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<p><b>OBJECTIVE</b>To study the resistant rate of hepatitis B virus (HBV) to ADV and the dynamic evolution of HBV in lamivudine (Lam)-resistant chronic hepatitis B (CHB) patients.</p><p><b>METHODS</b>Twenty-three Lam-resistant CHB patients were assigned to a 10mg/d ADV monotherapy for 68-116 weeks. The baseline and different time point blood samples after ADV monotherapy were analyzed for ADV-resistant mutations using direct sequencing of PCR products; the evolution of HBV mutations was examined by clonal analysis of serial samples from one patient infected with ADV-associated resistant HBV strains.</p><p><b>RESULTS</b>The cumulative incidence of genotypic ADV resistance at weeks 48 and 96 was 4.3% and 10.5% respectively respectively. The evolution analysis of HBV mutant strains in an ADV-resistant CHB patient showed that the proportion of YMDD mutants gradually decreased with rtA181S mutants increasing over time after ADV monotherapy, and that rtA181S+N236T mutants became the predominant strains during prolonged ADV monotherapy. The addition of Lam to the ongoing ADV treatment had poorer antiviral response in the patient with rtA181S or rtA181S+N236T mutant infection; one clone with multi-drug resistant mutations was selected during Lam and ADV combination therapy.</p><p><b>CONCLUSION</b>Increased risk of adefovir resistance and selection of multi-drug resistant mutations are associated with long-term ADV monotherapy in patients with Lam-resistant chronic hepatitis B.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Adenine , Therapeutic Uses , Antiviral Agents , Therapeutic Uses , Drug Resistance, Viral , Evolution, Molecular , Hepatitis B virus , Classification , Genetics , Hepatitis B, Chronic , Drug Therapy , Virology , Lamivudine , Pharmacology , Organophosphonates , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the phenotypes and functions of peripheral blood monocyte derived dendritic cells (DC) of chronic hepatitis B (CHB) patients with different HBV DNA loads.</p><p><b>METHODS</b>Twenty-eight CHB patients were included in this study. All patients were treated with nucleoside analogues (lamivudine or LdT or adefovir) for 24 weeks. Peripheral blood HBV DNA loads and liver biopsies were assessed before and after the treatment. The patients were divided into two groups according to their peripheral blood HBV DNA loads: a high-load group with HBV DNA loads higher than 10(5) copies/ml, and a low-load group with HBV DNA loads lower than 10(3) copies/ml. Ten healthy people were included as controls. Peripheral blood DC of each subject was enriched. The phenotypes of DC were subjected to flow cytometric analysis. The lymphocyte allo-stimulatory capacity of DC was evaluated through MTT assay. IL-10 and IL-12 production were quantified by ELISA.</p><p><b>RESULTS</b>DC proliferated successfully when stimulated by cytokines in vitro; however, DC of the CHB patients proliferated much slower than those of the healthy controls. The expression of DC surface molecules such as HLA-DR, CD86, CD80 and CD83 had a positive rate of over 80% in the normal population. However in our CHB patients they showed lower than normal expressions, especially the HLA-DR, CD86, CD80 and CD83, but the differences were not significant between the two groups with different virus loads. The stimulatory capacity of the DC in mixed lymphocyte reaction showed no difference between the two groups of patients, but both were lower than that of the healthy controls. The production of IL-12 and IL-10 also decreased significantly in the patients.</p><p><b>CONCLUSIONS</b>Peripheral DC of CHB patients have some defects in their phenotypes and their stimulatory capacity. The changes in phenotypes and down-regulation of the functions are not relevant to peripheral HBV DNA loads of the patients.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Dendritic Cells , Allergy and Immunology , Metabolism , Hepatitis B virus , Genetics , Hepatitis B, Chronic , Blood , Allergy and Immunology , Virology , Phenotype , Viral LoadABSTRACT
Illegally paid blood donation was a risk factor for HIV acquisition exclusively in Henan and Hubei Provinces of China, and not in Shanghai. Nucleotide sequences in the gag and env genes of HIV-1 were compared between isolates from Henan and Shanghai regions of China to test whether an expected higher degree of a common source of infections from this unique blood donation transmission risk would be evident as decreased variation among Henan isolates in an exploratory cross-sectional analysis. Among 38 isolates studied, 23 of 23 (100%) from Henan and 8 of 15 (54%) from Shanghai were subtype B. In addition, fewer sequence differences were found in gp41 of subtype B isolates from Henan than from Shanghai isolates. Further studies with additional controls are therefore warranted to confirm the role of the degree of a common source of infections in differences in HIV variation across populations.
