Objective To obtain the heavy chain variable region (VH) gene of monoclonal antibody 2F2 against Japanese encephalitis virus (JEV).Methods Total RNA was isolated with Trizol from hybridoma 2F2 cells,and cDNA of VH was amplified with reverse transcription polymerase chain reaction (RT-PCR) and sequenced.The putative VH gene was expressed in E.coli,and the expressed products was detected with enzyme linked immunosorbent assay (ELISA) to determine the activity to bind JEV.Results The VH gene was 354 bp in length which encodes 118 amino acids.Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the VH gene was successfully expressed and purified from inclusion bodies.ELISA result also demonstrated that VH gene expressed products bind purified JEV.Conclusions The VH gene of monoclonal antibody 2F2 against JEV had been cloned.

Objective To observe stimulation and regulation of T lymphocyte subtypes by wild-type and mutant A?40 and A?42 conjugated with different adjuvants and to investigate possible ways to lower the toxicity of A? immune response. Methods Forty BALB/c mice were randomized into five groups:Al adjuvant group,A?42+CFA group,A?42+Al group,A?40+Al group and A?40(E22A)+Al group. After an initial immunization and booster immunization,antibody was titrated. Their splenocytes were stimulated by respective antigen. After 48 h,the quantities of IFN-?,IL-2,TNF-? and IL-4 were tested. After 72 h of culture,the proliferative response of splenocytes was detected by CCK-8 assay.Results Experimental groups generated specific anti-A? antibodies with proliferative response of spleen lymphocytes. Cytokine detection in culture supernatant showed that A?42+CFA group had the highest secretion of Th1-type cytokines IFN-?,IL-2,TNF-? when A?40(E22A)+Al group had the lowest(P

Objective To investigate the distribution and drug resistance of pathogens isolated from urine culture in 2013 and provide basis for clinical treatment .Methods MicroScan WalkAway 96 PLUS bacterial identification system was used to identify the pathogens and drug susceptibility testing .The data were analyzed by WHONET 5 .6 software .Results A total of 231 strains of pathogens were isolated from urine cultures ,including 51(22 .1% ) strains of gram‐positive bacteria ,170(73 .6% ) strains of gram‐negative bacteria ,and 10(4 .3% ) strains of fungi;Escherichia coli ,the coagulase‐negative Staphylococcus ,Klebsiella pneumoniae and E .faecalis were ranked the top four species of pathogens ,accounting for 48 .9% ,13 .0% ,9 .5% ,5 .2% ,respectively .MRCNS among CNS were 73 .6% .Staphylococcus had 100 .00% sensitivity to vancomycin and linezolid ;the antimicrobial resistance rate of E .fae‐calis to ampicillin was 8 .3% .E .faecalis had 100 .00% sensitivity to vancomycin and linezolid .the detectable rates of Escherichia co‐li ,Klebsiella pneumoniae of ESBLs were 52 .2% and 50 .0% ,and the strains had 100 .00% sensitivity to imipenem and meropenem . Conclusion Escherichia coli is a major pathogen in urine culture ,Bacterial resistance is serious .

ObjectiveTo explore the clinical significance of fms-like tyrosine kinase3(FLT3) expression in evaluation prognosis of acute myeloid leukemia(AML) prognosis.Methods50 patients with AML were selected.AML patients with normal karyotype were 20 cases,the abnormal karyotype were 30 cases.3ml bone marrow before themotherapy was aspirated respectively,and the FLT3 gene expression in leukemia cells was detected with polyenzyme chain react(PCR).ResultsThe FLT3 expression rate in AML patients with normal karyotype was 5.0％,and was 26.7％ in AML patients with abnormal karyotype,33.3％ in AML patients with refractory-relapse,and 4.5％ in AML patients with continue remission.The FLT3 expression rate was related with high leukemia cells percentage in bone marrow and high blood cells count in peripheral blood,and was not related with Franch America British(FAB) classification.The free-disease survival(FDS) and overall survived(OS) was shorter in FLT3 expression AML patients than that in no FLT3 expression AML patients.There was a statistical significance between the former and the latter( x2 =4.17,P ＜0.05 ).Conclusion FLT3 was a kind of worse factor in AML patients prognosis,and could guide clinical individual treatment in AML.

