVR8 was not achieved, the treatment was continued for 48 weeks.

RESULTS:

There were no dropouts or dose reductions within the first 12 weeks of treatment. SVR status was available for 20 patients and adequate serum for viral kinetics analyses was available for 17 patients. Eighty per cent of the patients achieved SVR (50% genotype 1; 100% genotypes 2 and 3). The week 8 viral load remained elevated for all genotype 1 nonresponders.

DISCUSSION:

High effectiveness rates were seen, particularly in patients with HCV genotype 2 and 3 who were treated for shorter durations. HCV viral loads after eight weeks of therapy helped distinguish patients with HCV genotype 1 who would respond to therapy.     

丙型肝炎病毒感染者血清核心蛋白水平与抗病毒治疗效果相关性分析

目的 探讨血清HCV核心蛋白与抗病毒治疗效果的相关性.方法 采用ELISA法检测162例丙型肝炎患者血清HCV核心蛋白量,采用荧光定量PCR检测HCV RNA;观察35例接受聚乙二醇干扰素+利巴韦林治疗的患者在治疗前、治疗4周、治疗24周、治疗48周时HCV RNA与HCV核心蛋白水平,比较两者血清水平随临床疗效变化的一致性.两组计量资料比较采用t检验,多组配对资料进行方差分析.结果 162例患者中56例HCV RNA≤1×103IU/mL,HCV核心蛋白吸光度(A)值为0.100±0.029;9例1×103IU/mL＜HCV RNA≤1×104 IU/mL,A值为0.246±0.213;11例1×104IU/mL＜HCV RNA≤1×105IU/mL,A值为0.235±0.179;28例1×105 IU/mL＜HCV RNA≤1×106IU/mL,A值为0.422±0.319;51例1×106IU/mL＜HCVRNA≤1×107IU/mL,A值为0.603±0.330;7例1×107IU/mL＜HCV RNA≤1×108IU/mL,A值为0.900±0.379.对HCV RNA与HCV核心蛋白水平进行相关性分析,结果呈正相关(r=0.36,P＜0.05).35例接受抗病毒治疗患者治疗前、治疗4周、治疗24周和治疗48周时的核心蛋白A值分别为0.564±0.296、0.144±0.062、0.091±0.035和0.112±0.103.与抗病毒治疗前比较,治疗4周、24周、48周患者血清HCV核心蛋白水平均明显下降(t=8.563、9.195、9.250,均P＜0.05);治疗4周与24周比较,差异也有统计学意义(t=4.301,P＜0.05).结论 在疾病进程及抗病毒治疗过程中血清HCV核心蛋白的量的变化与HCV病毒载量变化是平行的.     

Chaperones in hepatitis C virus infection

The hepatitis C virus(HCV) infects approximately 3% of the world population or more than 185 million people worldwide. Each year, an estimated 350000-500000 deaths occur worldwide due to HCV-associated diseases including cirrhosis and hepatocellular carcinoma. HCV is the most common indication for liver transplantation in patients with cirrhosis worldwide. HCV is an enveloped RNA virus classified in the genus Hepacivirus in the Flaviviridae family. The HCV viral life cycle in a cell can be divided into six phases:(1) binding and internalization;(2) cytoplasmic release and uncoating;(3) viral polyprotein translation and processing;(4) RNA genome replication;(5) encapsidation(packaging) and assembly; and(6) virus morphogenesis(maturation) and secretion. Many host factors are involved in the HCV life cycle. Chaperones are an important group of host cytoprotective molecules that coordinate numerous cellular processes including protein folding, multimeric protein assembly, protein trafficking, and protein degradation. All phases of the viral life cycle require chaperone activity and the interaction of viral proteins with chaperones. This review will present our current knowledge and understanding of the role of chaperones in the HCV life cycle. Analysis of chaperones in HCV infection will provide further insights into viral/host interactions and potential therapeutic targets for both HCV and other viruses.     

Emerging clinical and therapeutic applications of Nigella sativa in gastroenterology

Nigella sativa decreases DNA damage and thereby prevents initiation of carcinogenesis in colonic tissue secondary to exposure to toxic agents such as azoxymethane. N. sativa is of immense therapeutic benefit in diabetic individuals and those with glucose intolerance as it accentuates glucose-induced secretion of insulin besides having a negative impact on glucose absorption from the intestinal mucosa. N. sativa administration protects hepatic tissue from deleterious effects of toxic metals such as lead, and attenuates hepatic lipid peroxidation following exposure to chemicals such as carbon tetrachloride.     

