高活性抗逆病毒疗法减少HIV病晚期患者的死亡率和发病率

在采用高活性抗逆病毒治疗(HAART)的HIV感染患者中,与艾滋病有关的死亡率和发病率下降,但以前的研究也许是因为治疗上的其他改变而使人迷惑不解。为了评价HAART对晚期艾滋病与贫血患者的疗效,美国的Murphy等研究人员进行了一项前瞻性多中心队列研究。

重庆黔江区艾滋病患者抗病毒治疗前后CD4＋T细胞水平分析

自国家开展免费抗病毒治疗以来，重庆市黔江区于2006年开始艾滋病的免费治疗，到2011年累计共有20例患者接受了免费高活动性抗逆病毒治疗（HAART）。为了解这些患者抗病毒治疗CD4＋T淋巴细胞检测情况以及变化特点，作者对这些患者CD4＋T细胞淋巴结果进行分析统计，比较治疗前后CD4＋T细胞计数及不同治疗时段的CD4＋T淋巴细胞的变化特点，现报道如下。

在博茨瓦纳比较母乳喂养加婴儿预防性应用叠氮胸苷6个月与配方奶加婴儿叠氮胸苷1个月以减少母婴HIV传播的一项随机试验:Mashi研究

Context: Postnatal transmission of human immunodeficiency virus-1 (HIV) via breastfeeding reverses gains achieved by perinatal antiretroviral interventions. Objective: To compare the efficacy and safety of 2 infant feeding strategies for the prevention of postnatal mother-to-child HIV transmission. Design, Setting, and Patients: A 2×2 factorial randomized clinical trial with peripartum (single-dose nevirapine vs placebo) and postpartum infant feeding (formula vs breastfeeding with infant zidovudine prophylaxis) interventions. In Botswana between March 27, 2001, and October 29, 2003, 1200 HIV-positive pregnant women were randomized from 4 district hospitals. Infants were evaluated at birth, monthly until age 7 months, at age 9 months, then every third month through age 18 months. Intervention: All of the mothers received zidovudine 300 mg orally twice daily from 34 weeksgestation and during labor. Mothers and infants were randomized to receive single-dose nevirapine or placebo. Infants were randomized to 6 months of breastfeeding plus prophylactic infant zidovudine (breastfed plus zidovudine), or formula feeding plus 1 month of infant zidovudine (formula fed). Main Outcome Measures: Primary efficacy (HIV infection by age 7 months and HIV-free survival by age 18 months) and safety (occurrence of infant adverse events by 7 months of age) end points were evaluated in 1179 infants. Results: The 7-month HIV infection rates were 5.6%(32 infants in the formula-fed group) vs 9.0%(51 infants in the breastfed plus zidovudine group) (P=.04; 95%confidence interval for difference,-6.4%to-0.4%). Cumulative mortality or HIV infection rates at 18 months were 80 infants (13.9%, formula fed)-vs 86 infants (15.1%breastfed plus zidovudine) (P=.60; 95%confidence interval for difference,-5.3%to 2.9%). Cumulative infant mortality at 7 months was significantly higher for the formulafed group than for the breastfed plus zidovudine group (9.3%vs 4.9%; P=.003), but this difference diminished beyond month 7 such that the time-to-mortality distributions through age 18 months were not significantly different (P=.21). Conclusions: Breastfeeding with zidovudine prophylaxis was not as effective as formula feeding in preventing postnatal HIV transmission, but was associated with a lower mortality rate at 7 months. Both strategies had comparable HIV-free survival at 18 months. These results demonstrate the risk of formula feeding to infants in sub-Saharan Africa, and the need for studies of alternative strategies. Trial Registration: clinical trials. gov Identifier:

Impact of human immunodeficiency virus infection on the course of hepatitis C virus infection: A meta-analysis

