目的:本项目采用焦磷酸测序法对慢性丙型肝炎（CHC）患者的IL-28B基因SNP进行分型,明确CHC患者中IL-28B等位基因频率与其中医证候、肝纤维化的分布特征,借以比较肝纤维化在不同IL-28B等位基因及中医证候患者中的发生率,并指导今后针对CHC患者尤其是可能进展为肝硬化的高危患者的治疗。方法:采用焦磷酸测序法对在我院门诊及住院的105例CHC初治患者的IL-28B基因rs12979860 SNP位点的基因型进行检测,完善其血生化、病理学等检查,再对其进行中医辨证分型,比较不同等位基因中的中医证候的分布及不同证候中肝纤维化患者所占的比例。结果:采用焦磷酸测序法测得我院CHC患者的IL-28B基因分型为CC型（90例）及CT型（15例）。在等位基因为CC型的CHC患者中,中医证候为肝肾阴虚的患者比例最多[41.3%（37/90）],而CC型患者中肝纤维化的比例为53.3%（48/90）,CT型为20%（3/15）,差异有显著性意义（＜0.05）,其中CC型患者中医证候为肝肾阴虚的发生肝纤维化的相对危险度RR值为4.1,95%CI为1.6~10.2。结论:IL-28B等位基因为CC型的CHC患者主要中医证候为肝肾阴虚,CC型比CT型更容易发生肝纤维化,CC型中肝肾阴虚患者发生肝纤维化的概率最高,我们应加强对IL-28B等位基因为CC型患者的治疗与监测。

Objective To study the effect of total saponins of panax notoginseng(PNS) on the expression of bone morphogenetic protein-2(BMP-2) and alkaline phosphates(ALP) activity in rat bone marrow stromal cells(BMSCs),and to elucidate the mechanism of PNS action on the proliferation of BMSCs during differentiation into osteoblasts.Methods After isolation and purification,rat BMSCs were induced in medium alone(control) or osteogenic medium containing different concentrations of PNS(50,100,150 and 200 mg·L-1).The proliferation of BMSCs was determined by MTT assay.The osteogenic differentiation was evaluated by ALP activity.BMP-2 expression was detected by Western blotting.Results There were no significant differences in the proliferation of BMSCs at 3 and 5 days after treatment among the different groups(P0.05).Compared with control group,the proliferation of BMSCs,BMP-2 expression and ALP activity were significantly increased by treatment with 50,100 or 150 mg·L-1PNS at 7 and 9 days after treatment(P0.05).However,no significant differences were found in cell proliferation,BMP-2 expression and ALP activity between 200 mg·L-1 PNS treatment group and control group(P0.05).Conclusion The PNS can promote the differentiation of BMSCs into osteoblasts through increasing BMP-2 expression and ALP activity.

Objective To construct the recombinant prokaryotic expression vector containing the Early secretary antigenic target(ESAT-6) gene of Mycobacterium tuberculosis,expression the fusion protein identified.Methods We cloned the ESAT-6 gene from mycoba-cterium tuberculosis by PCR,into the expression Vector pET-32a(+)with the BamH Ⅰ,Hind Ⅲ two enzyme sites,identified the recombinant vector by PCR,sequencing.expressed the protein in E coil.BL21,identified the fusion protein with Western-blotting.Results The sequencing result of ESAT-6 gene was the same as the Genbank.The relative molecular mass of target protein is about 2.5×10~4,has the activity of banding antibody.Conclusion The ESAT-6 fusion protein was successfully expressed and purified,identified.