APRI as a predictor of early viral response in chronic hepatitis C patients

AIM: To evaluate the aspartate aminotransferase (AST) to platelet ratio index (APRI) as a predictive factor of early viral response in chronic hepatitis C naive patients.METHODS: We performed an ambispective casecontrol study. We enrolled chronic hepatitis C naive patients who were evaluated to start therapy with PEGylated interferon a-2b (1.5 mg/kg per week) and ribavirin (> 75 kg: 1200 mg and < 75 kg: 1000 mg). Patients were allocated into two groups, group 1: Hepatitis C patients with early viral response (EVR), group 2: Patients without EVR. Odds ratio (OR) and 95% confidence interval (CI) were calculated to assess the relationship between each risk factor and the EVR in both groups.RESULTS: During the study, 80 patients were analyzed, 45 retrospectively and 35 prospectively. The mean ± SD age of our subjects was 42.9 ± 12 years; weight 70 kg (± 11.19), AST 64.6 IU/mL (± 48.74), alanine aminotransferase (ALT) 76.3 IU/mL (± 63.08) and platelets 209 000 mill/mm3 (± 84 429). Fifty-five (68.8%) were genotype 1 and 25 (31.3%) were genotype 2 or 3; the mean hepatitis C virus RNA viral load was 2 269 061 IU/mL (± 7 220 266). In the univariate analysis, APRI was not associated with EVR [OR 0.61 (95% CI 0.229-1.655, P = 0.33)], and the absence of EVR was only associated with genotype 1 [OR 0.28 (95% CI 0.08-0.94, P = 0.034)]. After adjustment in a logistic regression model, genotype 1 remains significant.CONCLUSION: APRI was not a predictor of EVR in chronic hepatitis C; Genotype 1 was the only predictive factor associated with the absence of EVR in our patients.     

PNPLA3 polymorphisms are associated with raised alanine aminotransferase levels in hepatitis C virus genotype 3

Background and study aimsHepatitis C virus (HCV) infection is one of the leading causes of end-stage liver diseases. This study aimed to determine the association between polymorphisms in interleukin 28B (IL28B), PNPLA3, toll-like receptor 7 (TLR7), nucleotide-binding oligomerization domain-containing protein 2 (NOD2) and retinoic inducible gene-I (RIG-I) and HCV genotype and clinical presentation in an Indian population.Patients and methodsA total of 500 patients with chronic HCV were enrolled in 19 centres across India. Genomic DNA was extracted from whole blood samples, and single nucleotide polymorphisms (SNPs) for IL28B, PNPLA3, TLR7, NOD2 and RIG-I genes were genotyped by real-time PCR using a TaqManSNP genotyping assay.ResultsThe mean age of the patients was 45 + 13 years, and the most common genotype observed was HCV genotype 3 (54%), followed by genotype 1 (24%). Although the allelic frequencies of TLR7, NOD2 and RIG-I were in significant disequilibrium in HCV patients compared with those in controls, the PNPLA3 polymorphism correlated significantly with higher viral load and alanine aminotransferase (ALT) levels in genotype 3 patients. Patients with PNPLA3 CG/GG genotypes, along with IL28B genotype CC, had higher levels of ALT than those with other genotypes.ConclusionThese results indicate that PNPLA3 polymorphisms are associated with higher ALT levels in HCV genotype 3 patients in India and can help in identifying people who are at greater risk of developing HCV-associated liver diseases.     

乙型肝炎病毒X基因-丙型肝炎病毒C基因共表达对血管内皮细胞生长因子的影响

目的建立乙型肝炎病毒X基因-丙型肝炎病毒C基因（HBV X—HCV C）共表达HepG2细胞模型，并探讨其对血管内皮细胞生长因子表达的影响。方法双酶切质粒pXT1—X，得到完整的HBV X基因片段后，将其插入到质粒PBK—CMV和PBK—HCV C的相应酶切位点，得到重组质粒PBK—X和PBK—X—C；再将质粒PBK—CMV、PBK—X、PBK—HCV C和PBK—X—C分别导入肝癌细胞株HepG2中，G418筛选，逆转录聚合酶链反应、Western blot鉴定HBV X和HCV C蛋白表达，免疫组织化学、Western blot检测血管内皮细胞生长因子蛋白质表达。结果质粒PBK—CMV、PBK—X、PBK—HCV C和PBK—X—C在HepG2细胞中有稳定表达。共表达HBV X—HCV C蛋白的细胞血管内皮细胞生长因子蛋白质表达较转染空载体的细胞及单独表达HBV X、HCV C蛋白的细胞明显升高。结论HBV X—HCV C共表达能显著上调血管内皮细胞生长因子蛋白质表达，提示HBV、HCV可能具有协同致癌作用。     