AIM: To analyze the influence of human immunodeficiency virus (HIV) infection on the course of hepatitis C virus (HCV) infection. METHODS: We performed a meta-analysis to quantify the effect of HIV co-infection on progressive liver disease in patients with HCV infection. Published studies in the English or Chinese-language medical literature involving cohorts of HIV-negative and -positive patients coinfected with HCV were obtained by searching the PUBMED, EMBASE and CBM. Data were extracted independently from relevant studies by 2 investigators and used in a fixed-effect meta analysis to determine the difference in the course of HCV infection in the 2 groups. RESULTS: Twenty-nine trails involving 16 750 patients were identified including the outcome of histological fibrosis or cirrhosis or de-compensated liver disease or hepatocellular carcinoma or death. These studies yielded a combined adjusted odds ratio (OR) of 3.40 [95% confidence interval (CI) = 2.45 and 4.73]. Of note, studies that examined histological fibrosis/ cirrhosis, decompensated liver disease, hepatocellular carcinoma or death had a pooled OR of 1.47 (95% CI = 1.27 and 1.70), 5.45 (95% CI = 2.54 and 11.71), 0.76 (95% CI = 0.50 and 1.14), and 3.60 (95% CI = 3.12 and 4.15), respectively. CONCLUSION: Without highly active antiretroviral therapies (HAART), HIV accelerates HCV diseaseprogression, including death, histological fibrosis/ cirrhosis and decompensated liver disease. However, the rate of hepatocellular carcinoma is similar in persons who had HCV infection and were positive for HIV or negative for HIV.

Expression of Major Antigen Domains of E2 Gene of CSFV and Analysis of its Immunological Activity

E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this study, four antigen domains (ABCD) of the E2 gene was cloned from CSFV Shimen strain into the retroviral vector pBABE puro and expressed in eukaryotic cell (PK15) by an retroviral gene expression system, and the activity of recombinant E2 protein to induce immune responses was evaluated in rabbits. The results indicated that recombinant E2 protein can be recognized by fluorescence antibodies of CSFV and CSFV positive serum (Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China) using Western blot, indirect immunofluorescence antibody test (IFAT) and ELISA, Furthermore, anti-CSFV specific antibodies and lymphocyte proliferation were elicited and increased by recombinant protein after vaccination. In the challenge test, all of rabbits vaccinated with recombinant protein and Chinese vaccine strain (C-strain) were fully protected from a rabbit spleen virus challenge. These results indicated that a retroviral-based epitope-vaccine carrying the major antigen domains of E2 is able to induce high level of epitope-specific antibodies and exhibits similar protective capability with that induced by the C-strain, and encourages further work towards the development of a vaccine against CSFV infection.

Role of upper endoscopy in diagnosing opportunistic infections in human immunodeficiency virus-infected patients

Highly active antiretroviral therapy (HAART) has dramatically decreased opportunistic infections (OIs) in human immunodef iciency virus (HIV)-infected patients. However,gastrointestinal disease continues to account for a high proportion of presenting symptoms in these patients. Gastrointestinal symptoms in treated patients who respond to therapy are more likely to the result of drug-induced complications than OI. Endoscopic evaluation of the gastrointestinal tract remains a cornerstone of diagnosis,especially in patients with advanced immunodeficiency,who are at risk for OI. The peripheral blood CD4 lymphocyte count helps to predict the risk of an OI,with the highest risk seen in HIV-infected patients with low CD4 count (< 200 cells/mm3). This review provides an update of the role of endoscopy in diagnosing OI in the upper gastrointestinal tract in HIV-infected patients in the era of HAART.

An Assessment of the Level of Knowledge of HIV-Infected Patients about Highly Active Antireteroviral Therapy and Waiting Times and Their Influence on Antiretroviral Therapy Adherence at a Primary Healthcare Centre of South Africa

Objectives： The study assessed if the level of knowledge of HIV-infected about HAART and waiting-times in the PHC （primary healthcare） clinic have an influence on antiretroviral adherence. Methods： A descriptive-cross-sectional study was conducted in South Africa. Data collected uses a standardized-questionnaire and face-to-face-exit interviews. Pill-count technique was performed and a value of≥ 95% acceptable. Data were analysed using SPSS. Univariate-factors associated with poor-adherence to knowledge about HAART and waiting times were assessed using ANOVA and p ≤ 0.05 considered statistically significant. Key findings： Of 86 enrolled, 63（73.3%） were females and 23（26.7%） males, with mean-age （± SD） of 35.6（±9.6） years and on HAART for 35.5（± 31.8） months ranging from 1-137. Of these, 27（31.40%） and 25（29.07%） were on WHO stages 2 and 3 respectively. Adherence-rates computed from 32 patients, 23（71.9%） revealed poor adherence-rates. The level of knowledge about HAART in terms of names of tablets, correct-dose, frequency, adverse-effects had no influence on ARV-adherence （p _〉 0.05）. Of 23 non-compliant, 10 （40%） gave the reason of drugs-unavailability, 7（30%） adverse-effects, 5（20%） drugs＇ complexity, and 1（10%） too busy to take them. Waiting areas associated with poor ARV-adherence were reception （p = 0.028）, doctors （p = 0.027）, while nurse＇s station （p = 0.29） and pharmacy （p = 0.43） revealed acceptable ARV-adherence.

