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.

ObjectiveTo examine the relationship between HIF-1αt and VEGF expression and the clinicopathological characteristics in hypopharyngeal squamous cell carcinoma. Methods The expression status of HIF-1α and VEGF were examined by immunohistochemistric method (IHC) in 62 tumor tissue and 19 paracancerous normal tissue.The relationships between the expression of HIF-1α and VEGF and clinicopathological characteristic were analyzed. Results HIF-1αt and VEGF expression were higher in hypopharyngeal carcinoma tissues than those in paracancerous normal tissues (66. 1％:26. 3％ ;x2 =18. 02,P ＜0. 05 ;67. 7％ : 31.6％ ; x2 =19.22, P ＜ 0. 05 ). The expression intensity of HIF-1 α was related to T stage, N stage and TNM stage ( x2 =4. 23,5.83,9.94,all P ＜0. 05). The expression intensity of VEGF was related to metastasis, T stage, N stage and TNM stage (x2 =5.62,7. 38,15.75,4. 29 ,all P ＜0. 05 ). There was minus relationship between overall survival and expression level of HIF-1 α and/or VEGF (x2 =29. 25, P＜0.01; x2 =24.88, P＜ 0.01 ).On multivariate analysis,HIF-1α expression and T stage were independent prognostic factors for overall survival (x2 =4.80,5.74, all P＜0. 05).ConclusionsHIF-1α and VEGF may be considered as a parameter in evaluation of progression, metastasis and prognosis of hypopharyngeal carcinoma and also may be a direction of molecular target therapy.

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 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.

Objective To observe the effect of reperfusion therapy on the prognosis of acute myocardial infarction (AMI) complicated by eardiogenic shock(CS)in reperfusion era.Methods 89 cases of AMI with CS were included with 57 male and 32 female.50 cases received conservative therapy and 39 cases reperfusion therapy.28 of the 39 cases had suecessflll reperfusion and 11 eases failed.18 patients had intra-aortic balloon pump (IABP) within 1 hour of CS,they constituted an early group;35 patients treated with IABP 1 hour after CS were of a late group.A group of 36 cases were not treated with IABP (no IABP group).Results The mortality of the early group with IABP Was significantly lower than that of the late and no IABP group(33.3% vs.74.2% vs.86.1%,P＜0.01).The mortality of the group with sucessful reperfusion was significantly lower than that of unsuccessful reperfusion and conservative no IABP group (42.8% vs.81.8% vs.84.0%,P＜0.01).logistic regression analysis showed that successful reperfusion therapy (OR 4.232,95% CI 1.407～12.730,P=0.01) and THE TIME of using IABP(OR 0.22.95% CI 0.063～0.764,P=0.017)were independent risk factors for death.Conclusion Early successful reperfusion and early institution of IABP were the most important therapeutic measures for reducing mortaliIv of AMI complicated by CS.

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 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 understand the characteristics of schistosome cercarial infested water contact of residents in Poyang Lake endemic areas after the operation of Three Gorges Project,so as to provide the evidence for formulating a well-directed con-trol strategy. Methods The residents in two villages in schistosomiasis endemic areas were selected and surveyed for their activi-ties of infested water contacts by questionnaire. Results The proportion and frequency of infested water contacts of residents in Poyang Lake region were high and the average rate of infested water contacts was 69.77%,and 98.96%of infested water contacts occurred in spring,summer and autumn. The main ways of infested water contacts were fishing/swimming/playing for adult men, washing clothes/fishing for adult women,and swimming/playing for school children,respectively. The rates of infested water con-tacts were 73.33%,71.63%and 61.87%in adult men,women and school children,respectively. The frequencies of infested water contacts>1 time/week,1 time/week-1 time/month,and<1 time/month were 62.61%,23.74%and 13.66%respectively. Conclu-sions The infested water contacts mainly occurs in spring,summer and autumn,and the main way of infested water contacts was fishing. Farming and mowing grass,which had been the primary way,now become the secondary way for the adult residents.