Objective: To identify and highly express the L-Methionine ?-lyase gene from Trichomonas vaginalis(TVMGL).Methods: The TVMGL gene was cloned into pGEX 4T-2,the recombinant protein expressed in E.coli DH5?and confirmed by SDS-PAGE.Results: Denaturing SDS-PAGE analysis confirmed the predicted size of the fusion protein(GST-TVMGL,68 000) and showed high purity after subsequent affinity chromatography on Glutathione sepharose 4B.The target proteins amounted to 34% of the total bacteria proteins.Conclusion: The L-Methionine ?-lyase gene was highly expressed in E.coli.

Objective:To evaluate the biocompatibility Abstract of a liquid of propolis. Methods:According to the relative Chinese national standards,the cytotoxicity of the propolis abstract was investigated by in vitro cell culture,agar diffusion test and the oral mucous membrane irritation response test in rabbits. Results:In the in vitro tests the propolis abstract showed grade 0 cytotoxicity. In the in vivo test the abstract did not result in any pathological change in oral mucous membrane.Conclusion:The propolis abstract is biocompatible.

Objective Clinical investigation of treatment of osteoporotic vertebral compression fractures by unilateral kyphoplasty.Methods After Surgical intervention of sixteen vertebral compression fractures by unilateral kyphoplasty,then investigate the changes of height and Cobb angle of vertebral bodies.Results All surgical procedures were finished safely.The mean height of the anterior and media vertebral bodies was 1.8?0.3cm,1.4?0.4cm preoperatively and 2.2?0.4cm、2.3?0.3cm postoperatively;the Cobb angle was decreased from 28.4??10.2? preoperatively to 19.2??4.5? postoperatively.The mean absolute value of the difference in height between right and left side of the vertebral bodies was 0.1cm in this group.Conclusions The kyphosis is improved and the operative time is reduced by unilateral kyphoplasty.Though this result is satisfied,more clinical studies are needed to be done for authenticating the effect of unilateral kyphoplasty on the treatment of vertebral compression fractures in the future.

OBJECTIVE:To improve the proposal put forward by a hospital on the pharmacoeconomic evaluation of community-acquired pneumonia.METHODS:The domestic pharmacoeconomic evaluation papers on the community-acquired pneumonia were evaluated.The problems lie in the proposal put forward by a hospital on the pharmacoeconomic evaluation of community-acquired pneumonia were analyzed and the measures for its improvement were put forward.RESULTS:The proposal had problems in research angle,determination of sample size,cost calculation,determination of quality of life etc.CONCLUSION:Measures for the improvement of the proposal should be put forward from the aspects of model construction,sample size determination,the design of quality of life table and sensitivity analysis.

Objective To explore the effect of estrogen on the permeability of retinal blood vessel by ovariectomy. Methods Twenty-two healthy rats were divided into experimental and control group randomly. Estrogen level of rats decreased due to ovariectomy in the experimental group while stabilized by sham-ovariectomy in the control group. The results were confirmed by vaginal epithelium smearing. Retinal vein occlusion was established by photodynamic method, and leakage of Evan's blue in retina was determined by spectrophotometer. Results Mature value of vaginal epithelium decreased significantly in ovariectomy rats( t=21.008,P =0.000) while not significantly in sham-ovariectomy ones ( t=0.319,P =0.756); the mean leakage of Evans blue was (25.503 0?4.378 47) ng/mg in experimental group, and (17.830 0?4.265 69) ng/mg in the control group, and the difference between the two groups is significant( t=3.969 36,P =0.001). Conclusion Ovariectomy is an useful method to study the effect of estrogen on ocular diseases, and when estrogen level decreases, the permeability of retinal blood vessel increases.

Objective To investigate the effects of demethylating agent 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the biological behaviors and the expression of RASSF1A of lung carcinoma cells H460. Methods Lung carcinoma cell line H460 were treated with 5-Aza-CdR. Cell proliferation was determined by MTT assay and colony forming test. The methylation status of RASSF1A gene was detected by PCR. The expression of RASSF1A protein was measured by Western blotting, and the cell cycle was analyzed by flow cytometry. Results With 5-Aza-CdR treatment, the proliferation speed of H460 lung carcinoma cells was slowed down and the colony formation rate of H460 cells was decreased significantly compared with control group (38.5 %, 27.5 % and 60.5 % in 5-Aza-CdR 5 and 10 μmol/L groups and control group, respectively, P <0.05). The methylation degree in the promoter of RASSF1A gene was decreased and the expression of RASSF1A protein was detected after 5-Aza-CdR treatment. 5-Aza-CdR induced G1 phase arrest of the H460 cells. Conclusion The hypermethylation of CpG island in the promoter of RASSF1A gene results in the loss of RASSF1A protein expression in human Lung carcinoma cell line. The demethylating agent 5-Aza-CdR could restore RASSF1A gene expression.These findings provide theoretic evidence for clinical treatment of human lung carcinoma with demethylation agent 5-Aza-CdR.

