Objective To clone and express the new gene R049 of uropathogenic Escherichia coli 132.and to investigate the immunopmtective effects of the R049 recombinant protein on mice.Methods The pmkaryotic expression system of gene R049 was constructed by directed cloning.Thereafter,the R049 recombinant protein Was expressed and purified by Ni affinity chromatography.Polyclonal antibody was pre-pared by immunizing BALB/c mice with R049 recombinant protein.The R049 recombinant protein and whole bacterial proteins of UPEC132 were analyzed by SDS-PAGE and Western blot.BALB/c mice were im-munized with R049 recombinant protein before challenged by UPECl32 through urinary tract.Then the differences of urine and renal colony counts between immunization group and control group were compared.Results The recombinant strain E coli BL21(DE3)/pET32a-R049 ORF was constructed successfully,and the relative molecular mass of the R049 recombinant protein was 66.9×103 and its purity was up to 95% af-ter purification.The titer of polyclonal antibody wag≥1:102 400 analyzed by indirect ELISA.Both of the R049 recombinant protein and whole bacterial proteins of UPECl 32 were confirmed to show specffic reactions on the antiserunl throughh Western blot.The animal experiments showed the urine and renal colony counts of immunization group were significantly lower than that of the control group(P<0.01,P<0.05).Conclu-sion The new gene R049 of uropathogenic E.coli 132 had immunopmtective effects on mice and the defini-tive mechanism would be needed to further study.

Objective To observe the effect of salmeterol/flutieasone propionate on children with asthma. Methods Salmeterol/flutieasone propionate inhalation of different dosages were given according to different illness condition of asthma. Investigate the effects of 180 days inhalation of SM/FP among children. Results After 180 days' treatment, the PEFam and PEFpm of those children of asthma were significantly increased. The lung function has a perceptible improvement. In addition, the daytime and noctural symptom scores and asthma symptom were improved. Moreover, the shore-acting β2 agonist was used with decreasing frequency. The effects of SM/FP inhalation were marvelous after 1 week of treatment and should make a long persisting time alter six months of treatment. Conclusion SM/FP is an effective medicine for children with asthma and should be make more widespread.

Objective To select more rapid,sensitive and specific method in detection of respiratory syncytial virus(RSV)directly from clinical specimens.Methods RSV was detected by virus isolation in tissue culture,direct smears and detection by indirect immunofluorecence assay(IFA),rapid culture assay,sandwich enzyme linked immunosorbent assay(ELISA)as well as labbed streptavidin biotin method(LSAB)from 45 specimens(nasopharyngeal aspirates,NPAs) collected from infants and young children with acute lower respiratory tract infection.Results Of 45 NPAs,12 cases(26.7%) were positive by virus isolation,14 cases(31.1%) were positive for RSV by direct detection of RSV antigen by IFA,20 cases(44.4%) were positive with rapid culture assay,4 cases(8.9%)were positive by sandwich ELISA,4 cases(8.9%)were positive by LSAB.Conclusion Rapid culture assay and direct detection of RSV in NPAs direct smears by IFA are rapid,sensitive method in the diagnosis of RSV infections.

Objective Gambogic acid ( GA) can suppress the growth of multiple tumor cells , including gastric carcinoma , hepatoma , hematologic neoplasms and breast carcinoma , but there have been few reports about its effect on urologic neoplasms .This study was to investigate the possible mechanisms of GA inducing bladder cancer cell apoptosis and cell cycle arrest . Methods We cultured human bladder cancer BIU8-7 cell lines in vitor and treated the cells in the logarithmic growth phase with isotonic saline solu-tion (negative control)or GA at the concentrations of 1.0, 2.0, and 3.0μmol/L, respectively.We determined the expression of the Caspase-3 protein in the tumor tissue using the immunohistochemical S-P method and detected GA-induced apoptosis of the bladder cancer cells and cell cycle changes by flow cytometry . Results The expressions of the Caspase-3 protein were 4.28 ±1.86, 5.03 ± 0.78, and 6.47 ±1.31 in the 1.0, 2.0, and 3.0μmol/L GA groups, respectively, significantly higher than 2.13 ±1.27 in the nega-tive control (P<0.05).Flow cytometry showed a gradual decrease of the cells in the G 0/G1 phase and a gradual increase in the G2/M phase , but no obvious change in the S phase . Conclusion Gambogic acid can promote the apoptosis , arrest the cell cycle , and in-hibit the proliferation of bladder cancer cells by increasing the expression of the Caspase -3 protein.

