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 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 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 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.

Objectives To investigate the expression and diagnostic value of 14-3-3 protein in brains of patients with sporadic Creutzfeldt-Jakob disease(sCJD).Methods 14-3-3 protein was immunohistochemically analyzed in tissue from the frontal lobe of 5 patients with sCJD and 4 non-CJD eases Using 14-3-3 ?and ?antibodies with reference to the results of KB,GFAP and PrP detection.Results The expressions of 14-3-3 protein in five brains of sCJD were more obviously,mostly in gray matters and astrocytes in three cases.The concentration was related to PrP deposition type,but not related to prion protein genotype.Except few expression of 14-3-3 protein in neurous of two cases of acute contusion,there were no expression in the other two cases in control group.Conclusions The expression of 14-3-3 protein in brain is useful to pathological diagnosis of CJD.

Objective To analyze negative lymph nodes of 34 non-small cell lung cancer(NCLC) patients with total correction by means of fluorescent quantitation PCR and immunohistcchemistry,and to form molecular bi- ology staging.Methods Clinical data and tissue samples of 193 lymph nodes were collected from 34 patients under- going resection for non-small cell lung cancer.Using fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry method,lymph nodes were examined for CEA gene mRNA,P53 and CK to form molecular biology staging.All the patients were followed-up for an average of forty months.Results The CEAmRNA was identified in 21.7% (42/193) lymph nodes negative patients from 17 patients(17/34,50%); TMN staging was up-regulated in 8 patients;positive lymph nodes were increased in 9 patients.P53 and AE1/AE3 were identified 9.8%(19/193) from 11 patients,18.6 % (36/193)from 15 patients,separately;TMN staging was up-regulated in 2 patients of P53 examination and 5 patients of AE1/AE3 analysis;positive lymph nodes were in- creased in in 7 patients of P53 examination and 11 patients of AE1/AE3 analysis.There was obvious statistical sig- nificance in them,but the molecular biology staging based on the three markers was not an independent factor on re- currence and metasis of lung cancer.Conclusion CEAmRNA.P53 and AE1/AE3 analysis could find lung cancer micrometasis more sensitively to form molecular biology staging which was relative to the prognosis,but not an inde- pendent prognostic indicator.It might be good to the therapy strategy after operation.

Objective To evaluate the value of stereotactic core needle biopsy(SCNB)in diagnosis of breast lesions.Methods Forty-seven cases were punctured with computer-assisted stereotactic system, spring-loaded biopsy guns and 16 G core needles.The record of each item was collected,including clinical manifestations,descriptions of the mammographic characteristics(such as calcification,mass and architectural distortion),the pathology of the SCNB and the surgical pathology or mammographic follow-up data.Then the results of SCNB were analyzed based on the comparison of SCNB pathology and the surgical pathology.The reason that SCNB failed and misdioagnosed was inferred from the relationship of SCNB accuracy and the X-ray characteristics.Results Forty-four cases were punctured successfully,3 cases failed.Thirty-one patients were operated soon after biopsy.The results of 27 SCNB cases agreed well with the final pathology but the other 4 did not.Conclusions SCNB as an accurate,time-saving and cost- effective method,is also minimally invasive and hardly changes the architecture of the breast.SCNB can diagnose breast lesions in advance,reduces the number of surgical biopsy,and is promising in clinical application.

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.