Objective To culture human sapovirus(HuSaV) GⅠ.1 in vitro and prepare polyclonal antibody against the capsid protein VP1.Methods HuSaV GⅠ.1 positive stool specimens preserved in diarrhea department of National Institute for Viral Disease Control and Prevention were inoculated with HuTu-80 cells supplemented with different bile acid salts[glycine chenodeoxycholic acid(GCDCA) and glycine cholic acid(GCA)],and the infection,proliferation and passage of the virus were determined by PCR and RT-qPCR.The VP1 gene was amplified by PCR and cloned into prokaryotic expression vector pGEX-6P-1.The constructed recombinant expression plasmid pGEX-6P-1-VP1 was transformed into E.coli BL21(DE3) and induced by IPTG.Two female New Zealand white rabbits were immunized with the purified recombinant VP1 protein for 4 times.The blood samples were collected 18,28,38 and 48 d after immunization,and the serum titers were detected by ELISA.Results HuTu-80 cells were effectively infected by HuSaV GⅠ.1 in the presence of bile acid salt GCA,and the proliferated virus were stably and continuously transmitted for three generations in HuTu-80 cells.The expressed recom-binant GST-VP1 protein showed a relative molecular mass of about 86 000,and about 60 000 after purification(GST tag excision).The titer of polyclonal antibody against HuSaV VP1 protein was over 1:12 800.Conclusion HuSaV was successfully isolated and cultured in vitro using HuTu-80 cells supplemented with bile acid salt,and polyclonal antibody with high titer against HuSaV VP1 protein was prepared,which laid a foundation of in-depth research of HuSaV identification,infection and pathogenesis.

Objective To investigate the curative efficacy of safflower yellow sodium chloride in the treat-ment of acute myocardial infarction (AMI)and its effects on cardiac function and oxidative stress.Methods 110 cases of AMI meeting the inclusion criteria were randomly divided into two groups equally.The control group was given conventional drug treatment and percutaneous coronary intervention (PCI),while the observation group was given con-ventional drug treatment and PCI in combination with safflower yellow sodium chloride.Then,the curative efficacy, indices of cardiac function and oxidative stress were observed and compared.Results The observation group had a total effective rate of 94.5%,which was significantly higher than 81.8% in the control group (χ2 =4.94,P =0.026).After the treatment,SV (t =3.23,P =0.002),CO(t =3.82,P =0.000)and LVEF (t =3.14,P =0.002) in the observation group were significantly higher than those in the control group (all P <0.05 ).As to oxidative stress,in comparison with the control group,SOD (t =8.42,P =0.000)was significantly higher and MDA (t =3.99, P =0.000)was significantly lower in the observation group (all P <0.05 ).Conclusion Adjuvant treatment of safflower yellow sodium chloride for AMI was effective,which could significantly increase curative efficacy,improve cardiac function and alleviate oxidative stress.

Objective To observe the serum estradiol level in newborns at different gestational age and the infants with retinopathy of prematurity (ROP), and explore the relationship between these changes and ROP. Methods A total of 178 newborns hospitalized in the Children's Hospital of Fudan University were enrolled in the study. They were divided into three groups based on the gestational age, ＜ 32 weeks group, 32-37 weeks group and term group. The three groups were further divided into non-oxygen and oxygen subgroups. Infants with birth weight less than 2000 g were evaluated for ROP and those diagnosed as ROP were under ROP group (n=6). Serum estradiol levels were measured every week from birth to discharge with radioimmunoassay. Results Serum estradiol levels of newborns quickly declined after birth. In the non-oxygen subgroups, serum estradiol levels in 32 ～ 37 weeks group were remarkably higher than those of term group in the first week [(3506±1376) pmol/L vs (1717±179) pmol/L,P＜0. 05] and the third week [(1431±92) pmol/L vs (996±102) pmol/L, P＜0. 05]; Serum estradiol levels in ＜32 weeks group was only remarkably higher than those of term group in the first week [(3173± 1369) pmol/L vs (1717± 179) pmol/L,P＜0. 05]; Serum estradiol level in the ＜32 week group was (560±355) pmol/L, lower than that in the 32-37 weeks group [( 1124 ± 128) pmol/L] (P＜0. 05). There was no difference of serum estradiol levels in the oxygen and non-oxygen subgroups of the same group. There was no significant difference of serum estradiol levels between the ROP group and ＜32 weeks group. Serum estradiol levels in ROP group was lower than that of premature group without ROP at 29 and 32-36 weeks of corrected gestational age, but no statistical difference was shown. Conclusions Very low serum estradiol level in premature infants may be related to the development of ROP, but further clinical studies are required.

Objective To explore the factors related to the outcome of idiopathic facial palsy. Methods 308 patients with idiopathic fa-cial palsy were included. The data of clinic and follow-up were collected from January, 2010 to December, 2014. The data were analyzed by Logistic regression analysis. Results 210 cases (68.2%) were cured (good group) and 98 cases (31.8%) appeared sequelae of different de-grees (poor group). The age, onset of disease, type of disease, damaged section, impaired glucose control, high neutrophil-to-lymphocyte ra-tio (NLR), high blood glucose, high blood triglyceride, time and methods of invention were significant factors related to the outcome (P<0.05). Conclusion Old, servious facial nerve injury, Hunt's palsy, high damaged section, poor glucose control in the patients with diabetes, high NLR, high blood triglyceride, delay and simple invention are independent risk factors for the poor outcome of idiopathic facial palsy.

