Objective To evaluate the two methods for quantitative hepatitis B virus (HBV) DNA test. Methods The Hybrid Capture II system from Digene Co. and Real Time PCR fluorimetry quantitative HBV DNA test kit from PIJI Bio Technical Development Company Ltd. were used to detect sera HBV DNA in sera of patients with chronic hepatitis B. Results With the two quantitative methods, the positive rates of HBV DNA were 98 6% and 94.6% respectively. The concordant rate of these two methods was 93.2%. In assessing the sensitivity of the two kits with a serial of 10 fold diluted patients′s sera, PG kit could test 10 -7 of diluted serum, and HC II could test 10 -6 . The test value of HC II was more accurate in quantitation than PG kit especially in higher titer of HBV DNA. Whereas in the case of low titer of HBV DNA, the deviation of test value increased in both two methods. For monitoring the anti virus effect with quantitative HBV DNA in CHB patients, these two methods had same sensitivity, and the test value of HBV DNA had the same trend of change. Conclusion HC II system and PG test kit were sensitive and reliable, and they were worthy of monitoring the anti virus efficacy and of clinical practice.

OBJECTIVETo study whether the abilities of hepatitis C virus (HCV) E2 gene immunization to induce humoral and cellular immune responses to E2 protein were affected by hepatitis B virus (HBV) preS gene when they fused in DNA-immunized mice.

METHODSMice were immunized with E2, preS-E2 (preS gene was upstream of E2 gene), and E2-preS (preS gene was downstream of E2 gene) gene by their eukaryotic expression vectors, respectively. The anti-E2 or anti-preS antibodies were detected using the E2 and preS antigens. The cellular immune response to E2 protein in immunized mice was presented by its survival time after injecting SP2/O myeloma cells expressing HCV E2 protein into the abdominal cavity.

RESULTSChimeric E2 and preS gene immunization can induce mice to develop anti-preS and anti-E2 antibodies. The number of the mice developing anti-E2 antibody and the antibody titers in preS-E2 gene-injected group were higher than those in E2-preS gene-immunized group. However, the mice injected with E2 gene did not develop the detectable anti-E2 antibodies until 12 weeks after DNA immunization. After the mice was injected with target cells, the average survival time of the mice in the group immunized with E2 gene alone was longer than that of the group injected with E2 gene fused with HBV preS and was significantly longer than that of the control (P < 0.05).

CONCLUSIONHBV preS might be a humoral enhancer that can affect the abilities of HCV E2 protein to induce immune responses in DNA-immunized mice.

Animals ; Antibody Formation ; drug effects ; Hepacivirus ; chemistry ; Hepatitis B Surface Antigens ; genetics ; pharmacology ; Hepatitis B virus ; chemistry ; genetics ; Immunity, Cellular ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Protein Precursors ; genetics ; pharmacology ; Recombinant Fusion Proteins ; immunology ; Viral Envelope Proteins ; genetics ; pharmacology

Objectives To sequence E2/NS1 gene from genotype Ⅲ Chinese isolates of hepatitis C virus (HCV) and analyze homology corresponding to the region of the reported isolates.Methods E2/NS1gene derived from genotype Ⅲ Chinese isolates of HCV was amplified by reverse transcripase-polymerase chain reaction (RT-PCR) and cloned into vector pcDNA3. Dideoxy chain termination methods were used to sequence E2/NS1 gene. Results E2/NS1 gene derived from genotype Ⅲ Chinese isolates of HCV was cloned for the first time and named HC-W14. Identity of HC-W14 in nucleotide and putative amino acid to those of genotype Ⅲ Japanese isolates of HCV were 88.37% and 89.29% respectively. Homology to that of non-type Ⅲ isolates was relatively low.Conclusions High variation existed in E2/NS1 region between genotype Ⅲ and Ⅱ Chinese isolates of hepatitis C virus. The variability of E2/NS1 gene should be taken into account in the development of vaccine against HCV in China.

Objective　To find neutralizing antibody candidates against hepatitis C virus (HCV) infection. Methods　We constructed two eukaryotic expression vectors which contained the E1 and E2 gene of HCV, and detected their expression in mammalian cells with transient expression. BALB/c mice were given subculaneous injections of constructed vectors combined with the IL-2 gene intraepidermally and evaluated for induced humoral immune responses by enzyme-linked immunosorbent assay (ELISA). We used an antibody-virus interaction assay to analyze the interaction of the antisera and HCV viral particles in vitro. Results　Anti E1 and anti-E2 antisera were obtained from immunized mice. The serum of mice immunized with the E2 gene immunoprecipitated the HCV isolate in source serum and reacted with the isolates unrelated to the original one. Conclusions　Anti-E2 antibody in induced mice can cross-reactively capture HCV particles, highlighting the possibility of generating broadly reactive anti-E2 antibodies.

OBJECTIVETo summarize the clinical characteristics, early diagnosis, comprehensive treatment and prognosis of 6 cases of children with post-transplantation lymphoproliferative disorder (PTLD) after liver transplantation.

METHODData of 6 cases with PTLD seen between January 2011 and December 2013 were retrospectively analyzed. The anti-rejection drug dose adjustments, the effect of rituximab, antiviral therapy and comprehensive treatment program after surgery were explored.

RESULT(1) The diagnosis of PTLD was confirmed by histologic findings. Six cases of PTLD including 3 males and 3 females were diagnosed as congenital biliary atresia and underwent split liver transplantation. The occurrence rate of PTLD was 2.9%. (2) The median time to the development of PTLD was less than 6 months. The initial symptom of PTLD in all patients was fever and clinical manifestations of PTLD were non-specific, depending on the involving organs. Five cases of PTLD developed gastrointestinal symptoms, including diarrhea, abdominal pain, and abdominal distension. One case developed respiratory symptoms, including cough and tachypnea. Three cases had lymph node involvement. In 2 cases pathophysiology involved polymorphic lymphocyte proliferation and in 4 cases B lymphocyte proliferation. (3) Two cases died, in whom EBV DNA was not detected and were diagnosed as PTLD by surgical pathology before death. Four survived cases had high EBV-DNA load and then were diagnosed as PTLD by biopsy pathology. (4) Of the 6 cases of PTLD, 2 cases died and 4 cases survived. The overall mortality was 33%. The dead cases were only treated with laparotomy because of intestinal obstruction or perforation and the survived cases were treated with tacrolimus at reduced doses or discontinuation and rituximab. In 2 cases antiviral therapy (acyclovir) was continued, including 1 cases of intestinal obstruction treated with surgical repair. All the survived patients were followed up for 4 months to 1 year and no evidence has been found.

CONCLUSIONEBV infection is the high risk factor for PTLD after liver transplantation. Close clinical surveillance of EBV DNA for pediatric liver transplantation was important for the early diagnosis of PTLD. Reducing doses of immunosuppressive agents and rituximab is the initial therapy for PTLD. A reduction in the dose of tacrolimus is suggested. Operation therapy can also play a role in the management of local complications.

Antiviral Agents ; administration & dosage ; Biliary Atresia ; therapy ; DNA, Viral ; analysis ; Drug Therapy, Combination ; Early Diagnosis ; Epstein-Barr Virus Infections ; diagnosis ; therapy ; Female ; Humans ; Immunosuppressive Agents ; administration & dosage ; adverse effects ; Infant ; Liver Transplantation ; adverse effects ; Lymphoproliferative Disorders ; diagnosis ; etiology ; mortality ; therapy ; Male ; Pediatrics ; Postoperative Complications ; Retrospective Studies ; Survival Rate ; Tacrolimus ; administration & dosage