Objective To study the recogniting patients identity for the safety and reliability of radiotherapy. Methods Through PDCA 4 footwork, namely, plan, do, check, action the technicians in the hospital to improve patients' identity verification.Results After 4 months of PDCA cycle,the patient identity verification qualified rate increase gradually,from 88.17% up to 99.07%,the privacy of patients satisfaction rate rose from 52. 69% to 98. 15%. The patients identification accuracy rate of 100%, technicians working efficiency has been greatly improved. Conclusions The measure of patient identification can improve the working process of radiotherapy for safety and efficiency and can get better privacy protection.

Objective To study the influence of the angle of multileaf collimator leaves on segments (control point for sliding windows) number and monitor units in intensity-modulated radiotherapy (IMRT) plan optimization.Methods 10 cases undergone IMRT with rectangle target volume were choosed,2 plan were designed with step and shot technique and sliding windows technique for running direction of collimator with target long axis vertical and parallel respectively,compared the plans with similar optimization parameters for different angle of the collimator.Results The number of segments and monitor units increased by 52.8％ and 49.6％ more than longitudinal direction of leaf collimator with static IMRT respectively,there appeared significant difference (P =0.000) ; and the number of segments and monitor units increased by 58.2％ and 61.9％ more than longitudinal direction of leaf collimator with dynamic IMRT respectively,there appeared significant difference (P =0.000).Conclusions For the optimization of IMRT based on multileaf collimator,the direction of collimator perpendicular to the target long axis,there could be obviously decrease the number of monitor units,shorten the treatment time,improve the treatment efficiency with similar dose distribution.

Objective To investigate the inhibitory effects of overexpression of PTEN on renal epithelial-mesenchymal trans-differentiation induced by TGF-β1,and the signaling transduction mechanism.Methods HKC cells were transfected with GFP-PTEN via lipofectAMINE2000.The efficiency of transfection was detected by fluorescence microscope.The expression of PTEN protein and mRNA in the translected cells was detected by Western blot and RT-PCR respectively.The experiment was divided into four groups:normal group,TGF-β1 stimulation group,GFP-PTEN+TGF-β1 group and empty vector+TGF-β1 group.The expression of E-cadherin,a-SMA,Akt and p-Akt was detected by Western blot.Results Most ceils transfected with GFP-PTEN expressed GFP.The expression of PTEN protein and mRNA was strongly increased when HKC cells were transfected with GFP-PTEN(all P<0.05).In both TGF-β1 stimulation group and empty vector+TGF-β1 group,the expression level of E-cadherin was lower(all P<0.05),while that of p-Akt and a-SMA was higher than in normal group(both P<0.05).The expression level of p-Akt and a-SMA in GFP-PTEN+TGF-β1 group was Iower(both P<0.05),while that of E-cadherin was higher than in TGF-β1 stimulation group and empty vector+TGF-β1 group(both P<0.05).The expression of Akt was similar in the four groups.Conclusion Overexpression of PTEN can inhibit renal epithelial-mesenehymal trans-differentiation induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.

