Objective To estimate the uncertainty of measurement in detecting of hepatitis B virus DNA(HBV DNA)by method of fluorescence quantitative polymerase chain reaction(FQ-PCR),and discuss the application value.Methods The process of the detection of HBV DNA by FQ-PCR was analysed to confirm and simplify the sources of uncertainties of measurement,which were obtained by disposing the data of methodology validation,internal quality control(type A evaluation of uncertainty)and external quality assessment(type B evaluation of uncertainty);combined uncertainty and expanded uncertainty were obtained by statistical methods.Results The main sources of uncertainties of measurement were:precision within laboratory,precision between laboratory,method bias.The expanded uncertainty of HBV DNA by FQ-PCR was U=0.62(k=1.96,n=2).The uncertainty caused by method bias was found mostly.Conclusion Expanded uncertainty can be compared in different results of HBV DNA by FQ-PCR,and it provides guide significance for observing the cure effect of anti-HBV and choosing the concentration of quality control.

267 newly-diagnosed patients with type 2 diabetes were divided into normoalbuminuria group [group N-UAlb, urinary albumin to creatinine ratio (UACR)<30 mg/g, n=103], microalbuminuria group(group M-UAlb, UACR 30~300 mg/g, n=88 ) , and macroalbuminuria group ( group L-UAlb, UACR>300 mg/g, n=76). The control group(group NC) consisted of 114 healthy individuals. Serum betatrophin, adiponectin(APN), and interleukin-1β( IL-1β) levels were determined with ELISA methods and the parameters of body mass index (BMI), estimated glomerular filtration rate(eGFR), HbA1C, fasting plasma glucose(FPG), OGTT 2h plasma glucose(2hPG), fasting insulin(FINS), OGTT 2h postprandial insulin(2hPINS), fasting C-peptide(FCP), homeostasis model assessment insulin resistant index(HOMA-IR), and blood lipid were collected. Compared with group NC, the serum betatrophin levels in patients with type 2 diabetes were obviously increased. In patients with type 2 diabetes, betatrophin levels increased along with the increase of UACR and there were significant differences in betatrophin among the three groups(P<0. 01). Betatrophin positively correlated with UACR, HbA1C, FPG, 2hPG, FINS, 2hPINS, HOMA-IR, TC, LDL-C, and TG( r were 0. 785, 0. 225, 0. 136, 0. 241, 0. 386, 0. 223, 0. 411, 0.216,0.193,and0.298,allP<0.05),and betatrophin were also positively correlated with APN and IL-1β(rwere 0. 643 and 0. 710, both P<0. 01). Stepwise multiple regression analysis showed that UACR, HbA1C, FINS, and TG were independent relevant factors affecting betatrophin levels.

Objective To evaluate the value of differential diagnosis on congenital biliary atresia(BA) and infantile hepatitis syndrome(IHS) by technetium-99m-diethyl-iminodiacetic acid(99Tcm-EHIDA)hepatobiliary scintigraphy with phenobarbitol sodium.Methods Fifty-eight infants with persistent jaundice were taken phenobarbitol sodium[5 mg/(kg?d)] ,bid ?7 d).Those who had not bowel and gallbladders radioactivity within 24 hours were diagnosed as the diagnostic criterion of BA.Those with bowel and gallbladders radioactivity within 24 hours were diagnosed as the diagnostic criterion of IHS,who then received 99Tcm-EHIDA hepatobiliary scintigraphy with single photon emission computed tomography(SPECT) instrument.The results of all children were analyzed and compared with pathology and clinical follow up results.Results 99Tcm-EHIDA hepatobiliary scintigraphy correctly diagnosed 24 infants with last diagnosis BA and 29 infants with last diagnosis IHS,5 neonates false positive in all 34 IHS patients.The sensitivity in the diagnosis of BA was 100%,the specificity and accuracy were 85.3% and 91.4%,restectively.The sensitivity was 85.3% in the diagnosis of IHS;the specificity and accuracy were 100% and 91.4%,respectively.Conclusions 99Tcm-EHIDA hepatobiliary scintigraphy with phenobarbitol sodium can accurately differentiate BA and HIS at early stage.

OBJECTIVETo study the association of the CD4+ cell counts and HBeAg with liver damage and cirrhosis in patients with chronic hepatitis B (CHB).

METHODTo measure the lymphocytes both CD4 and HLA-DR positive of 58 patients with CHB and 18 patients with HBsAg negative but HBcAb positive. The Platelets (PLT) were counted and the ALT and AST were measured, meanwhile, AST/PLT values were calculated.

