OBJECTIVETo investigate the relationship between the degree of week 24 HBV suppression and week 48 therapeutic response in entecavir-treated chronic hepatitis B patients in whom lamivudine treatment failed, so as to explore a useful predictor for efficacy of enticavir treatment.

METHODSThirty-three patients with chronic hepatitis B refractory to lamivudine were enrolled to receive treatment with entecavir 1.0 mg once daily. The patients were divided into 4 groups according to serum HBV DNA levels (copies/mL) at week 24: PCR-undetectable (less than 300 copies/ml); QL- less than 3 log10 copies/ml; 3 log10(-4) log10 copies/ml; greater than 4 log10 copies/mL, and the efficacy achieved at week 48 was evaluated.

RESULTSAt week 48, mean reductions of serum HBV DNA from baseline was 4.91 log10. HBV DNA became undetectable by PCR assay in 33.3 percent patients and ALT became normal in 75.8%. The lower the HBV DNA level achieved at week 24, the higher the proportion of patients in whom HBV DNA became undetectable by PCR and ALT normalization were acquired at week 48, and viral breakthrough at week 48 also decreased.

CONCLUSIONUndetectable HBV DNA by PCR at week 24 in entecavir-treated chronic hepatitis B patients who were refractory to lamivudine, suggests a better efficacy at week 48. The degree of week 24 suppression of HBV may be used as a predictor of long term outcome.

Adolescent ; Adult ; Aged ; Antiviral Agents ; pharmacology ; therapeutic use ; Drug Administration Schedule ; Drug Evaluation ; Guanine ; administration & dosage ; analogs & derivatives ; pharmacology ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Treatment Failure ; Young Adult

Acquired Immunodeficiency Syndrome ; prevention & control ; China ; Humans ; Infectious Disease Medicine ; methods ; trends

OBJECTIVETo investigate in vitro and in vivo anti-human herpes simplex virus effects of photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA).

METHODSGuinea pigs model of cutaneous herpes virus infection was applied, and Vero cells infected by HSV-I and HSV-II were used as experimental systems to observe the antiherpes effect of ALA-PDT.

RESULTSThe in vitro experiments showed that ALA-PDT has antiherpes effect on HSV-I and HSV-II, its effect was similar to that of acyclovir. The results of animal experiments showed that ALA-PDT had significant therapeutic effect on guinea pigs model of cutaneous herpes virus infection, the effect was dose-related.

CONCLUSIONALA-PDT could be effective in treating HSV infections, which may provide a new approach to the treatment of viral infections.

Aminolevulinic Acid ; therapeutic use ; Animals ; Cercopithecus aethiops ; Disease Models, Animal ; Female ; Guinea Pigs ; Herpes Simplex ; drug therapy ; virology ; Male ; Photochemotherapy ; Random Allocation ; Simplexvirus ; drug effects ; Skin Diseases, Viral ; drug therapy ; Treatment Outcome ; Vero Cells

OBJECTIVETo construct and express anti-human RBC and HIVgp160 fusion protein for rapid detection of antibody to HIV.

METHODSThe gene of the anti human RBC ScFv and HIV antigen were constructed together into expression vector. The fusion protein was expressed in E. coli.

RESULTSThe fusion protein was proved to be able to bind both anti-RBC and HIVgp160. It could cause agglutination of human RBC when HIVgp160 was present.

CONCLUSIONThe fusion protein has the potential in rapid detection of HIV.

Antibodies, Monoclonal ; immunology ; isolation & purification ; Autoantibodies ; immunology ; isolation & purification ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Erythrocytes ; immunology ; Gene Expression ; Genetic Vectors ; genetics ; HIV Antibodies ; blood ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; metabolism ; HIV Seropositivity ; blood ; Hemagglutination Tests ; methods ; Humans ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo establish an accurate and efficient method for detecting recent thymic output function and analyze the content of T-cell receptor (TCR) rearrangement excision circles (TRECs) within peripheral blood mononuclear cells (PBMCs).

METHODSAccording to the specific sequence of TCRdelta, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The standard quantitative template was constructed by T/A cloning. The method for detecting TRECs was established after optimization of reaction condition, then its specificity, sensitivity and stability were tested. Quantitative detection of TRECs in DNA of PBMCs from normal individuals and patients of chronic hepatitis B were preformed by real-time PCR using TaqMan technique.

