Objective:To determine the effects of Foxp3-overexpressing lung cancer cells on activated CD4+T lymphocyte.Methods: Stable Foxp3-overexpressing lung cancer cells NCIH-1299,NCIH-hFoxp3,was generated by transfection of NCIH-1299 cells with plasmid pcDNA3-hFoxp3 mediated by Lipofectamine 2000 and by selection with G418,and validated by quantitative PCR and Western blot.The expression levels of IL-8 and IL-10 secreted by NCIH-hFoxp3 and NCIH-control were measured by ELISA.IL-2 secrection by activated human CD4+T lymphocyte which was tested after stimulation with 20% conditioned medium of NCIH-hFoxp3 and NCIH-control cells.The proliferation of activated human CD4+ T lymphocytes was assessed by MTT after coculture with NCIH-hFoxp3 cells.The adhesive ability of activated human CD4+ T lymphocytes was probed with NCIH-hFoxp3 cells by immunocytochemistry.Results: Compared with NCIH-control cells,NCIH-hFoxp3 secreted high level of IL-10 and low level of IL-8.NCIH-hFoxp3 with Foxp3 overexpression significantly suppressed the proliferation,adhesive potential and IL-2 expression by activated CD4+ T cells.Conclusion: Suppression of immune activities of activated CD4+ T cells by Foxp3 overexpression in lung cancer cells may correlate with cytokine IL-8 and IL-10,which can contribute lung cancer progression.

Objective:To determine the effects of enforced expression of Foxp3 in lung cancer cell with regards to proliferation and tumorgeneity. Methods: A stable subline NCIH-hFoxp3 was established by liopfectamin-mediated pcDNA plasmid transfection carrying exogenous hFoxp3. The growth curve and secrection of IL-8 and IL-10 of NCIH-hFoxp3 were evaluated using MTT and ELISA, respectively. The in vivo tumorigeneity was assessed as well by inoculation of NCIH-hFoxp3 subcutaneously in nude mice. Results:Lung cancer cell NCIH-hFoxp3 with enforced expression of Foxp3 proliferated slowly but exihited increased in vivo tumorgeneity compared with corresponding control subline. In addition,increased expression of hFoxp3 in NCIH-hFoxp3 augmented secretion and at-tenuated secretion of IL-8 and IL-10,respectively. Conclusion:Increased expression of Foxp3 may promote progression of lung cancer cell by change of cellular microenvironment and evasion of immune surveillance.

Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.

Objective To explore the biological function of rhoptry protein 38(ROP38)of Toxoplasma gondii,and to iden?tify the reactogenicity of the recombinant protein(rROP38). Methods The ROP38 was amplified by RT?PCR from T. gondii RH strain,and was cloned into prokaryotic expression vector pET?28a(+). The recombinant plasmid was transformed into E. co?li BL21(DE3)competent cells. Then the rROP38 was analyzed by SDS?PAGE and identified by Western blot. Results SDS?PAGE showed that rROP38 was efficient expression with a molecular weight of about 43 kD. Western blot showed that rROP38 reacted with antibody of His tag or human positive antibody,which indicated that ROP38 had good reactogenicity and could be a serological diagnostic antigen. Conclusion The study successfully obtains the rROP38 of T. gondii with good reactogenicity.

OBJECTIVETo investigate the polymorphisms of 17 Y-STR loci in Han population in Guangdong Province and explore its application in forensic medicine.

METHODSeventeen Y-STR loci (DYS19, DYS385a/b, DYS635, DYS389I, DYS389II,DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, Y-GATA-H4) were analyzed using Y filer™ kit in 1000 unrelated individuals and 1000 DNA-confirmed father-son pairs. All mutations were confirmed by DNA sequence analysis.

RESULTSNo shared haplotypes were observed among the 1000 unrelated male individuals, and the gene diversity of 17 Y-STR loci ranged from 0.4285 (DYS391) to 0.9654 (DYS385a/b). The samples yielded an overall haplotype diversity of 1.000. A total of 17000 meiotic events were investigated in 1000 father-son pairs with DNA-confirmed biological paternity and 46 differences were observed between the fathers and sons. The average mutation rate was 0.0027 (95%CI, 0.0020-0.0036) per locus.

CONCLUSIONThe 17 Y-STR loci included in Y-filer have high genetic polymorphisms in South China Han population and have good prospect for application in forensic medicine.

China ; Chromosomes, Human, Y ; genetics ; Ethnic Groups ; genetics ; Gene Frequency ; Haplotypes ; Humans ; Male ; Polymorphism, Genetic