RESULTSThe CD4+ cell counts of the three types patients were lower than those of healthy controls significantly,and those of HBcAb positive patients without CHB were higher than those of HBeAg negative patients with CHB which were higher than those of HBeAg positive patients significantly (P < 0.05). And the ALT, AST and AST/PLT levels of both HBeAg negative and positive patients were much higher than the three indicators of both HBcAb positive patients without CHB and healthy controls significantly (P < 0.02), meanwhile, the three indicators of HBeAg negative patients were much lower than those of HBeAg positive ones. In addition, the CD4+ cell counts of the two types patients were negatively correlative with the three indicators(P < 0.05).

CONCLUSIONDecreased CD4+ cell count can be used as the indicator to predicate the progress of liver damage in patients with CHB, and it is also useful for HBeAg to evaluate the development degree of liver damage and fibrosis in CHB patients.

Adult ; Aged ; Aged, 80 and over ; Alanine Transaminase ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; Female ; Flow Cytometry ; Hepatitis B e Antigens ; metabolism ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; etiology ; immunology ; Liver Diseases ; etiology ; immunology ; Male ; Middle Aged

Objective To investigate the HBV reactivation status and clinic outcomes in the renal allograft recipients with inactive HBsAg carriers,and explore the preventive measures.Methods A retrospective analysis of clinical manifestation was processed in 88 cases of inactive HBsAg carriers before and after renal transplantation.Preoperative liver function in all cases was normal and serum HBsAg positive,HBV DNA＜106 copies/L.Tacrolimus (or cyclosporine A) + mycophenolate mofetil (MMF) + prednisone were given in prevention of rejection after transplantation.In 88 cases,56 cases were given nucleoside analogues (acid) for prophylactic antiviral therapy,in which 31 cases were given lamivudine (LAM) (LAM group),25 cases were given entecavir (ETV) (ETV group) ; The rest 32 cases were not given prophylactic antiviral therapy,only receiving routine liver-protecting therapy (inosine,glucurolactone) (control group).Incidence of HBV re-activation,liver function,response to treatment and the pathological changes of hepatic tissue were observed.Results During the follow-up period,the incidence of HBV reactivation in LAM group and ETV group was 45.2％ and 28.0％ respectively,significantly lower than in control group (84.4％,P＜ 0.05).In prophylactic treatment groups,HBV reactivation occurred later,liver function damage was milder,and HBV DNA load peak was lower (P＜0.05).In LAM group,HBV reactivation occurred in 14 cases,including 10 cases occurred during administration of LAM,and ETV treatment was given for about 2 months,serum HBV DNA levels in 7 cases were under detection line;in the rest 4 cases,HBV reactivation occurred in patients with treatment less than 1 year and noncompliance,who withdrew medicine blindly.After the original scheme of antiviral therapy was done,serum HBV DNA levels in 3 cases were under detection line,and the effect was not obvious in one case.In control group,HBV reactivation occurred in 27 cases.Fourteen cases therefore accepted nucleoside (acid) analogs antiviral therapy,and HBV DNA levels in 10 cases were under detection line.Histological examination revealed the liver with fibrotic cholestatic hepatitis changes in 9 patients,including 8 cases in control group,and 1 case in LAM group due to blind withdrawal of medicine.Conclus(i)on LAM and ETV prophylactic use may decrease the HBV reactivation rate in inactive HBsAg carriers after renal transplantation,reduce the severity of liver damage and the occurrence of fibrotic cholestatic hepatitis.

Objective For Genistein has been reported to inhibit many tumors ,we investigate the role of autophagy in the proliferation inhibition to Hela cells by Genistein and the machanism of autophagy plays in this process.Methods Human cervical cancer Hela cells were divided into control group,Genistein group and 3-MA+Genistein group,the control group were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum(FBS),Genistein group were cultured in various concentrations Genistein(25,50,100μmol/L),3-MA+Genistein group were treated with 5mmol/L 3-MA for 1h before cultured in 100μmol/L Genistein.The proliferation inhibitory rate of Hela cells was detected by MTT method.The ultrastructure changes of Hela cells was observed under transmission electronic microscope(TEM).The levels of autophagy-associated protein P62 and Beclin-1 were detected by Western blotting analysis.The expressions of autophagy-associated proteins LC3A/B in Hela cells were determined by fluorescent staining to analyse the autophagy induced by Genistein in Hela cells.Results Compared with control group ,the proliferation inhibitory rate of Hela cells was 20.9%±1.3%,33.5%±1.6% and 46.5%±3.2% when cultured in 25,50,100μmol/L Genistein(P<0.01).After treated with various concentrations Genistein for 48h, we observed a dose-dependent increase in the expression of Beclin-1 and decrease of P62.Confocal laser scanning microscopy confirmed the fluorescent density of LC3A/B expression in Hela cells treated with 100μmol/L Genistein increased significantly as compared with control group.TEM showed there are many vacuoles and double-membrane autophagosomes which involved cytoplasmic components in Hela cells treated with 100μmol/L Genistein.The proliferation inhibitory rate of Hela cells of Genistein group is decreased as compared with those in 3-MA+Genistein group[(46.5±3.2)% vs (58.2±2.2)%,P<0.01].Conclusion Genistein could inhibit Hela cells proliferation and induce autophagy.

