ObjectiveTo optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.MethodsFour HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.ResultsBoth the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.ConclusionAllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.

Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P

Objective To evaluate hepatitis B virus large surfsce protein(LHBs) in monitoring of antiviral treatment.Methods LHBs.HBV serum markers and HBV DNA loads in 46 patients with adefovir dipivoxil(ADV) therapy were monitored for the whole course(60 weeks).Enzyme linked immunosorbent assay(ELISA),time differentiate immunofluoresence assay and real.time polymerase chain reaction(RT-PCR)were performed to detect LHBs,HBV serum markers and HBV DNA loads,respectively.And correlation analysis was also performed.Results Both LHBs and HBV DNA declined during the ADV treatment.and the correlation coefficient was 0.97.but LHBs declined after HBV DNA.There were 20 patients with HBV DNA<5×102/mL at 60th week.in which 8 were LHBs negative;in 14 recurrent patients,the HBV DNA turned to negative and became positive again,3 with negative LHBs;while in 12 patients resistant to the ADV therapy.2 were LHBs negative.Conclusion Dynamic monitoring of LHBs is useful in the evaluation of antiviral treatment.

To observe the effects of a Chinese compound herbal medicine for reinforcing qi, activating blood circulation, expelling phlegm and dredging collaterals in treatment of lower limb arteriosclerosis.

ObjectiveTo investigate the clinical characteristics,epidemiology of patients with severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) infection and genetic sequences of SFTSV.MethodsClinical data of five cases of severe fever with thrombocytopenia syndrome (SFTS)from Zhoushan Hospital during May 2011 to July 2011 were retrospectively analyzed.SFTSV gene was amplified by polymerase chain reaction (PCR).CD3+ CD4+ and CD3+ CD8+T lymphocytes were detected by flow cytometry (FCM).The sequences of isolated SFTSV strains were compared with those in GenBank. ResultsThe symptoms of continuous high fever,sore muscles,enlarged superficial lymph nodes,abdominal pain,diarrhea with gastrointestinal hemorrhage were observed.The white blood cells,platelets and CD3+ CD4+ T lymphocytes were progressive decreased in acute phase with the minimum of (0.97-2.00) × 109/L,(12-42) × 109/L and 7.52％-20.39％,respectively.The SFTSV was isolated from the sera of two patients.The sequences were compared with SFTSV sequences in GenBank.The homology of RNA-dependent RNA polymerase gene was 96％ compared with BX-2010,L-WWG,LN3,JS4,SD4,HN6 and AH12; the glycoprotein gene was 94％ ; N protein gene was 95％ compared with JS4,SD4 and LN4.The homology of the above three genes between two isolates was 99％.ConclusionsOur results suggest that SFTSV is sporadic in Zhejiang Province which is probably from native epidemic focus.SFTS is progressive and severe with acute onset.Multiple organ dysfunction is common in severe eases.