RESULTSDetection of TRECs was quick and accurate by real-time fluorescence quantitative PCR. The CV value of Ct was 1.06%, the product was specific which was confirmed by electrophoresis and sequencing and the method showed high sensitivity. The mean value of TRECs from normal individuals was (7767.4 +/- 2369.5) copies/10(6)PBMCs in healthy controls at age 21.45 but (28,374.4 +/- 7820.4) copies/10(6)PBMCs in those at age 16.20 (P < 0.05). The mean value of TRECs from patients with chronic hepatitis B was (6480.9 +/- 2031.2) copies/10(6) PBMCs in those at age 21.45, which was statistically significant as compared with normal individuals at age 21.45.

CONCLUSIONReal-time fluorescence quantitative PCR for detecting the TRECs is an accurate, efficient and stable method and the recent thymic output function might decrease in patients with chronic hepatitis B.

Adult ; DNA Primers ; Female ; Gene Rearrangement, T-Lymphocyte ; Hepatitis B, Chronic ; blood ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; Reproducibility of Results ; Thymus Gland ; immunology ; metabolism ; Young Adult

OBJECTIVETo establish a mismatched polymerase chain reaction restricted fragment length polymorphism (mPCR-RFLP) method for detection of hepatitis B virus (HBV) mutation in precore A1896, and compare with direct sequencing for evaluating its applicability.

METHODSAccording to the principle of mPCR, 194bp gene fragments in HBV precore region was amplified. The products of PCR were digested by Bsu36I and subjected to agarose gel electrophoresis. A method for detecting procore A1896 mutation was established by restricted fragment length polymorphism. Totally 134 sera were analyzed by both mPCR-RFLP and direct sequencing methods. Two sera which were identified having mixed infection with precore wild and mutant strains by mPCR-RFLP also were analyzed by cloning and sequencing.

RESULTSFrom 134 sera, 117 could be analyzed for HBV precore 1896 situation by mPCR-RFLP method, 109 could be analyzed by sequencing. In 101 sera which could be analyzed by the two methods, 54 were mutant strains and 47 were wild strains. The results of both methods were completely compatible. There was no significant difference in detective rate of HBV precore A1896 mutation between the two methods. The sequences of five clones from one serum which was identified precore mutant by mPCR-RFLP were all A1896 mutant strains. Another serum was identified as mixed infection by mPCR-RFLP, one clone was A1896 mutant strain and four were G1896 wild strains. The results of mPCR-RFLP were verified by cloning.

CONCLUSIONCompared with sequencing, the mPCR-RFLP method is simple, accurate and can be used in large-scale surveys and clinical research.

Adult ; Aged ; DNA, Viral ; blood ; genetics ; isolation & purification ; Female ; Genetic Heterogeneity ; Hepatitis B ; blood ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Young Adult

OBJECTIVETo evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer.

METHODSAFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed.

RESULTSThe levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%.

CONCLUSIONDetection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Liver Cirrhosis ; blood ; diagnosis ; Liver Neoplasms ; blood ; diagnosis ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis

OBJECTIVETo observe the therapeutic effect of autologous cytokine-induced killer cells (CIK) on HBV DNA positive patients with liver cirrhosis.

METHODSHBV DNA positive 33 patients with cirrhosis were treated with CIK. Before and after cultured in vitro and post-treatment, CD3+, CD3+CD4+, CD3+CD8+, CD3+CD56+ cells, mDC and pDC were detected by flow cytometry. The indexes of virus and liver function were compared between pre- and post-treatment.

RESULTSCD3+, CD3+CD8+ cells and CD3+CD56+ cells were higher after cultured in vitro and after transfused back than those before culture (91.5 +/- 10.3, 74.4 +/- 9.9 vs. 67.9 +/- 12.8; 60.9 +/- 15.5, 37.3 +/- 15.1 vs. 27.9 +/- 10.9; 18.4 +/- 11.7, 14.5 +/- 7.5 vs. 10.6 +/- 7.1). The percentages of mDC and pDC also increased after-treatment vs. pre-treatment (0.54 +/- 0.18 vs. 0.70 +/- 0.29; 0.26 +/- 0.13 vs. 0.41 +/- 0.25). HBV DNA became undetectable in 12 patients and decrease exceeded 100 times in 4 patients after treatment. HBeAg became undetectable in 10 of 14 patients who were HBeAg positive pretreatment patients, among them 2 patients had HBeAb sero conversion. The liver function was improved after treatment. All patients tolerated the treatment.