Construction and expression of the bicistronic expression vector with RANTES and SDF-1 genes

Objective: To construct bicistronic expression vector with RANTES and SDF-1 genes, the ligands of HIV-1 principal coreceptors, and identify its expression. Methods: RANTES-KDEL was amplified from plasmid pCMV-R-K by PCR and cloned into eukaryotic expression vector pCMV-S/K. Gene transfection into HeLa cells was carried out by lipofectin. Indirect immumofluorescence and radioimmunoprecipitation were used to confirm the expression of RANTES and SDF-1. Results: The construction of pCMV-R-K-S-K was confirmed by enzymatic digestion and sequencing. RANTES and SDF-1 were shown expressed in HeLa cells by indirect immumofluorescence and radioimmunoprecipitation. Conclusion: pCMV-R-K-S-K was constructed and expressed in cell line Hela successfully, which will contribute to further study of gene therapy of AIDS by HIV-1 coreceptors knockout.

A Few Specialized Issues That Should Be Focused on Anti-HIV Drug Evaluation In Vitro

Since the introduction of antiretroviral therapy (ART), the lifespan and quality of life of patients infected with HIV have been significantly improved. But treatment efficacy was compromised eventually by the development of resistance to antiretroviral drugs, and more new anti-HIV drugs with lower toxicity and higher activity were needed. Based on the experience and lessons learned from the treatment in the developed countries, US FDA suggested that more pharmacodynamical researches should be considered ahead of the clinical trials. To facilitate the anti-HIV drug research and development, we reviewed a few specialized issues that should be focused on drug evaluations in vitro, including: 1) Mechanism of action studies, demonstrating the candidate drug's efficacy to specifically inhibit viral replication or a virus-specific function and confirm the drug target. 2) Drug resistance studies, selecting the drug-resistant variants in vitro and determining the activities inhibiting HIV isolates resistant to approved antiretroviral drugs of the same class. 3) Antiviral activity in vitro in the presence of serum proteins, ascertaining whether an investigational product is significantly bound by serum proteins. 4) Combination activity analysis, evaluating in vitro antiviral activity of an investigational product in two-drug combinations with other drugs approved.

Induction of APOBEC3G in human hepatoma cells by interferon-α stimulation

To clarify the role of APOBEC3G （A3G） in cellular defense against hepatitis B virus （HBV）, the expression of A3G in normal human liver and the regulation of the A3G expression in hepatoma cell line （HuH-7） were investigated. Expression level of APOBEC3s mRNA in human liver was determined by RT-PCR. HuH-7 and HepG2 cells were treated with various concentrations of IFN-α（0 U/ml, 100 U/ml, 500 U/ml, 1000 U/ml）for 12 h. The mRNA levels were measured by a quantitative RT-PCR, the results were normalized relative to the specimens without IFN-α stimulation. Total protein of HuH-7 cells treated with various concentrations of IFN-α for 48 h was subjected to Western blot analysis. For reporter gene assay, HuH-7 cells were transfected with the reporter plasmids containing IRF- E sites and its mutants with different lengths. Then the cells were treated with or without 1200 U/ml IFN-α for additional 12 h （ 1000 U/ml） after 24 h of transfection, and the cell lysate was prepared and assayed for lueiferase activity. It was found that normal human liver expressed the rnR_NA of A3G. A3G mRNA expression in HuH-7 and HepG2 cells were up-regulated by IFN-α stimulation in a dose-depen- dent manner. Western blot analysis indicated that A3G protein expression was also enhanced by IFN-α stimulation. Sequence analysis showed the existence of putative sites of IFN regulatory factor element （IRF-E） in 5＇ region of A3G gene upstream the initiation eodon. IFN-α stimulation results in 6- to 8- fold increase in lueiferase activity in cells transfeeted with the plasmid containing IRF-E sites of the 5＇ upstream sequences, whereas luciferase activity did not change in cells transfected with the plasmid containing mutant IRF-E sites or without IRF-E sites. As a conclusion, A3G are expressed in normal human liver. A3G expression was up-regulated by IFN-α stimulation in hepatoma cells and could be involved in host defense mechanisms against HBV. IRF-E site in 5＇ region of APOBEC3G gene upstream the initiation codon plays an important role in this process.