Objective To investigate the effective indicators for the prognosis assessment in pa?tients with multiple myeloma (MM) by 18F?FDG PET/CT imaging. Methods A total of 36 patients(22 males, 14 females;median age 63.5 years) with MM confirmed by clinical or pathology from July 2007 to November 2014 were retrospectively reviewed. The number of lesions detected by PET/CT, the number of lesions with SUVmax>2.5, the SUVmax and MTV of each lesion were calculated. The correlation analysis was performed between the number of lesions detected by PET or CT,the number of lesions with SUVmax>2.5, the SUVmax , MTV and serumβ2?microglobulin (β2?M) , respectively. The patients were divided into differ?ent groups according to the development of lesions and the survival situation during the follow?up ( 4-92 months) . Kaplan?Meier analysis and multivariate Cox model were used to analyze the prognostic significance of the number of lesions detected by PET or CT and the number of lesions with SUVmax>2.5, the SUVmax and MTV. Results Both the number of lesions with SUVmax>2. 5 and MTV showed positive correlations with blood β2?M (r=0.776, 0.954, both P<0.001), while the number of lesions detected by PET/CT and SUVmax were not correlated with β2?M ( r=0.053, 0.063, 0.398, all P>0.05) . The number of lesions with SUVmax>2.5 and MTV in the progressive group( n=14) were significantly higher than those in the regressive group(n=22):66.57±4.59 vs 31.95±4.75, t=4.95, P<0.001;(287.54±31.94) cm3 vs (72.17±14.35) cm3, t=6.93, P<0.001. The number of lesions with SUVmax>2.5 and MTV were significantly higher in the dead group(n=15) than those in the survival group(n=21):65.73±4.32 vs 30.90±4.87, t=5.10, P<0?001;(267.28±34.89) cm3 vs (76.39±15.67) cm3, t=5.49, P<0.001. The best cutoff values for predicting pro?gression?free survival and overall survival were both 42 for the number of lesions with SUVmax>2. 5, and those were 114.74 and 105.48 cm3 for MTV, respectively. The progression?free survival rate was worse in the patients with higher index than those with lower value (χ2=18.20, 29.74, both P<0.001) , and the same re?sult was also seen for the overall survival rate (χ2=19.07, 25.34, both P<0.001) . Conclusion The number of lesions with SUVmax>2.5 and MTV on 18 F?FDG PET/CT images could predict the progression?free survival and overall survival rates of patients with MM, which may provide accurate prognosis information.

Objective To investigate the correlation between admission hyperglycemia and clinical prognosis in elderly patients with community-acquired pneumonia.Methods Based on the electrical medical record (EMR),information of the patients with pneumonia was retrospectively collected and the necessary data were prepared and arranged.Based on clinical outcomes,demographic characteristics and clinical indicators were compared between groups in order to screen potential variables affecting clinical outcomes.Then,the random blood glucose level 8.6 mmol/L was considered to be the cutoff point for grouping patients,with clinical outcomes as outcome variables,random blood glucose levels as grouping variables,propensity score value analysis method was applied along with the ratio of 1:1 nearest neighbor matching method to generate paired samples in order to assess the impact of blood glucose level on mortality rate;With hospitalization time serving as time variable,survive analysis method was used to analyze COX proportional model.Results Five hundred and fifty-eight patients were enrolled in the analysis,197 patients of which were recorded dead during hospitalization (the dead group).After the correction of demographic characteristics,past medical history and blood test indexes,white blood cells,C reactive protein,random blood glucose at admission and triglyceride levels were regarded as potential factors for the deaths of patients (P<0.05).However,the propensity score analysis method suggested that there was no significant difference between the blood glucose levels and mortality.The survival analysis showed that the mortality in the high blood glucose group was higher than that in the normal blood glucose group (HR=1.68,95%CI(1.22,2.31)).Conclusion Admission hyperglycemia is an important indicator of adverse prognosis in elderly patients with pneumonia.It may be of significance for screening and identifying high-risk patients.

