Objective This study was conducted to investigate the clinical outcomes and safety of primary percutaneous coronary intervention(PCI) combined with tirofiban therapy in patients with acute ST segment elevation myocardial infarction(STEMI). Methods Seventy-one consecutive patients with acute STEMI were divided by random number table to primary PCI combined with tirofiban therapy group(Tirofiban group,25 cases) and primary PCI treatment alone group(Control group,46 cases). Left ventricular ejection fraction(LVEF) and major adverse cardiac events rates(MACE) during hospitalization period and at 30 days discharge and 180 days after discharge were compared between the two groups. Results TIMI grade flow was significantly different between the tirofiban group and control group after surgery. The LVEF and MACE were significantly different between two groups during hospitalization period and at 30 days after discharge. The MACE at 180 days followup was relatively reduced and LVEF was relatively improved in tirofiban group, but there was not significantly different. Conclusion Adjunctive therapy with tirofiban for patients with acute STEMI who underwent primary PCI was safe and effective.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells. CT358 gene from the Chlamydia trachomatis (C. trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedCI vectors. The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins. The GST-CT358 fusion protein was used to immunize mice to raise the antibodies, which specifically recognized CT358 without eross-reacting with other unrelated proteins. The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay (IFA). Meanwhile, pDSRedC 1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection. The hypothetical protein CT358 was identified in the inclusion membrane of C. trachomatis-infected cells for the first time,and it was detected as early as 12 h after C. trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle. Cytosolic expression of CT358 via a transgene failed to affect the subsequent ehlamydial infection. These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein, giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells.CT358 gene from the Chlamydia trachomatis(C.trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedC1 vectors.The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins.The GST-CT358 fusion protein was used to immunize mice to raise the antibodies,which specifically recognized CT358 without cross-reacting with other unrelated proteins.The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay(IFA).Meanwhile,pDSRedC1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection.The hypothetical protein CT358 was identified in the inclusion membrane of C.trachomatis-infected cells for the first time,and it was detected as early as 12 h after C.trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle.Cytosolic expression of CT358 via a transgene failed to affect the subsequent chlamydial infection.These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein,giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

Objective To localize and characterize the plasmid protein pORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods The open reading frame encoding for pORF5 protein from the Ct plasmid was amplified and cloned into the pGEX-6p vector. The recombinant plasmid pGEX-pORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the pORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous pORF5 protein and detect the expression pattern in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). At the same time, ELISA was used to determine whether pORF5 plasmid protein was expressed and immunogenic during Ct infection in humans. Results pORF5 was detected a dominant signal in the cytosol of the Chlamydia-infected cells with a pattern similar to that of anti-CPAF. pORF5 also appeared in the RBs and EBs in small quantity. Athough pattern was similarly, pORF5 did not overlap with CPAF. pORF5 protein was strongly recognized antiserum in an ELISA. Conclusion The pORF5 plasmid protein was identified as a secreted protein with good immunogenicity, pORF5 gene was to express the endogenous target protein during human infection.

Objective To observe the clinical efficacy of acupoint thread embedding on the peripheral serum leptin and insulin levels in simple obesity patients, and to further explore the mechanism of acupoint thread embedding in treating simple obesity.Method A total of 120 patients with simple obesity were randomized into a control group, an eletroacupuncture (EA) group and a thread embedding group, 40 cases in each group. The control group was intervened by diet plus exercise intervention, the EA group by EA treatment in addition to the intervention given to the control group and the thread embedding group by acupoint thread embedding treatment in addition to the intervention given to the control group. The levels of blood serum leptin and insulin in the three groups were observed before and after 2 treatment courses and the clinical efficacies were compared among the three groups.Result The total effective rate was 87.5% in the thread embedding group, 85.0% in the EA group, and 47.5% in the control group. The total effective rates in the thread embedding group and EA group were significantly different from the rate in the control group (P<0.05). The levels of the fasting serum leptin and insulin were significantly changed in the thread embedding group and EA group after the treatment (P<0.01). After the treatment, the levels of the fasting serum leptin and insulin in the thread embedding group and EA group were significantly different from those in the control group (P<0.01,P<0.05). The insulin level in the thread embedding group was better than that in the EA group, and the difference was statistically significant (P<0.05).Conclusion Acupoint thread embedding is an effective approach for simple obesity, and it can down-regulate the fasting peripheral serum leptin and insulin levels.