Antiretroviral Therapy through Barriers： A Prominent Role for Nanotechnology in HIV-1 Eradication from Sanctuaries

Abstract： In HIV-1 management, eradication of the virus from sanctuaries represents a major and challenging goal. The genital tract, gut associated lymphoid tissue, lymph nodes, central nervous system, macrophages and latently infected CD4＋ T lymphocytes are typical sites where H1V-1 compartmentalizes. To circumvent this problem, a consistent number of studies have focused on improving ARVs （antiretroviral drugs） delivery into sanctuary sites and different nanoteehnological approaches have been developed. Cellular HIV-1 sanctuaries （i.e. macrophages） can be reached by nanoformulation of ARVs or by activation of latently infected cells. Anatomical sanctuaries （i.e. brain or male genital tract） can be addressed by increasing the permeation of ARVs across tissue barriers, such as the blood-brain barrier or the blood-testis barrier, while ARVs concentration in lymph nodes can be enhanced by drug encapsulation in CD4-targeted nanoparticles.

Effect of Maternal Antibodies on the Pathogenesis of Avian Reovirus Infections in Broiler Chickens Using Real-Time Reverse Transcriptase Polymerase Chain Reaction

The effect of maternal antibodies on the pathogenesis of avian reovirus （ARV） was studied in commercial and specific pathogen free broilers （SPF） using a real-time reverse transcriptase （RT）-polymerase chain reaction （PCR） assay, along with the incidence and severity of morbidity, mortality, and gross lesions. ARV RNA was detected in cloacal swabs in both bird groups from the first day throughout the 21 days experiment. Commercial broiler chickens, which had high maternal antibodies against ARV, showed minimum clinical signs, gross lesions, and lower numbers of birds with viral RNA excretion, whereas specific pathogen free （SPF） broiler chickens, which did not have antibody against ARVs, had 30% mortality, more severe signs, and higher numbers of birds excreting viral RNA. The highest peak of SPF birds excreting viral RNA occurred during the time of maximum mortality. The protective effect of maternal antibody on ARV pathogenesis in broiler chickens correlated with the detection of ARV RNA using the real-time RT-PCR.

Tumor cell-specific gene transfer with retroviral vectors displaying single-chain antibody

OBJECTIVE: To specifically deliver the therapeutic gene to cancer cells and construct target retroviral vectors by inserting the single-chain variable antibody fragment into the retroviral envelope. METHODS: Single-chain antibody expression vector pET -20bScfv was constructed. Binding activity of the genetically engineered single-chain variable antibody fragment was verified by enzyme-linked immunosorbent assay (ELISA) and Western blot. At the same time, by means of polymerase chain reaction (PCR)-directed mutagenesis, the appropriate cloning site EcoT22/Sal was generated at the N-terminus of receptor-binding SU domain in the MoMLV env polypeptide. Then the single- chain antibody gene, encoding a functional antibody, was inserted into the cloning site. The Scfv-env fusion gene fragment was subcloned into CMV expression vector. The Lac-Z retrovirus that co-displayed the Scfv-env chimeric protein and wild-type env protein was produced by transfection of Psi 2 cells with retroviral plasmid and the fusion gene expressing plasmid.To confirm the specificity of the recombinant retrovirus, infection assays and competitive inhibition assays were performed. RESULTS: The results of ELISA and Western blot showed that the genetically engineered single-chain variable antibody fragment could bind to the SHG44 cells surface membrane antigen. Virus-binding assay, viral infection and competitive inhibition assays confirmed that the harvested virus efficiently bound to and infected SHG44 cancer cells expressing the relative membrane antigen specially via the recognition of the target antigen. CONCLUSION: These results imply that insertion of Scfv into the retroviral envelope can specifically deliver the interested gene into specific antigen-producing cancer cells.

国外新近批准主要新药（三十一）

欧盟批准依曲韦林用于合并疗法治疗业经其它抗逆病毒药物治疗失败之艾滋病毒-1感染患者 Johnson＆Johnson公司2008年8月29日宣布，欧盟委员会已批准其提交的抗艾滋病新药依曲韦林片剂（etravirine／Intelence）的新药申请，用于合用增效蛋白酶抑制剂和其它抗逆病毒药物治疗业经在先抗逆病毒疗法治疗且具病毒复制和病毒株业对一种非核苷类逆转录酶抑制剂（NNRTI）和其它抗逆病毒药物耐药证据之成人艾滋病毒-1感染患者。依曲韦林为Tibotec制药公司发现化合物，而Tibotec制药公司则是Johnson＆Johnson公司的子公司之一。依曲韦林先前业于今年早些时候在美、加、俄、韩、瑞士和阿根廷等国获得批准。