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Objective To establish a convenient,accurate and practical method for detection of adefovir dipivoxil resistance-as- sociated mutation in hepatitis B virus:rtA181V/T/S and rtN236T mutations.Methods According to HBV complete sequences in GenBank,two pairs of primers were designed to amplify the region of HBV reverse transcriptase in order to introduce a BglI restriction site upon PCR product of wild type (wt) and a BseDI restriction site upon PCR product of rt236 mutant type.After amplification,the PCR products were digested with BglI and BseDI separately.We used this method to detect wild,rt181 mu- tant,rt236 mutant plasmids and 3 chronic hepatitis B patients' serum with obvious ADV resistance-associated mutations.We also tested the sensitivity of this method by mixing the wild and mutant plasmids in different proportions.Results The method could detect rt181 and rt236 mutations simultaneously.The result of RFLP analysis was in accordance with that of DNA se- quencing and cloning analysis.This method could detect the mutants even when they comprised only 10% of the total virus population.Conclusions The PCR-RFLP method with high sensitivity can detect rt181 and rt236 mutations simultaneously.It can be used for early detection of ADV resistance-associated mutation in hepatitis B virus.
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Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study,and ten healthy persons were recruited as controls.The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granuloeyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation.Then the phenotype of DCs was identified by micro- scope.The expressions of the phenotypes[histocompatibility leukocyte antigen(HLA)-DR,CD80, CD86,CD83]of immature and mature DCs were measured by flow cytometer.Furthermore,the ex- pression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively.Results As for healthy volunteers,the expressions of CD80, CD86,HLA-DR and CD83 on DCs at the 7th day,which were(82.35?8.67)%,(79.61?10.08)%, (92.79?8.48)% and (83.76?5.47)% respectively,were significantly higher than those at the 5th day which were(28.31?8.79) %,(31.17?11.23)%,(27.61?10.28)% and (23.46?11.53)% respec- tively(P0.05).The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P0.05).And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P
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Objectives To investigate the clinical features,prognosis and risk factors of patients with cryptococcal meningitis.Methods Totally 146 patients with cryptococcal meningitis who were hospitalized in Huashan Hospital from January 2000 to December 2006 were enrolled in this study.The clinical data including diagnosis and misdiagnosis,experimental and etiology tests,treat- ments and prognosis from all the patients were analyzed retrospectively.Results Among the 146 patients enrolled in the study,78 patients(53.4%)had concomitant diseases.The misdiagnosis rate of all patients was 72.6%(106/146).The positive rate of cerebrospinal fluid(CSF)India ink smear was 59.6%(87/106),while 43.2%(63/146)cases of cryptococcus neoformans culture in CSF was positive.The positive rate of Latex agglutination test(LAT)was 91.7%(134/146)in CSF among all patients.The treatments were as follows:combination of Amphotericin B(AmpB)or its lipid formula- tions with flucytosine(5-FC)(98 cases),including combination with Fluconazole initally(62 cases), single therapy of Fluconazole(13 cases).Ommaya implanted for lateral cerebral ventricle drainage(53 cases)and AmpB intrathecal injection(53 cases).The average dose of AmpB is 3.06 g.The course of treatment lasted from 12 weeks to 20 months.There were 104 patients(71.2%)cured,27(18.5%) improved,15(10.3%)died and 34(23.3%)relapsed.Conclusions High misdiagnosis rate is common in patients with cryptococcal meningitis.Immunodeficiency is the major risk factor for cryp- tococcal meningitis.CSF LAT is the most sensitive diagnostic test.Early diagnosis,combination of AmpB with 5-FC antifungal therapy and control of acute intracranial hypertension are the keys to im- prove prognosis of cryptococcal meningitis.