ObjectiveTo purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein.Methods The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale,and protein G purification by affinity chromatography was used to purify 2H4 McAb.ELISA was used to determine the antibody titer,and identify McAb isotype.Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity.Results The purity of 2H4 antibody was 93％,the titer reached 1:1024,and 2H4 McAb was identified to belong to IgG2a isotype,2H4 McAb reacted strongly with the GST-pORF5 fusion protein and endogenous pORF5 protein expressed by Chlamydia trachomatis serovar A,D,L2,Chlamydia muridarum ( MoPn ),Chlamydia psittaci 6BC,but not other chlamydial plasmid proteins and Chlamydia pneumoniae(Cpn) AR39 strain.Conclusion2H4 McAb against pORF5 protein was successfully purified with a high titer and specificity which lay a foundation for further study on pORF5 protein structure and function.

Objective:To observe the changes of metabolic intensity value of points on the face before and after needling bilateral Hegu (LI 4) in healthy people and provide scientific basis for association between Hegu (LI 4) and face/mouth. Methods:A total of 45 healthy college students were selected in this study. Using medical thermography and Pennes bio-heat transfer model, the infrared thermograph images on the face before and after needling bilateral Hegu (LI 4) were collected to observe the distribution of metabolic intensity value on the face before acupuncture and changes in these values after needling bilateral Hegu (LI 4). Results:Before acupuncture, Cuanzhu (BL 2) had the maximal metabolic intensity value. Its mean value was (0.71±0.23) W. Quanliao (SI 18) had the minimal metabolic intensity value. There were no left-right statistical significances in metabolic intensity values. After needling bilateral Hegu (LI 4), the metabolic intensity values of most points on the face were increased. Kouheliao (LI 19) obtained the maximal increase: 0.35 W on average; and Yangbai (GB 14) obtained the minimal increase: 0.08 W on average. Conclusion:Points on both sides in healthy people have good symmetry in metabolic intensity value. After needling bilateral Hegu (LI 4), the metabolic intensity values of points on the face were increased, especially points around the lips, which accords with the pathway of the Large Intestine Meridian on the head and face. This provided some scientific foundation for the association between Hegu (LI 4) and face/mouth.

Objective To summarize the clinical experience of successful intervention in single chronic coronary actery total ocdusion (CTO) lesions by the transradial.Methods A retrospective analysis was conducted in 103 patients with single CTO lesions who got intervention treatment by the radial artery.Results ( 1 ) Of the 103 cases,57 cases had unstable angina,12 cases had stable angina,and 34 cases chronic myocardial infarction.Lesions' block time was ≤ 6 months in 83 cases,and ＞ 6 months in 20 cases.(2)The path vessels of the 103 patients have no severe tortuosity and anatomical structure variation.Fifty-one cases occurred left anterior descending occlusion,25 cases occurred left circumflex branches occlusion,and 27 cases occurred right coronary artery occlusion.Furthermore,24 cases had chronic complete occlusion,and 79 cases had chronic functional block.The side branches did not block in 91 cases,no lesions(bridge) collateral formation occurred in 87 cases,lesions length was less than 15 mm in 67 cases,and tapered lesions was observed in 81 cases.( 3 ) Final intervention rate via Judkins,XB,EBU guide catheter was 37.86％,30.10％ and 29.13％ respectively.(4)the PILOT successfully through the lesions for the series wire guided was 64.08％.(5) 1.25 mm diameter series with a balloon through the first lesions and successful expanding was observed in 57 cases (55.34％),and 1.5 mm diameter series with a balloon occurred in 38 cases(36.89％ ).Conclusion Intervention treatment by the radial of single CTO lesions is feasible for experienced performers.The successful intervention depends on path vessels unimpeded,target vessels with characteristic pathological features and reasonable choice of instruments.

