Objective To explore feasibility and clinic results of the MIPPO (minimally invasive percutaneous plate osteosynthesis) technique in treatment of humeral fractures. Methods 14 patients with humeral fracture underwent the MIPPO operation in our department. 7 of them were injured in a traffic accident, 4 in sports and 3 in daily life. There were 11 males and 3 females, with their ages ranging from 16 to 72 years. Of the 14 cases, 9 were proximal ones and 5 mid- distal ones, 3 were multiple injuries and 1 pathological fracture. Results The average intra- operative time was 80 minutes (ranging from 55 to 130 minutes). No patient received blood transfusion. The intra- operative blood loss was 100 to 200 mL. 14 cases were followed up for 8 weeks to 14 months. Their cuts healed at the primary stage. Their shoulder and elbow functions were fine. The fracture segments got satisfactory reduction with good apposition and alignment radiologically. The callus appeared 4 to 8 weeks postoperatively. Conclusion MIPPO is a safe and effective treatment for the humeral fracture with the benefits of less invasion, fewer complications and higher union rate.

Objective To clone and study the antimicrobial-resistant gene in Neisseria gonorrhoeae.Methods The gene library,which contains the differential genes of antimicrobial resistant strains and stan-dard reference strains of Neisseria gonorrhoeae,was constructed using a technique known as suppression subtractive hybridization(SSH).Then the antimicrobial resistance associated genes were cloned and ana-lyzed.Results Subtractive gene library in antimicrobial-resistant Neisseria gonorrhoeae was successfully constructed,which contains2500positive clones.Sequence analysis was performed on5clones.The se-quences of these five clones were unknown previously.Conclusions The subtractive DNA library is succes-sively constructed which may provide an important clue for studying the mechanism of antimicrobial resis-tance in Neisseria gonorrhoeae.

Objective:To re-establish the quality control standard for Weiling granules. Methods:The 4 chief herbs in the prepa-ration, radices paeoniae alba, licorice, rhizoma corydalis and hawthorn were identified by TLC qualitatively. The content of paeoniflor-in in radices paeoniae alba was determined by HPLC. The separation was performed on an Agilent XDB-C18 (250 mm × 4. 6 mm, 5μm)coulme with mobile phase consisting of acetonitrile-0. 1% phosphoric acid solution (10:90). The detection wavelength was 230 nm and the flow rate was 1. 0 ml·min-1 . Results:The spots in TLC were clear without any interference. The linear range for paeoni-florin was 0. 151-1. 212 μg(r=0. 999 9,n=5). The average recovery was 99. 63% and RSD was 2. 01%(n=9). Conclusion:The method is simple and accurate with high reproducibility, which can be used for the quality control of Weiling granules.

Objective:To analyze the frequencies of HLA-A*0201 restricted CEA-specific CD8+T cells, HLA-A*0201/FLUmp tetramer and HLA-A*0201/CAP-1 tetramer were applied in patients with colorectal cancer.Methods: Lymphocytes from peripheral blood and lymph node,1×106 cells/ml,were incubated with 1μg HLA-A*0201/peptide tetramers and anti-CD8 for 1 h at 25 coseperately.The cells were then washed in PBS.Next,the cells were illuminated by detecting frequencies of FLUmp-specific CD8+T cells and CAP-1-specific CD8+T cells with flow cytometry.Results: HLA-A*0201/peptide were used to detect CAP-1 or FLUmp-specific CD8+T cells,which were analyzed either healthy individuals or patients with colorectal cancer.We did not find differences in average frequencies of FLUmp-specific CD8+T cells between 11 HLA-A*0201+patients with colorectal cancer and 14 HLA-A*0201+healthy individuals [ ( 0.671 ±0.421 )%, ( 0.564 ±0.408 )%].But the frequencies of CAP-1-specific CD8+T cells of HLA-A*0201+patients with colorectal cancer showed higher than HLA-A*0201+healthy individuals [ ( 2.409 ± 2.385 )%, ( 0.020 ± 0.021)%respectively],which was statistically significant(P=0.008).Conclusion:The frequencies of CAP-1-specific CD8+T cells in PBMC from peripheral blood and lymph node of HLA-A*0201+patients were increased,showed CEA-specific CTs has a vital role in colorectal cancer.

