ABSTRACT
Objective To indentify the relationship between hepatitis B virus covalently closed circular DNA (HBV cccDNA) and postoperative recurrence of HBV in HBcAb positive liver donors by detecting HBV cccDNA in liver from HBcAb positive liver donors.Method Eighty-five of 1200 patients underwent liver transplantation for hepatitis B-related end-stage liver disease in our hospital from January 2007 to January 2010 were retrospectively analyzed.According to the situation of HBV infection in donor liver,the recipients were divided into 3 groups:(1) the experimental group (livers positive for HBcAb,and negative for HBsAg,n =40),(2) control group (livers positive for HBsAg,n =15),and (3) normal group (donor livers without HBV infection,n =30).HBV cccDNA of donors was detected by fluorescence quantitative PCR.Serum HBV and HBV DNA were regularly tested,and biopsy was done in those positive for HBsAg or HBV DNA to confirm HBV recurrence.The relationship between HBV cccDNA and postoperative recurrence of HBV in HBcAb positive liver donors was analyzed.Result The positive rate of cccDNA was 30％ in experimental group (12、40),73.3％ in control group (11/50),and 0 (normal group).The rate of HBV recurrence in experimental,control and normal groups was 10％ (4/40),80％ (12/15) and 3.3％ (1/30) respectively.The rate of HBV recurrence in the experimental group of cccDNA (+) and cccDNA (-) was 7.5％ (3/40),and 2.5％ (1/40).Conclusion The cccDNA in HBcAb positive donors may be one of the risk factors in HBV recurrence of hepatitis B-related liver transplantation patients.The screening of HBV cccDNA in the donor livers positive for HBcAb before liver transplantation is recommended to reduce the positive HBV recurrence and expand the pool of liver donors for patients with HBV-related liver disease.
ABSTRACT
Objective To establish a high-performance induction culture system for Raw264.7 cells to differentiate into osteoclasts(OC)invitro by improving the cell culture program.Methods The Raw264.7 cells were cultured withα-MEM medium containing 50 μg · L-1 M-CSF, 100 μg · L-1 RANKL, and 1 × 10-8 mol · L-1 1α,25-(OH)2 D3 in 5% CO2 for 12 d at 37℃. The cells were digested transiently every time before the medium was changed after every three days. The morphologic changes of the Raw264.7 cells were observed by inverted microscope.The maturation and phagotrophic function of OC were identified by HE,TRAP,FITC-phalloidin staining and immunofluorescence.Results The cells remained to grow in single layers all the time in most areas of the well during the whole induction by the improved culture program. The observation results of inverted microscope and HE staining showed that the growth area of the polykaryotic OC reached to 70% of the well on day 1 2. FITC-phalloidin staining showed that in the maturation of the OC, the cluster-shaped podosomes in the pseudopodia gradually transformed into rings,which finally fused to form a large belt surrounding the periphery of the cytoplasm. The calcitionin receptor (CTR) expressed by OC was markedly enhanced compared with the precursor cells by immunofluroescence staining,and a large number of red granules appeared in the cytoplasm of OC with TRAP staining on day 1 2. These results comfirmed that the obtained OC were maturated and owned phagotrophic function. Conclusion A high-performance induction culture system for Raw264. 7 cells to differentiate into OC in vitro induced by combination of 50μg · L-1 M-CSF, 100 μg · L-1 RANKL,and 1 × 10-8 mol·L-1 1α,25-(OH)2 D3 is established by improving the cell culture program.
ABSTRACT
Objective To analyze the clinical diagnosis and treatment strategies of liver-localized posttransplantation lymphoproliferative disease (LL-PTLD).Methods Six cases of LL-PTLD from more than 3000 cases of liver transplant recipients from July 2003 to July 2011 were retrospectively analyzed.Other six cases of LL-PTLD were retrieved through Pubmed and Wanfang.The diagnosis and treatment of 12 cases of LL-PTLD were summarized and analyzed.Results All patients with LL-PTLD were diagnosed pathologically.The incidence of LL-PTLD was 0.2％ (6/3000).Among 12 patients,immunosuppressant and anti-EB virus treatment was reduced or withdrawn in the vast majority of patients,and treatment response was satisfactory.Systemic chemotherapy was given in 6 cases,and three of them died.Local radiation therapy was given in 4 cases,the tumor was significantly controled,and patients survived.Secondary liver transplantation was performed on 3 cases: 1 case died of recurrent lymphoma,and one case received partial hepatectomy and no lymphoma recurred.Conclusion For cases with obstructive symptoms of fever and chills associated with jaundice without reasonable explanation,LL-PTLD is suspected and diagnosed by liver biopsy.Basic treatments such as adjustment of immunosuppressive agents and anti-viral therapy are recommended as early as possible.Local radiation therapy is a treatment method of LL-PTLD,which can obtain a satisfactory therapeutic effect.