OBJECTIVETo construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis.

METHODSlipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosis patients.

RESULTSOf 493 acute leptospirosis patients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosis patients with typhoid fever, hemorrhagic fever or dengue fever.

CONCLUSIONTrigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.

Antigens, Bacterial ; genetics ; metabolism ; Bacterial Outer Membrane Proteins ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; immunology ; Leptospirosis ; diagnosis ; metabolism ; Lipoproteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

Objective: To observe the safety and clinical efficacy of tumor antigen-pulsed dendritic cell(DC) vaccine in treatment of advanced malignant tumor.Methods: Ninety-one patients with non-small cell lung cancer,colon and rectal cancer,melanoma,renal carcinoma,breast cancer and other malignant tumors were enrolled in this study.All patients met the selecting standard and signed informed consent.Human dendritic cells were obtained from peripheral blood monocytes by culturing them with granulocyte macrophage-colony stimulating factor and interleukin-4.DC vaccine was prepared from tumor antigen pulsed immature dendritic cells in vitro.Patients received the vaccine therapy once every week and one cycle was defined as once every week for 3 weeks.Results: All the patients received 96 cycles of DC vaccine treatment.Symptoms of toxicity included fever,shivering,aching pain of muscle,asthenia,itching,stifle and transient fatigue;most of the symptoms automatically recovered.Clinical efficacy of the treatment was evaluated in 76 patients.Thirty-one of the 76 patients were stable after treatment and 45 were in progressive situation,with the clinical benefiting rate being 40.8%.Eighty-five patients were followed up.The median time for progression was 2.6 months;the overall survival time was 0.9-30.6 months;and the median survival period was 4.5 months,with the one year survival rate being 9.2%.Conclusion: The results suggest that the DC vaccine therapy is well tolerated in treating patients with advanced malignant tumors and has satisfactory clinical benefit;the clinical value of DC vaccine therapy needs to be further observed.

Objective To investigate the security and efficacy of one stage and bilateral resection of thymic tissue and clearance of mediastinal fat by uniportal-video-assisted thoracic surgery(VATS) to cure patients with myasthenia gravis(MG).Methods A number of 131 patients with MG who underwent resection of thymic tissue and clearance of mediastinal fat by VATS in one single center from February 2009 to December 2013 were selected in this retrospective study.76 patients underwent unilateral resection of thymic tissue and clearance of mediastinal fat by three portal VATS from February 2009 to March 2012 and 55 patients underwent one stage and bilateral resection of thymic tissue and clearance of mediastinal fat by small uniportal-VATS from April 2012 to December 2013.The time for operation,the bleeding volume during operation,the volume of postoperative drainage and drainage time,the improvement of symptoms,the postoperative pain,hospital stays and the occurrence of myasthenia gravis crisis were compared between the two groups.Results The general condition and pathological type did not have significant statistical differences between the two groups.The operating time in the uniportal-VATS group was significantly longer than that in three portal VATS group,but the pain was lighter,and the hospital stay was shorter.There were no significant differences between groups in terms of blood loss,postoperative drainage time,and volume of drainage.The follow-up was from 32 to 90 months,and 118 (90.08％)patients completed the follow up.94.5 ％ of the patients in uniportalVATS group acquired complete stable remission(CSR),while it was 84.2％ in three portal VATS group(P ＜ 0.05),and the uniportal-VATS group had lower rate of myasthenic crisis (P ＜0.05).Conclusion One stage and bilateral resection of thymic tissue and clearance of mediastinal fat by small uniportal-VATS is safe and effective with shorter hospital stay and less pain,and it can get higher CSR and less myasthenic crisis,its efficacy is superior to traditional three portal VATS.