Objective To analyze safety,efficacy and resection methods of video-assisted thoracic surgery(VATS) for the treatment of intralobar pulmonary sequestration(IPS).Methods Data of 17 patients who were diagnosed as IPS and received VATS from December 2006 to September 2011 were retrospectively analyzed.The patients were 7 males and 10 females with the mean age of 40.3 (14-61) years.Diagnosis was confirmed in 9 patients by enhanced CT and unconfirmed in 8 patients.Three ports were used for surgery.After the aberrant artery was confirmed,liner stapler was used in 16 patients to cut it and Hem-o-lok was used in 1 patient because the aberrant artery was about 3 mm in diameter and long enough.If the diameter of the aberrant artery was longer than 10 mm,a stapling device without knife was used to occlude it centrally and a second stapling device was used to cut it peripherally.Wedge resection or lobectomy was performed due to the different conditions.When the lesion was small with linited range in CT image and the lesion was easily distinguished from normal lung tissue during operation,wedge resection was preferred.Results Seventeen patients underwent VATS successfully without any conversion to thoracotomy or any serious complications.Five patients were planned to receive wedge resection and one was converted to lobectomy.Another 12 patients were planned to receive lobectomy and all succeeded.The mean operating time was 128 (80-170)min.The mean blood loss was 80 (5-200) ml.The mean days of chest tube maintained were 4.0 (2-6) days.The mean postoperative hospitalization days were 7.6 (4-11) days.All patients were diagnosed as IPS according to operating in-sight and postoperative pathology.There was no patient suffering from chronic cough,bloody sputum or recurrent pneumonia during the follow-up.Conclusion VATS for the treatment of IPS is safe and feasible.If conditions permit,wedge resection or segmentectomy may be preferred.

Objective To systematically review the evidence for the effect of vitamin D supplementation on insulin resistance and blood glucose in people who are obese or people with abnormal glucose metabolism. Methods We searched databases including Pubmed, Elsevier, Web of Science, and WANFANG Database etc. for randomized controlled trials comparing vitamin D or analogues with placebo. We extracted data on insulin resistance and blood glucose, including homeostasis model assessment of insulin resistance( HOMA-IR), fasting blood glucose, HbA1C (% ). A1l data were analyzed using Review Manager 5. 0. Results Nine studies involving 867 participants were included. The results of meta-analysis showed that: ( 1 ) For people who are obese and with abnormal glucose metabolism, meta-analysis showed a small improvement in HOMA-IR(SMD -0. 34,95% CI -0. 61 to -0. 06, P<0. 05) and a small effect on fasting glucose (SMD -0. 41 mmol/ L, 95% CI -0. 68 to -0. 15, P<0. 05),while such effects were not seen in people who are obese but with normal blood glucose. (2) No serious adverse events were associated with the administration of vitamin D. Conclusion vitamin D supplementation may be benefit for improving insulin resistance and fasting blood glucose in people who are obese and with abnormal glucose metabolism, but has no effect on obese people with normal blood glucose.

Objective To investigate the serum amyloid A (SAA) and interleukin-18(IL-18)concentration in the pathogenesis of type 2 diabetes mellitus (T2DM) and its macrovascular complications, and study the relation between them. Methods ELISA was used to assay serum SAA and IL-18 levels in 65 T2DM patients (including 31 cases with macrovascular complications) and 30 healthy controls. Results Serum SAA and IL-18 levels [(3.09±0.96)mg/L, (98.8±36.4)ng/L]were significantly elevated in patients with T2DM as compared with those in control subjects [(1.06±0.45)mg/L, (58.9±15.6)ng/L](P<0.05). There was significant difference of SAA and IL-18 levels between T2DM patients with [(6.34±1.52) mg/L,(141.2±48.3)ng/L]and without macrovascukar complications [(2.65±0.39)mg/L, (80.2±20.1)ng/L](P < 0.05).Univariate linear regression analysis showed significant positive correlations between serum IL-18 with SAA (r =0.615, P<0.05), SAA, IL-18 and fasting blood glucose (FBG) had mutual positive correlations (r=0.312, 0.428, P< 0.05, respectively). Conclusions In patients with T2DM, serum SAA and IL-18 concentration is greater than in non-diabetic subjects. SAA and IL-18 play important roles in the initiation and development of T2DM. The study suggests that SAA and IL-18 might be an important independent risk factor.