Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38％ of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95％,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.

Objective To understand the potential risk factors influencing the effect of standard anti-tuberculosis (TB) treatment for TB patients co-infected with human immunodeficiency virus (HIV) and provide evidence for the improvement of anti TB therapy.Methods A retrospective study was conducted among 445 TB/HIV patients diagnosed and registered in 7 counties in Yunnan province from January 2010 to June 2012.A structured questionnaire was used to collect the patients' demographic characteristics,diagnosis and treatment information after informed consent.Chi-square test was conducted to compare successful rate of anti TB treatment among the patients with different demographic characteristics.Multivariate logistic regression analysis was conducted to identify risk factors influencing the effect of anti TB treatment.Adjusted OR＞1 means the risk factor of treatment failure.P value less than 0.05 was set as significant level.Results After standard anti TB treatment,397 patients were cured.The five risk factors influencing treatment effect were the existing of 4 suspected TB symptoms when seeking medical care for the first time(adjusted OR=2.208),TB/HIV patients detected in HIV/AIDS screening (adjusted OR=5.856),severe case (adjusted OR=4.607),non-full-course supervision during treatment (in intensive phase adjusted OR=4.129,full-course management adjusted OR=8.090) and interruption of therapy (adjusted OR=21.517).Conclusion Early detection of TB/HIV patients and conducting full course supervision during treatment can improve the effect of anti TB treatment.It is necessary to strengthen the early detection of TB/HIV patients and standarded treatment in Yunnan province.

OBJECTIVETo observe the inhibition of NK4 protein in the proliferation of human Raji lymphoma xenografts in nude mice, and to explore its molecular mechanism.

METHODSModels of human Raji lymphoma xenograft transfected with HGF gene were established by subcutaneous inoculation in nude mice. After establishment of the models, the mice received continuous NK4 protein via tail vein for 4 weeks, and the weight and tumor growth were monitored every week. After 8 weeks, the expression of HGF mRNA and c-Met mRNA of tumor tissues was measured by real-time fluorescent quantitation PCR. The apoptotic index (AI) and microvessel density (MVD) were evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry, respectively.

RESULTSThe models of human Raji lymphoma xenograft were successfully established. Although the animal weights of all groups declined, especially in the groups with NK4 protein injection, there was no statistical significance (P > 0.05). The tumor volume in HGF gene transfected group was larger than those of the control groups (P < 0.01), and there was no statistical significance among the control groups (P > 0.05). However, the tumor volume of the NK4 protein injection group decreased significantly (P < 0.01). Expression of HGF mRNA and c-Met mRNA in HGF gene transfected group increased significantly after injection of NK4 protein (P < 0.01). AI in HGF gene transfected group (33.5% ± 12.3%) was significantly lower than that of control groups (89.1% ± 22.3% vs. 81.9% ± 27.0%, P < 0.05), but became significantly higher (119.1% ± 18.9%) after NK4 protein injection (P < 0.01). MVD in HGF gene transfected group (28.5 ± 2.0) was higher than that of control groups (12.2 ± 1.4, 13.8 ± 1.3, P < 0.01), although declined (15.5 ± 2.5) after NK4 protein injection (P < 0.01).

CONCLUSIONSNK4 protein suppresses significantly the growth of human Raji lymphoma xenografts transfected with HGF gene. The pathogenesis may be involved in promoting tumor cell apoptosis and restraining tumor angiogenesis through competitive interrupting HGF/Met signal pathway.

Animals ; Apoptosis ; Hepatocyte Growth Factor ; genetics ; metabolism ; Heterografts ; Humans ; Lymphoma ; genetics ; metabolism ; therapy ; Mice ; Mice, Nude ; Microvessels ; pathology ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; T-Box Domain Proteins ; administration & dosage ; Transfection ; Transplantation, Heterologous