This research analyzed the current situation of English learning anxiety of TCM majors by means of conducting a questionnaire based on the English learning anxiety scale (ELAS).The questionnaire results showed that communication apprehension was most prominent in terms of extent and amount while test anxiety also accounted for an important part of anxiety.The research further analyzed the influences of English learning anxiety and proposed some measures to ease students' anxiety and to enhance learning efficiency from teachers' perspectives.

Objective To confirm the structure validity of questionnaire through a probe into latent factorial structure of the rear service personnel′s job satisfaction from polyclinics hospitals .Methods The questionnaire′s construct validity was estimated by ex‐ploratory factor analysis (EFA) and confirmatory factor analysis(CFA) .Results Exploratory factor analysis made the question‐naire including six factors which belonged to job satisfaction and occupation loyalty .Every factor had reasonable explanation and six factors remained 70 .306% information .Exploratory factor analysis of job satisfaction showed five factors ,explaining 70 .514% in‐formation .The confirmatory factor analysis model 1 and model 2 were conducted by LISREL8 .70 software .The results showed that X2 freedom degree was less than 5 .In addition to adjust the goodness of fit index was less than 0 .9 ,all other fit indices were greater than 0 .9 .At the same time ,SRMR and RMSEA were less than 0 .08 .Conclusion The results of exploratory factor analysis(EFA) and confirmatory factor analysis(CFA) both reveal that the questionnaire′s construct validity was good .

Objective:To transfect the tongue squamous cell carcinoma Tca8113 cells with NICD, and to investigate the effects of Notch 1 on Tca8113 cells and the expression of epidermal growth factor receptor (EGFR) in tongue squamous cell carcinoma.Methods: The Notch plasmid pRAMIC-IRES2-EGFP encoding exogenous and control plasmid pIRES2-EGFP were transiently transfected into the Tca8113 cells by LipofectamineTM 2000. In the experiment, target plasmid pRAMIC-IRES2-EGFP transfection group, control plasmid pIRES2-EGFP transfection group and non-transfection group were set up.RT-PCR, Western blotting and immunocytochemical method were used to observe the mRNA and protein expression levels of Notch 1 and EGFR.The proliferation activities of Tca8113 cells were detected by MTT assay,and the survival rates were calculated.Results:Compared with non-transfection group and control plasmid pIRES2-EGFP transfection group, the mRNA and protein expression levels of Notch 1 in Tca8113 cells in pRAMIC-IRES2-EGFP group was significantly increased(P<0.05), the mRNA and protein expression levels of EGFR in Tca8113 cells were significantly decreased (P<0.05),and the proliferation rate of Tca8113 cells was significantly decreased(P<0.05) after transfection.There were no significant differences in the expression levels of Notch 1 and EGFR mRNA and protein and the proliferation rates in Tca8113 cells between non-transfection group and pRAMIC-IRES2-EGFP group (P>0.05).Conclusion:The effect of Notch 1 on the proliferation of Tca8113 cells may be related to down-ragulatoin of expression level of EGFR.

Objective To analyze the application value of high risk human papillomavirus (HR HPV) DNA and thinprep cytolo-gy test(TCT ) detection in screening cervical cancer and to evaluate the clinical significance of their combined detection .Methods A total of 402 cases conducting cervical cancer screening in our hospital from March to May 2016 were selected and performed the HR HPV DNA and TCT detection .Those of suspected cervical lesions were performed the histopathological examination .Then the effects of HR HPV DNA and TCT combined detection were compared according to the pathological results .Results The detection positive rate of female HR HPV DNA was 27 .1% (109/402);abnormal and benign inflammation reaction were detected out in 46 cases of female TCT ,the positive rate was 11 .4% (46/402);in 123 cases of suspected cervical malignant lesion ,32 .5% females(40/123) appeared CIN Ⅰgrade or more lesions by histopathological examination ;the sensitivity of HR HPV DNA and TCT combined detection was higher than that of single detection ,the difference was statistically significant (P<0 .05);the specificity of HR HPV DNA and TCT combined detection was higher than that of HR HPV DNA single detection ,the difference was statistically signifi-cant(P<0 .05) ,but the difference was not statistically significant compared with TCT single detection (P>0 .05) .Conclusion The detection of HR HPV DNA and TCT is a good method for screening of cervical cancer ,the combined detection of HR HPV DNA and TCT can improve the detection rate of cervical precancerous lesion .

Objective To systematicly evaluate the effectiveness of terminal care for the elderly patients with tumor in China. Methods Chinese databases including China National Knowledge Internet (CNKI),VIP database,Wanfang Data,China Biology Medicine (CBM) were searched to collect the randomized control trials (RCTs) from inception to November 2014. According to the inclusion and exclusion criteria, the literatures were screened, datas were extracted, and methodological qualities were assessed, then a Meta-analysis was performed by using RevMan software, version 5.3. Results 8 RCTs involved 557 patients were included. The results of this Meta-analysis showed that the elderly tumor patients′quality of life in terminal care patients were superior to that in routine care patients with statistically significant difference [ standardized mean difference was 4.78, 95%confidence interval (95%CI) 2.06-7.50, P<0.01). The satisfaction of patients and their family members in terminal care patients were superior to that in routine care patients and there were statistically significant difference (relative risk was 1.55, 95%CI 1.20-2.00, P<0.01). Conclusions For elderly tumor patients, terminal care can improve their quality of life, and improve the satisfaction of patients and their families.