The design of DNA-based alphavirus vectors significantly improves the utility of these replicon vectors. The DNA-based replicon vectors can be used in expressing foreign genes and preparing RVP in virto efficiently, also in developing replicon vaccines and gene therapy vectors in vivo. The approach involved the conversion a RNA-based replicon vector into a layered DNA-based replicon vector by the RNA polymerase Ⅱ promoter and transcription termination/polyadenylation signal transcribed replicon RNA from DNA. When DNA-based alphavirus vector tranfected into cells, the first layer includes a eukaryotic RNA polymerase Ⅱ expression cassette that initiates transcription of RNA in nucleus. Following transport of this RNA from the nucleus to the cytoplasm, the second layer, autocatalytic amplification of the RNA vector corresponds to virus RNA replication cycle and results in high level expression of foreign gene. DNA and RNA-based bifunctional replicon expression vector pSCTA and helper vector pSHCTA were successfully constructed by replacing the SP6 promoter used in the original system pSFV1 and pSFV-helper2 derived from Semliki Forest virus (SFV) with CMV promoter and T7 promoter, and inserting BGH transcription termination and polyadenylation signal downstream 3′-untranslated region (UTR). In order to obtain DNA-based highly efficient replicon vectors, they were further modified to construct additional three DNA-based SFV replicon expression vectors and corresponding helper vectors. To investigate the efficiency of foreign gene expression level by the four different DNA-based SFV expression vectors and recombinant virus particle (RVP) prepared by cotranfecting with corresponding helper vectors, improved DNA-based replicon vectors pSCAR and pSHCAR derived from SFV were developed. high level protein could be generated using the new vector system by transfecting DNA into BHK21 cells and High titer of RVP produced by cotranfecting with helper vector. Antigen genes were also expressed in cells by the replicon expression vector. Additionally, reporter gene expression was observed in mice muscle following injection with SFV DNA vector. Anti-?-Gal antibody response and cell-mediated immune response were induced after intramuscular inoculation of the ?-Gal-encoding SFV replicon DNA. The results suggested that highly efficient DNA-based replicon vectors pSCAR and pSHCAR were constructed by modifying the SFV vectors. The improved DNA-based replicon vectors enhance the utility of them, and can be developed as potentially replicon vaccines and gene therapy vectors.

Objective To explore the association between maternal age and perinatal outcomes.Methods Totally,3 151 women with advanced maternal age and 6 098 women younger than 35 years old who delivered in Beijing Obstetrics and Gynecology Hospital in 2016 were recruited.Their clinic characteristics and perinatal outcomes were collected to divide into 3 groups based on delivery age,Group 1 (aged 35-39 years,2 683 cases),Group 2 (aged ≥40 years,366 cases) and the control group (aged＜35 years,6 098 cases).The association between maternal age and adverse perinatal outcomes were analyzed,including hypertensive disorder complicating pregnancy,gestational diabetes mellitus (GDM),preterm birth and postpartum hemorrhage.Results The rate of cesarean section history (27.39％,33.61％,5.53％) or previous myomectomy history (2.80％,5.46％,0.72％) were compared between the advanced maternal age groups and the control group,and the differences were statistically significant (P＜0.05).The percentage of prepregnancy overweight and obesity (29.67％,27.05％,18.47％),complicated with myoma (14.83％,19.95％,5.64％) were compared among the three groups,and the differences were statistically significant (P＜ 0.05).The percentage of pregnancy through assisted reproductive technology (9.84％,15.03％,3.12％) also had statistically significant differences (P＜0.05).The incidence of fetal chromosomal abnormalities (1.23％,3.01％,0.36％) and fetal malformations (1.94％,4.37％,0.48％) increased with the maternal age,with statistically significant differences (P＜0.01).The mobidity of hypertensive disorders (9.84％,13.11％,9.23％),pregestational diabetes mellitus (1.83％,2.19％,0.72％),gestational diabetes mellitus (22.70％,28.42％,14.87％),premature rupture of membranes (25.57％,19.40％,31.42％),placenta previa (2.05％,2.46％,0.92％),preterm birth(8.35％,11.20％,5.51％),postpartum hemorrhage (25.11％,18.31％,20.27％)and forceps delivery (5.42％,2.33％,5.71％) were compared,and the differences were statistically significant (P＜0.05).The cesarean section rate in primipara (45.42％,75.74％,21.33％) and multipara (51.46％,61.54％,30.95％) had statistically significant difference (P＜0.05).The proportion of macrosomia (10.80％,8.85％,7.96％) and neonates transferred into neonatal ICU (9.63％,11.48％,5.21％) in term neonates had statistically significant difference (P＜0.05).Conclusions Women with advanced maternal age increase after new family planning policy put into effect,so do the risk of adverse perinatal outcomes.Attention and interventions should be made to cope with the occurrence of adverse perinatal outcomes.