Objective To study the role of human carcinoembryonic antigen-related cellular adhesion molecule 1 (hCEACAM1)in mediating the specific adhesion of N. gonorrhoeae to its human host cells.Methods A recombinant eukaryotic expression vector pCDPGICEA1 was constructed by putting hCEACAM1 cDNA behind both hCD46 promoter and rabbit β-globulin intron 2,and with which,the COS-1 cells were transfected. Following G418 selection, the COS-1 cells expressing hCEACAM1 were sorted out with flow cytometry. The adhesion of N. gonorrhoeae to the gene transfected COS-1 cells was analyzed with bacterial binding assay. Results hCEACAM1 cDNA could be expressed effectively under the direction of hCD46 promoter and rabbit β-globulin intron 2,and N. gonorrhoeae could adhere to COS-1 cells expressing hCEACAM1. Conclusion hCEACAM1 can mediate the adhesion of N. gonorrhoeae to animal originated COS-1 cells. thus its transgenic mice may be used as a novel animal model for studying N. gonorrhoeae infection.

Objective:To illuminate the influence of InsB15-23 H-2Kd dtSCT to the morbidity of type 1 diabetes mellitus in NOD mice.Methods:An eukaryotic plasmid encoded membrane-expressed InsB15-23 H-2Kd dtSCT was inoculated into 3 weeks old female NOD mice subcutaneously and the blood sugar and morbidity of type 1 diabetes mellitus were monitored once a week.To illuminate the cellular mechanism of immunologic intervention of membrane-expressed InsB15-23 H-2Kd dtSCT to the course of type 1 diabetes mellitus in NOD mice,the mononuclear cell infiltration of islets was detected by tissue slice and the frequency of IGRP206 2-14 specific CTLs in PBMC was analyzed by FACs.Results: As compared with pcDNA3.1 (-) control ( 60%) and untreated NOD mice ( 80%) , mice immunized with InsB15-23 H-2Kd dtSCT exhibited low level of islet infiltration and low morbidity in 30 weeks old ( 9%) .But the frequency of IGRP206-214 specific CTLs in PBMC of 16 and 40 weeks old mice showed no difference.Conclusion:Membrane-expressed InsB15-23 H-2Kd dtSCT can protect NOD mice from type 1 diabetes mellitus in IGRP206-214 independent pattern.

Objective To establish a mouse model of bacterial vaginosis ( BV) by infecting estro-gen-treated mice with Prevotella bivia ( P. bivia) . Methods The mice were intraperitoneally injected with beta-estradiol 17 valerate which was suspended in sesame oil and then inoculated with different doses of P. bivia strains at the logarithmic phase. Samples of vaginal flushing fluid were collected at different time points after inoculation and used for the isolation of P. bivia strains and the detection of sialidase activities. Altogether 30 mice treated with estrogen and high dose of P. bivia were killed on days 2, 7 and 21 (n=10). Samples of cornua uteri, bladder and kidney were collected from those mice for P. bivia strains isolation. Re-sults Injection the estrogen-treated mice with P. bivia via vagina could cause the P. bivia infection for more than 14 days. The numbers of P. bivia strains isolated on day 21 decreased significantly. Enhanced sialidase activities and clue cells were observed in vaginal secretions of mice with P. bivia infection. Injection of mice with the high dose of P. bivia could spread the infection to cornua uteri. Conclusion Estrogen-treated mice could be used as an animal model for researches on BV.

Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.

AIM: To study the effects of recombinant soluble MHC class Ⅰ chain-related protein A (sMICA) on the cytotoxicity, secretion of IFN-γ, proliferation and apoptosis of peripheral NK cells. METHODS: After NK cells were cocultured with recombinant soluble MICA proteins overnight, the cytotoxicity of NK cell on target cells was detected by flow cytometry. The supemant was collected to determine the concentration of IFN-γ by ELISA. The proliferation of NK cells to sMICA was detected by MTS/PMS. NK cells were labeled with annexin V and PI to analyze their apoptosis. RESULTS: Soluble MICA inhibited the cytotoxicity of NK cells and down-regulated the secretion of IFN-γ, but it showed no effects on the proliferation and apoptosis of freshly isolated peripheral NK cells. CONCLUSION: The soluble MICA shedding from tumor cells could be a pathway of cancer immune evasion by down-regulating the biologic activities of NK cells.

Objective To develop fast detection techniques for the diagnosis of gonococcal infections.Methods Prokaryotic expression vector for Neisserial surface protein A (NspA) was constructed using the NspA gene cloned by PCR.Mice were immunized with the renatured recombinant NspA (rrNspA)to prepare antibodies against NspA.Western blot and ELISA was used to analyze the binding of NspA antibodies to lysate of gonococcal cells or to intact gonococci.Results NspA antibodies that were prepared by the rrNspA expressed in E.coli could bind to rrNspA,the natural NspA existing in gonococcal cells,or intact gonococci.Conclusion RrNspA and its antibodies have potential value in developing fast diagnostic kits for gonococcal diseases.