1.Cloning And Expression Of Mtb8.4-Hspx And Hspx-Mtb8.4 From Mycobacterial Tuberculosis 2.FTY720 Immunosuppression For Kidney Transplant Recipients

Objective To construct recombinant vector pET30a-Mtb8.4-Hspx(pET30a-MH) and pET30a-Hspx-Mtb8.4 (pET30a-HM) without affinity tag and express them in E. coli BL21 (DE3) strain.Method The two genes encoding protein Mtb8.4 (containing Ndeâ… and Sacâ… ) and Hspx (containing Ndeâ… and Sacâ… ) were amplified from M. tuberculosis H37Rv chromosomal DNA using PCR. The two amplification products were purified and then inserted into plasmid vector pET30a digested with Ndeâ… and Sacâ… to construct recombinant plasmid pET30a-Mtb8.4,pET30a-Hspx. The two recombinant plasmids were transferred into E. coli DH5a strain. Extracting Plasmid DNA (pET30a-Mtb8.4, pET30a-Hspx) and digesting them with Sacâ… and Hindâ…¢, and then purifying them. Amplifying Hspx (digested by Sacâ… and Hindâ…¢) and Mtb8.4 (digested by Sacâ… and Hindâ…¢) and constructing recombinant plasmid pET30a-Mtb8.4-Hspx (pET30a-MH) and pET30a-Hspx-Mtb8.4 (pET30a-HM). The products were identified by DNA sequence and SDS-PAGE. Plasmid containing the right inserted fragment was retransformed into E. coli BL21 (DE3) strain.Results The recombinant products MH and HM without any affinity tag apparent Molecular Weight are 28 000, and they had been identified by SDS-PAGE.Conclusion The successful expression of MH and HM protein without affinity tag has made a fundament for the research of immunogenicity of Mycobacterium Tuberculosis vaccine. Objectives To compare the positive and negative effects of FTY720 and MMF as maintenance immunosuppression therapy for kidney transplant recipients.Methods We comprehensively searched Medline (1966-October 2010), Embase (1980-October 2010), Cochrane Library (2009,4th), VIP(1989-2010), CNKI (1994-2010).Results Four studies (1883 patients) were included. At six and twelve months, the occurrence of first treated biopsy-proven acute rejection (BPAR) was significantly increased in [FTY720(5.0mg)+RDC] treated recipients (RR=1.45,95% CI [1.17 to 1.79]) but did not vary with [FTY720 (2.5mg)+FDC] groups (P=0.32, P=0.60) respectively compared with (MMF+FDC). At six month, there is no significant difference between FTY720 (2.5mg)+FDC treated patients and MMF+RDC group in the death rates of patients (RR=0.29,95% confidence interval 0.04 to 2.17), the rate of'graft loss'(RR=1.31,95% confidence interval 0.56 to 3.08) and the rate of first BPAR treated with antibody therapy(RR=0.65,95% confidence interval 0.27 to 1.57). In terms of the adverse effect, FTY720 caused more viral infection (RR=0.59,95% confidence interval 0.38 to 0.94), more bradycardia (RR=3.58,95% confidence interval 2.08 to 6.24), but there is no significant difference in bacterium (P=0.43). CMV infection (P=0.44), urinary tract infection (P=0.92), and macular edema (P=0.26).Conclusions The present study showed that FTY720(5.0mg)+RDC caused more first treated biopsy-proven acute rejection at twelve months, so FTY720(5.0mg) did not support the reduced-dose cyclosporine A. There was a lower viral infection but higher incidence of bradycardia in FTY720 treated patients. Optimal drug choice may vary between patients.