Objective To study the effect of allicin on the growth and radiosensitivity of human pancreatic carcinoma BXPC3 cells.Methods BXPC3 cells were exposed to X-rays in the presence or absence of allicin.Cell proliferation was measured by MTT assay.Cell cycle distribution and apoptosis were detected by flow cytometry assay.Cell radiosensitivity and the influence of allicin on it was evaluated by colony formation assay.The expressions of Bax and Bcl-2 proteins were assayed by RT-PCR and Western blot.Results IC50 values of allicin on cell growth were 76.24,58.34 and 43.58 μmol/L under 12,24 and 48 h treatment,respectively.Treatment of cells with allicin obviously inhibited cell growth after irradiation and hence increased radiosensitivity (t =2.74,P ＜ 0.05).This treament also enhanced radiation-induced cell cycle arrest at G2/M phase (t =11.41,P ＜0.05),apoptosis induction (t =12.36,P ＜ 0.05),and Bax expression (t =4.83,P ＜ 0.05),but it decreased Bcl-2 expression (t =3.69,P ＜ 0.05).Conclusions Allicin could inhibit cell growth,induce cell cycle arrest and apoptosis via Bax/Bcl-2 pathway and hence increases radiosensitivity of BXPC3 cells.

Objective To investigate the value of the cold supernatant in the clinical practice. Methods 25 cases of TTP got a direct infusion of cold supernatant or plasma exchange using cold supernatant as fluid replacement and with medications at the same time. Then we observed the clinical treatment effect of all the cases. Results The clinical remission rate of 25 cases of TTP was 80%. Conclusion The cold supernatant could be used as a new blood component in clinical application,especially for the treatment of thrombotic microangiopathy (TMA). It not only can save valuable blood resources,but also can reduce the economic costs of patients.

Objective To investigate the mechanism and prevention of gallstones after esophageal cancer surgery.Methods Clinical data from 297 cases of esophageal cancer patients underwent surgical treatment from 2007 to 2010 inour hospital were retrospectively analyzed for the incidence of post-operative gallstones.ResultsNo cholecystitis and gallstones were detected by B-ultrosound before the operation in the 39 patients.13.1％ of the patients developed gallstones from 1 month to 3 years after the operation.The incidence was significantly higher than that in general population.Conclusion There is a high chance of gallstones in patients after radical resection of esophageal cancer.Cutting vagus nerve plays an important role in the incidence of gallstones.Preservation of vagus nerve during the surgery should be performed as possible.

Purpose:To evaluate the effect of late accelerated hyperfractionated radiotherapy (LCAHFR) combined with chemotherapy on advanced esophageal cancer.Methods:176 cases of advanced esophageal cancer were treated from March 1994 to March 1997.Patient were divided into 2 groups, including radiotherapy alone group (80 cases) receiving conventional irradiation to a total of 64～70 Gy and combined treatment group (96 cases) which received LCAHFR groups plus chemotherapy with PF regimen. Combined treatment group patients were treated with conventional fractionated radiotherapy during the firs two thirds of the treatment to a dose about 40 Gy/20 F/4 weeks. This was followed by accelerated hyper fractioned radiotherapy using reduced fields, twice daily at 1.5 Gy per fraction to a dose about 24～30 Gy.All cases of the combined treatment group were treated with PF(DDP 20 mg iv drip on days 1 to 5,and 5 FU 750 mg iv drip on days 1 to 5,Every 28 days as one cycle).All patients were followed for ≥ 5 years.Results:The short term results of LCAHFR combined with chemotherapy was better than that of radiotherapy alone. The difference between these two groups was statistically significant( P 0.05).The one , three and five year survival rate were 69.8%,51.1%,29.2% in combined group and 46.3%,21.3%,11.3% in simple irradiation alone group. The survival rates in the combined group were much better than those in simple radiotherapy group( P