ObjectiveTo investigate the effect of macrophages (MCs) on the differentiation of mouse induced pluripotent stem cells (iPSCs) into hepatic progenitor cells (HPCs). MethodsA total of 24 C57BL/6N mice were used to obtain MCs by peritoneal irrigation, and the supernatant was collected to obtain the conditioned medium of MCs (MC-CDM). Activin A, bone morphogenetic protein 4, and fibroblast growth factor were used to induce the differentiation of mouse iPSCs into HPCs. The differentiation of HPCs were randomly divided into control group (normal medium) and experimental group (MC group; use of MC-CDM medium on day 5 of induction). Morphology, immunofluorescence assay, and Western blot were used to compare the morphology of HPCs and the expression of related proteins between the control group and the MC group. The t-test was used for comparison of continuous data between two groups. ResultsHPCs derived from iPSCs were established in vitro, and HPCs had the potential to differentiate into hepatocytes. Immunofluorescence assay showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-specific protein CK19 (0.901±0.072 vs 0.686±0.097, t=-3.093, P＜0.05). Western blot showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-related protein CK19 (1.922±0.103 vs 1.448±0.012, t =-7.881, P ＜005), as well as a significant increase in the protein expression of the autophagy-related protein LC3 (1.392±0.042 vs 1.101±0048, t =-5.978, P＜005). ConclusionMCs can promote the differentiation of mouse iPSCs into HPCs, possibly by increasing the autophagy level of HPCs.

Objective To investigate the value of IL-27,IL-29 and miRNA-497 in HER-2 positive radioactive particles combined targeted therapy. Methods Sixty-one elderly inpatients with HER-2 positive gastric cancer ascites in the Gaoming District People's Hospital, Wuchuan Municipal People's Hospital and Zhejiang Provincial Tumor Hospital were divided into the two groups. The treatment group(n= 31) was treated by the radioactive particles+ targeted drug Trastuzumab(initial dose 8 mg/kg, maintenance dose 66 mg/kg, once per 3 weeks) for 6 weeks; the control group(30 cases) was treated by targeted drug Trastuzumab (initial dose 8 mg/kg,maintenance dose 6 mg/kg, once per 3 weeks) for 6 weeks. The levels of ascites IL-27,IL-29 and miRNA-497 in the radioactive partcles combined targeted therapy group and control group were detected. Results The levels of ascites IL-27, IL-29 and miRNA-497 after treatment in the treatment group were significantly decreased compared with those before treatment and in the control group,the effective remission rate in the treatment group was 74. 19%, which was higher than 36.67% in the control group, the average median survival time in the treatment group was 155 d, which washigher than 72 d in the control group, the difference was statistically significant. Conclusion IL-27 ,IL-29 and miRNA-497 has the significance of curative effect evaluation and prognosis judgment in the radioactive particles combined targeted therapy for the patients with HER 2 positive gastric cance.

Objective To evaluate the long-term outcome of fully covered self-expandable metal stents(FCSEMS)for the treatment of benign biliary strictures(BBS). Methods Between June 2008 and September 2013, 68 patients with BBS receiving endoscopic retrograde cholangiopancreatography and FCSEMS placement were retrospectively enrolled. Data of endoscopic treatment, stricture resolution and recurrence were collected, and related risk factors were analyzed. Results FCSEMSs were successfully placed in all patients and removed in 93.4%(57/61). The median stent duration was 9.0(range 0.2-37.1)months. Stricture resolution occurred in 74.2%(46/62)patients. During median follow-up of 54.0 (range 2.5-96.0)months,stricture recurrences were seen in 16.7%(6/36)patients. Multivariate analysis revealed that distance between stricture and hepatic bifurcation of less than 1.5 cm(P=0.034,OR=6.395, 95%CI:1.153-35.464), and stent migration(P= 0.024, OR= 0.153, 95%CI:0.030-0.782)were significant risk factors for stricture resolution. Meanwhile, the stricture length longer than 1.0 cm(P=0.028, HR = 6.766, 95% CI:1.233-37.122)was a significant risk factor for stricture recurrence. Conclusion Endoscopic treatment combined with FCSEMS can achieve excellent efficacy in resolving BBS with low recurrence rate. However, location and length of BBS, as well as stent migration may impair its effectiveness.

As a new type of medical service mode, day surgery significantly alleviates the contradiction between supply and demand of medical resources. Since 2009, day surgery center in West China Hospital of Sichuan University has gradually carried out endoscopic gastrointestinal(GI) polypectomy, continuously explored and practiced the centralized management mode of GI polyps. The integrated management process included pre-hospital management, clinical pathway during hospitalization, and extended service after discharge. The overall-management database for day surgery patients was established, guiding the clinical practice and standardizing clinical behavior based on various clinical studies, which help day surgery managers make clinical decisions, and achieve the management objectives of fine management, quality and safety normalization for day surgery.