Evidence for a relation between the viral load and genotype and hepatitis C virus-specific T cell responses

BACKGROUND/AIMS: The reason why patients with hepatitis C virus (HCV) genotype non-1 infection respond better to antiviral therapy than patients with genotype 1 infection is not known. The aim of this study is to explore the relation between the viral genotype, viral load, and the endogenous T cell response. METHODS: The viral genotype, the viral load, and the endogenous proliferative T cell response to the non-structural 3 protein (NS3) was analysed using serum and peripheral blood mononuclear cells from 103 patients with chronic HCV infection. RESULTS: Among 71 nontreated patients a T cell response was more common among those infected by genotype 3, as compared to those infected with genotype 1 (P<0.05). Among 32 patients undergoing antiviral therapy, presence of a T cell response was more common in genotype non-1 infected patients than in those infected by genotype 1 (P<0.01). Presence of a T cell response was related to a more rapid viral clearance (P<0,05), a negative HCV RNA test at week 12 (P<0.05), and a shorter viral half-life (P<0.05). CONCLUSIONS: The presence of an NS3-specific T cell response is related to the viral genotype and to a more rapid clearance of HCV RNA during antiviral therapy.     

HBV X基因-HCV C基因cDNA融合基因真核表达载体的构建及其表达

目的：构建HBV X-HCV C融合基因真核表达载体，并获得稳定表达该基因的HepG2细胞株。方法：双酶切质粒pXT1-X，得到完整的HBV X基因片段后，将其插入到质粒PBK-CMV和PBK-HCVC的相应酶切位点，得到重组质粒PBK-X和PBK-X-C；再将质粒RBK-CMV、PBK-X、PBK-HCV C和PBK-X-C分别导入肝癌细胞株HepG2中，G418筛选，RT-PCR、蛋白印迹鉴定HBV X和HCV C蛋白表达。结果：质粒PBK-CMV、PBK-X、PBK-HCV C和PBK-X-C在HepG2细胞中有稳定表达。结论：成功构建HBV X-HCVC融合基因真核表达载体，并获得稳定表达该基因的HepG2细胞株。     

合并乙型肝炎病毒感染对丙型肝炎病毒核心抗原检测的影响

目的 探讨合并HBV感染对慢性HCV感染者血清丙型肝炎病毒核心抗原(HCVcAg)检出情况的影响. 方法 收集2005年12月-2009年10月慢性丙型肝炎患者和HBV/HCV合并感染者资料,检测血清HCVcAg和HCV RNA,对后者血清进行HBV DNA、HBeAg检测,分析HCVcAg检出率与HBeAg、HBV DNA定量检测的关系.用独立两组多分类的X2检验方法进行统计学分析. 结果 共收集88例慢性丙型肝炎患者和62例HBV/HCV合并感染者资料,血清HCVcAg的检出率分别为72.7％(64/88)和38.7％ (24/62),两者比较,x2= 17.358,P＜0.01,差异有统计学意义.HCV RNA检出率分别为81.8％ (72/88)和53.2％ (33/62),两者比较,x2=20.110,P＜0.01,差异有统计学意义.62例HBV/HCV合并感染者血清中,HBeAg阳性和HBeAg阴性感染者HCVcAg检出率分别为28.6％ (12/42)和60.0％ (12/20),两者比较,x2=5.641,P=0.011,差异有统计学意义.HCV RNA阳性率分别为42.9％ (18/42)和80.0％ (16/20),两者比较,X2=7.547,P＜ 0.01,差异有统计学意义.HBV DNA阳性和阴性时HCVcAg检出率分别为39.1％ (18/46)和37.5％ (6/16),两者比较,P＞0.05,差异无统计学意义.与单纯HCV感染者血清HCVcAg检出率72.7％ (64/88)比较,HBeAg阴性合并感染者为60.0％ (12/20),x2=1.266,P=0.261,差异无统计学意义;HBV DNA阴性合并感染者为37.5％ (6/16),x2=7.635,P＜0.01,差异有统计学意义.结论 HBV/HCV合并感染时HCVcAg检出率较低,可能是由于HBeAg抑制HCV的复制,从而减少HCVcAg的表达所致.     