Objective:To analyze the changes in biological characteristics including infectivity, growth and pathogenicity of Chlamydia muridarum ( Cm) after serial passage in vitro in special conditions in order to provide reference for screening attenuated live vaccines and virulence-related genes. Methods:Wild-type Cm strain (G0) was cultured for several passages using conventional cell culture method under alternate unassisted and assisted culture conditions. Then, the 28th generation (G28) of Cm was selected and compared with the parental G0 strain in terms of centrifugation dependence, attaching ability, intracellular growth curve, plaque size and fallopian tube lesions after genital tract infection in a mouse model. Results:Compared with the parental G0 strain, the G28 strain showed significantly decreased dependence on centrifugation during cell infection ( P<0.05) and increased attachment capacity to cells ( P<0.05). No significant differences were observed in the growth curves 32 h after cell infection or in the plaque sizes between the parental G0 and G28 strains. In the in vivo virulence test, fallopian tube lesions were observed in 87.5% of G0-infected mice and 37.5% of G28-infected mice ( P<0.05). Conclusions:Compared with the parental G0 strain, the G28 strain showed significantly enhanced in vitro infection ability, but decreased in vivo pathogenicity, which brought hope for further identification of virulence genes, isolation of attenuated strains with single genotype and development of live attenuated Chlamydia vaccines.

Objective To investigate the effects of leptin on Treg cells and the possible mecha-nism. Methods Leptin-deficient ( ob/ob) mice and homologous wild-type mice were used in this study. The percentages of Treg cells in spleen tissues and peripheral blood samples were measured by flow cytometry ( FCM) . Differences in Treg cell functionality were compared between the two groups. Splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by magnetic beads, were respectively cultured with leptin and anti-leptin neutralization antibody to evaluate the effects of leptin on Treg cells. Quantitative real-time PCR was performed to analyze the expression of Treg cell-related cytokines at transcriptional level. The levels of IL-10 and TGF-β in the supernatants of CD4+T cell culture were measured with Luminex technolo-gy. Results Compared with the wild-type mice, the ob/ob mice showed higher percentages of Treg cells in both peripheral blood samples and spleen tissues [(11. 56 ± 0. 72)％ vs (5. 47 ± 0. 81)％, (10. 16 ± 0.93)％ vs (6.29±0. 69)％]. Treg cells isolated from the ob/ob mice had stronger immunosuppressive effects on the proliferation of effector T ( Teff) cells and the secretion of TNF-α and IFN-γ than those from the wild-type mice [TNF-α:(1. 6±0. 2)％ vs (2. 4±0. 5)％, IFN-γ:(4. 3±0. 3)％ vs (7. 2±1. 2)％]. The percentages of Treg cells were decreased from (12. 2±1. 8)％ to (7. 6±0. 9)％ upon the in vitro treat-ment of CD4+ T cells from the ob/ob mice with leptin and the immunosuppressive effects of Treg cells were also weakened. However, the percentages of Treg cells were increased from (7. 8±0. 85)％ to (13. 1± 1. 5)％ upon the in vitro treatment of CD4+T cells from the wild-type mice with anti-leptin antibody and the immunosuppressive effects of Treg cells were improved as well. Moreover, the expression of Foxp3, IL-10 and TGF-β at transcriptional level and the levels of IL-10 and TGF-β in the ob/ob group were higher than those in the wild-type group. Conclusions Leptin deficiency significantly promoted the generation of Treg cells in mice and resulted in an increased expression of Foxp3, IL-10 and TGF-βat mRNA level and elevat-ed levels of IL-10 and TGF-β. The treatment of CD4+T cells with leptin might inhibit the generation of Treg cells through down-regulating the transcription of Foxp3, IL-10 and TGF-β.