CONCLUSIONCIK treatment can increase immune effector cells and has some antiviral effect and is safe.

Adoptive Transfer ; adverse effects ; methods ; Adult ; Aged ; Cells, Cultured ; Cytokine-Induced Killer Cells ; cytology ; immunology ; transplantation ; Fatigue ; etiology ; Female ; Headache ; etiology ; Hepatitis B ; complications ; virology ; Humans ; Liver Cirrhosis ; etiology ; immunology ; therapy ; Male ; Middle Aged ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo evaluate the advantages of combination therapy with interferon-alpha plus nucleoside analogue-lamivudine or HBV vaccine in children with HBeAg positive chronic hepatitis B.

METHODSA total of 120 patients with HBeAg positive chronic hepatitis B were divided into three groups, 40 patients per group. Each group was treated with one of the following therapies respectively: Group A IFN-alpha 1b 10 MU/m2 three times per week (Tiw); Group B IFN-alpha 1b 10MU/m2 three times per week (Tiw) plus lamivudine 3 mg/kg for 6 months. Group C IFN-alpha 1b 10 MU/m2 three times per week (Tiw) plus HBV vaccine 30 microg one a month.

RESULTSThere was no significant difference in normalizing rate of ALT among the three groups at end of treatment. There was more significant difference in negative rate (seroconversion) of serum HBV DNA and HBeAg in group B than group A and group C (P less than 0.05).

CONCLUSIONThe combination therapy of IFN-alpha 1b plus lamivudine seemed to be more effective than the therapy with IFN-alpha alone and the combination of IFN-alpha and HBV vaccine.

Anti-HIV Agents ; therapeutic use ; Child ; Child, Preschool ; DNA, Viral ; blood ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Hepatitis B Vaccines ; therapeutic use ; Hepatitis B e Antigens ; blood ; genetics ; immunology ; Hepatitis B virus ; drug effects ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Lamivudine ; therapeutic use ; Male ; Treatment Outcome

OBJECTIVETo evaluate the quality of the ELISA diagnostic kits for detecting hepatitis E virus (HEV) specific IgG antibody.

METHODSFive diagnostic kits (WT, GD, HM, GeneLabs and KH) for anti-HEV IgG were assayed by detecting HEV IgG in the HEV diagnostic reference sera from 24 positive cases and 30 negative cases. 42 cases clinical suspect patients with hepatitis E from Ditan hospital in Beijing in 1994 to 1995 and 230 normal persons' sera from Chengdu city, Sichuan province in September in 2005.

RESULTSThe conformity rates of positive and negative sera was 95.83% (23) and 96.67% (29) for WT kit, 87.50% (21) and 100% (30) for GD kit, 87.50% (21) and 96.67% (29) for HM kit, 66.67% (16) and 100% (30) for GeneLabs kit, 45.83% (11) and 100% (30) for KH kit for detecting anti-HEV positive and negative reference sera, respectively. The five kits were used to detect anti-HEV IgG among 42 clinical suspect patients with hepatitis E and 230 normal persons' sera, The positive rate was 97.62% (41) and 31.74% (73) by WT, 100% (42) and 17.39% (40) by GD kit, 97.62% (41) and 23.91% (55) by HM kit, 90.48% (38) and 7.83% (18) by GeneLabs kit, 90.48% (38) and 3.48% (8) by KH, respectively.

CONCLUSIONThe positive rates of all the 5 kits were over 90% and had a very high conformity in detecting anti-HEV IgG in clinical patient with hepatitis E, positive rates of 3.48% to 31.74% were found in detecting the normal persons' sera. The research showed insufficiency of the ELISA kits for epidemiological survey of HEV infection in population.

Adolescent ; Adult ; China ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Hepatitis Antibodies ; blood ; Hepatitis E ; blood ; diagnosis ; virology ; Humans ; Immunoglobulin G ; blood ; Reagent Kits, Diagnostic ; standards ; Reproducibility of Results ; Sensitivity and Specificity ; Young Adult