丙型肝炎病毒核心蛋白DNA结合区的测定

目的 阐明丙型肝炎病毒（HCV）核心蛋白的DNA结合特性及其意义。方法 在大肠杆菌中表达谷胱甘肽转移酶（GST）融合HCV核心蛋白不同长度片段,并进行纯化。经聚丙烯酰胺凝胶电泳后,通过更换缓冲液的方法将这些蛋白片段相对固定在凝胶中,用γ－^32P[4667-0091]ATP标记人工合成的DNA进行2次电泳,通过放射自影检测核心蛋白的DNA结合区。结果 HCV核心蛋白N端第10－16位氨基酸残基和第46－70位氨基酸残基可独立地结合DNA,核心蛋白既能和单链DNA结合又能和双链DNA结合,对单双链DNA的结合域相同。对所结合的DNA序列无选择性。结论 HCV核心蛋白的N端含有两个DNA结合区,结合区序列与核内转移信号的部分重迭以及对结合靶DNA序列的非选择性可能是HCV核心蛋白具有多功能的基础。     

Neutralizing antibodies in hepatitis C virus infection

Hepatitis C virus (HCV) is a major cause of hepatitis world-wide. The majority of infected individuals develop chronic hepatitis which can then progress to liver cirrhosis and hepatocellular carcinoma. Spontaneous viral clearance occurs in about 20%-30% of acutely infected individuals and results in resolution of infection without sequaelae. Both viral and host factors appear to play an important role for resolution of acute infection. A large body of evidence suggests that a strong, multispecific and long-lasting cellular immune response appears to be important for control of viral infection in acute hepatitis C. Due too the lack of convenient neutralization assays, the impact of neutralizing responses for control of viral infection had been less defined. In recent years, the development of robust tissue culture model systems for HCV entry and infection has finally allowed study of antibody-mediated neutralization and to gain further insights into viral targets of host neutralizing responses. In addition, detailed analysis of antibody-mediated neutralization in individual patients as well as cohorts with well defined viral isolates has enabled the study of neutralizing responses in the course of HCV infection and characterization of the impact of neutralizing antibodies for control of viral infection. This review will summarize recent progress in the understanding of the molecular mechanisms of antibody-mediated neutralization and its impact for HCV pathogenesis.     

Status of hepatitis C virus vaccination:Recent update

Hepatitis C virus(HCV) infection is still a major public health problem worldwide since its first identification in 1989. At the start, HCV infection was post-transfusion viral infection, particularly in developing countries. Recently, due to iv drug abuse, HCV infection became number one health problem in well-developed countries as well. Following acute HCV infection, the innateimmune response is triggered in the form of activated coordinated interaction of NK cells, dendritic cells and interferon α. The acquired immune response is then developed in the form of the antibody-mediated immune response(ABIR) and the cell-mediated immune response(CMIR). Both are responsible for clearance of HCV infection in about 15% of infected patients. However, HCV has several mechanisms to evade these antivirus immune reactions. The current review gives an overview of HCV structure, immune response and viral evasion mechanisms. It also evaluates the available preventive and therapeutic vaccines that induce innate, ABIR, CMIR. Moreover, this review highlights the progress in recent HCV vaccination studies either in preclinical or clinical phases. The unsatisfactory identification of HCV infection by the current screening system and the limitations of currently available treatments, including the ineligibility of some chronic HCV patients to such antiviral agents, mandate the development of an effective HCV vaccine.     

Occult hepatitis B virus infection in Egypt

The emerging evidence of the potentially clinical importance of occult hepatitis B virus (HBV) infection (OBI) increases the interest in this topic. OBI may impact in several clinical contexts, which include the possible transmission of the infection, the contribution to liver disease progression, the development of hepatocellular carcinoma, and the risk of reactivation. There are several articles that have published on OBI in Egyptian populations. A review of MEDLINE database was undertaken for relevant articles to clarify the epidemiology of OBI in Egypt. HBV genotype D is the only detectable genotype among Egyptian OBI patients. Higher rates of OBI reported among Egyptian chronic HCV, hemodialysis, children with malignant disorders, and cryptogenic liver disease patients. There is an evidence of OBI reactivation after treatment with chemotherapy. The available data suggested that screening for OBI must be a routine practice in these groups of patients. Further studies needed for better understand of the epidemiology of OBI among Egyptian young generations after the era of hepatitis B vaccination.