ABSTRACT
Objective To establish and assess a new screening method for anti-HIV-1 drugs.Methods JLTRG cells were co-cultured with different proportions of H9/HTLV-Ⅲ B cells for 24,48,72 and 96 h.Intensity and density of green fluorescent protein were observed under fluorescence microscope,and were tested using flow cytometry.The optimal proportion of cells in co-culture system and the culture time were determined.The effectiveness of Enfuvirtide (T20) and Efavirenz (EFV),and their half maximal inhibitory concentrations (IC50) were determined by using cell co-culture system and half life of drugs.HIV load was detected using RT-PCR for HIV-1 p24 antigen,and its correlations with drug concentration and mean fluorescent intensity were analyzed by Spearman rank correlation analysis.Results Experiments demonstrated that JLTRG cells co-cultured with H9/HTLV-ⅢB cells at the proportion of 10 ∶ 1 for 72 hours was the best.Along with the concentrations of T20 and EFV changed,JLTRG cells were infected with HIV-1 in different degrees,and the IC50s of T20 and EFV were 10 nmol/L and 5 nmol/L,respectively.The concentrations of T20 and EFV were negatively correlated with mean fluorescent intensity and viral load (r =-1,-0.986 and-1,-1,P ＜ 0.01); and mean fluorescent intensity was positively correlated with viral load (r =0.986 and 1,P ＜ 0.01).Conclusion The drug screening method established in this study is efficient and easy to operate,which provides a new option for anti-HIV-1 drug screening.
ABSTRACT
OBJECTIVE To investigate the infection and drug resistance of Staphylococcus spp from hospitalized cases in Suzhou area.METHODS The data from hospitalized cases of 32 hospitals in Suzhou(from 2004 to 2007) were analyzed retrospectively.RESULTS From 2004 to 2007,17 668 cases of nosocomial infection were collected from 32 hospitals in Suzhou area.The infection rate of Staphylococcus aureus was 5.78%,7.11%,8.39% and 7.50%,respectively;the number of meticillin-resistant S.aureus(MRSA) infection cases was 66(34.74%),107(33.86%),138(37.00%) and 219(53.16%) respectively and the total number was 530(41.05%).The nosocomial infection caused by S.epidermidis accounted for 5.99%,5.47%,5.35% and 5.25%,respectively from 2004 to 2007.The number of meticillin-resistant S.epidermidis(MRSE) infection cases were 118(59.90%),128(52.67%),119(50.00%) and 134(46.53%) and the total number was 499(51.66%).The main infection site of S.aureus and S.epidermidis was respiratory tract(74.28% and 71.81%,respectively).Antibiotic resistance strains of S.aureus and S.epidermidis was on rising,and most of them were multi-drug resistance strains.All of the strains were sensitive to vancomycin.CONCLUSIONS In Suzhou area,nosocomial infection and drug resistance of Staphylococcus is on the rise.Evevy hospital must take effective measures to control nosocomial infections of Staphylococcus and drug resistance.
ABSTRACT
This study sought to isolate and purify C-kit+ cells from rat 2-AAF/PH model and to investigate the proliferation and differentiation of C-kit+ cells in vitro. C-kit positive oval cells were enriched by using magnetic activated cell sorting (MACS). The sorted oval cells were cultured in a low density, and then colony formation was observed. The capacity of proliferation and differentiation of C-kit positive cells were examined in vitro by immunocytochemistry and RT-PCR. By using C-kit antibody in conjunction with MACS, we developed a rapid oval cell isolation protocol. The sorted cells formed colony when cultured in vitro. Cells in the colony expressed albumin or cytokeratin 19 (CK19) or coexpressed both, and BrdU incorporation test was positive. RT-PCR on colony showed expression of albumin and CK19 gene. The results demonstrate that by means of MACS we have established a method to isolate oval cells. The sorted hepatic oval cells can form colony in vitro which expresses different combinations of phenotypic markers and genes from both hepatocytes and cholangiocyte lineage.