Aim To investigated the possible effect of COX-2 on the BMP9-induced activation of PI3K/Akt signal in progenitor cells.Methods The activity of alkaline phosphatase(ALP) was measured using histochemical staining or chemiluminescence.The mRNA level of ALP was determined using real-time PCR assay.The protein levels of osteopontin(OPN), osteocalcin(OCN), COX-2, Akt1/2 and phosphorylated Akt1/2 were detected by Western blot.The mRNA level of COX-2 was assayed with RT-PCR, and the mineralization was measured with Alizarin Red staining.Results The ALP activity was apparently increased by BMP9 in C2C12 cells, as well as the protein level of OPN and OCN.The mineralization was also markedly induced by BMP9 in C2C12 cells.BMP9 increased the level of phosphorylated Akt1/2 greatly, although no substantial effect was observed on total protein level of Akt1/2.The BMP9-induced ALP activity was dramatically decreased by the inhibitor of PI3K.The mRNA and protein level of COX-2 were both increased by BMP9 in C2C12cells, and the BMP9-induced ALP activity and mineralization were greatly attenuated by the inhibitor of COX-2.The BMP9-induced phosphorylation level of Akt1/2 was increased by the exogenous expression of COX-2, but decreased by the inhibitor of COX-2.Conclusion Activation of PI3K/Akt signaling may be a critical event in BMP9-induced osteogenic differentiation, and this process may be mediated by the BMP9-upregulated COX-2 in stem cells at least.

The positive ratio is 74 percent using the plate anaerobe culturing device made by author, higher than using common anaerobic box and anaerobic jar.

Objective To explore the effects of intestinal trefoil factor ( ITF) on gastric mucosal epithelial cell proliferation and its possible molecular mechanism . Methods The cultured GES-1 cells were treated with ITF in the concentration of 100 ng/mL and 500 ng/mL in vitro, and then were observed using microscope for the morphological analysis .The Cell Counting Kit-8 ( CCK-8) was used to detect the proliferation activity of GES-1.The cultured GES-1 cells were treated with 100 ng/mL ITF and the specific inhibitor of PI3K/Akt signaling pathway LY294002 (15μmol/L) in vitro, and then were observed using microscope for the morphological analysis . The proliferation activity of treated GES-1cells was detected using CCK-8 and the expressions of p-Akt and Akt of PI3K/Akt signaling pathway were determined by Western blot . Results Compared with the control group , the proliferation activity of GES-1 cells in-creased after being treated with ITF and the higher concentration of ITF induced the higher proliferation activity .LY294002 inhibited the increased proliferation activity of GES-1induced by ITF.The data of Western blot indicated that ITF induced the expression of p -Akt and activated the P3IK/Akt signaling pathway to modulate the proliferation activity of GES -1 cells.However, LY294002 inhibited the PI3K/Akt signaling pathway and then decreased the proliferation activity of GES -1 cells. Conclusion ITF could promote the proliferation ac-tivity of GES-1 cells by activating PI3K/Akt signaling pathway .

Objective To assess the impact of the virus on the complementary determining region 3 (CDR3) length diversity of T cell receptor(TCR) Vβ repertoires of CD4+ T lymphocytes and to explore its association with viral load in individuals with HIV-1 infection. Methods The TCR repertoire was examined using spectratyping of CDR3 length diversity within CD4+ T cells in HIV infected and healthy adults. Separation of CD4+ T cells from peripheral blood mononuclear cells ( PBMCs) was carried out by using immunomagnetic beads coated with anti-CD4 antibody. Total RNAs from the purified CD4 + T lymphocytes were isolated and used to perform nested-PCR amplifications in CDR3 of 22 TCR gene families. CDR3 diversity and its association with viral load in individuals with HIV-1 infection were analyzed. Results An average diversity for all CDR3 profiles in CD4+ T cells from 25 HIV-infected individuals was significantly different as compared to 10 age-matched healthy donors (P＜0.05) with the HIV-infected individuals losing diversity in the CDR3 profiles. There was positive correlation between changes in TCR CDR3 diversity and viral load (r = 0. 494, P ＜ 0. 05). The changes in CDR3 length diversity of Vβ families in HIV-infected individuals, particular in Vβ8, Vβ22, Vβ23 were statistically different from the healthy controls. Conclusion HIV-1 infection might induce the loss of TCR Vp repertoire diversity and disrupt the CDR3 Gaussian distributions within CD4 + T cells. There should be positive correlation between changes in TCR CDR3 diversity and the viral load in HIV-1 infected patients.