Objective To study the coexpression of protein folding modulators,including chaperones(DnaK,DnaJ,and GroEL-GroES) and disulfide isomerase Dsb molecules(DsbA and DsbC) on the bioactivity of eukaryotic expressed urokinase(UK).Methods The gene sequences of DnaK,DnaJ,GroEL-GroES,DsbA and DsbC were isolated from the chromosome of E.coli by PCR.The obtained fragments mentioned above were cloned into the plasmid pBAD-1 under the control of araB promoter and then the plasmids were transformed into E.coli BW25113 and HB101 together with a compatible vector pQE-UK.The amount of expressed UK was measured by Bradford method and its activity was analyzed by fibrin plate method for its fibrinolysis.Results The activity of expressed UK was increased by co-expressing with DnaK,GroEL-GroES,DsbA,and DsbC in BW25113.Among them,the highest activity was achieved by co-expressing with GroEL-GroES.Conclusion Co-expressing of chaperones or disulfide isomerase can improve the bioactivity of UK expressed in E.coli.

Objective To evaluate the application of scenario-based training and medical simulator in the orthopedics education. Methods A total of 60 students from Shanghai Jiao Tong University School of Medicine who finished their orthopedics internship from January 2016 to July 2016 were involved. They were randomly divided into the study group and control group with 30 students each. The study group re-ceived 2 classes of scenario-based training and medical simulator assisted education during their internship in the orthopedics department while the control group received 2 classes of traditional lessons instead. Sur-veys were conducted after the internship and the scores of internship were also recorded. Result The overall satisfaction was higher in the study group than the control group [(8.6±0.6) vs. (8.1±0.5), P=0.001]. On the part of learning interest, clinical thinking, clinical practice and group working, the study group also received better evaluation (P<0.05). The study group achieved better scores in the final examination than the control group [(84.4±2.6) vs. (82.5±3.4), P=0.018]. Conclusion The combination of scenario-based training and medical simulator can improve the ability of medical students in the orthopedics education, and receive higher satisfaction.

Objective To explore the relationship between periodontal disease and preeclampsia and the effects of periodontal treatment on preeclampsia.Methods China National Knowledge Infrastructure,Chinese Biomedical Literature Database,WANGFANG DATA,China Dissertation Full-Text Database,China Proceedings of Conference Full-Text Database,Cochrane Library,PubMed,EMbase,Elsevier,Springer,and Science Direct OnSite were extracted from inception till September 30,2014.The case-control,cohort and randomized controlled trials about the association of matemal periodontal disease and preeclampsia were searched according to the inclusion and exclusion criteria.RevMan5.1 and Stata12.0 were used to test the heterogeneity of the results among the different studies and amalgamate the effect size using fixed or random effect models.Results Twenty studies (15 case-control and 5 cohort) involving 8 775 women assessed the association between periodontal disease and preeclampsia.A positive association was found (OR=2.48,95％CI:1.76-3.48,P ＜ 0.01).Meta-analysis of the case-control studies showed more than twice in the odds of preeclampsia with the presence of periodontal disease (OR=2.75,95％CI:1.93-3.92,P ＜ 0.01).Meta-analysis of cohort studies did not reveal any significant differences (OR=1.84,95％CI:0.91-3.74,P ＞ 0.05).Four randomized controlled trials with 3 712 women evaluated the effect of periodontal treatment on preeclampsia,and meta-analysis showed no relative risk reduction in preeclampsia with periodontal treatment (RR=1.04,95％CI:0.84-1.30,P ＞ 0.05).Conclusions Periodontal disease appears to be a possible risk factor for preeclampsia,but treatment during pregnancy does not prevent preeclampsia.High-quality prospective studies are needed to confirm the relationship between periodontal disease and preeclampsia.

Objective To explore the value of magnetic resonance cholangiopancreatography (MRCP) and MRI in the diagnosis and differential diagnosis of periampullary carcinoma. Methods Plain and dynamic multi-phase enhanced MRI data of 54 patients with periampullary carcinoma proved by pathology were retrospectively analysed. χ~2 test and two independent samples t test were used to examine the relative results. The results of MRI were compared with that of pathology. Results Of 32 cases with carcinoma of head of pancreas, 7 cases (21.9%) exhibited "four-duct sign". 16 cases were carcinomas of the lower part of common bile ducts, 9 cases (56.3%) showed "three-duct sign". The difference between carcinomas of head of pancreas and carcinomas of the lower part of common bile ducts was significant (P<0.05). Additionally, the ratio of the largest area of masses and the diameter of com-mon bile duct in pancreatic cancer tumors was larger than that in common bile duct cancer (P<0.05) for whether the intraoperative or MRI measurement. Conclusion MRI and MRCP are very helpful in diagnosis of periampullary carcinoma.

Objective To investigate genes involved in the mechanisms underlying the progression of severe preeclampsia.Methods We conducted a muhiregional gene expression analysis using peripheral leucocytes from patients with preeclampsia and normal controls.Total RNA was extracted from peripheral blood of six severe preeclampsia and five normotensive pregnancies.We performed genome-wide expression profiling using Affymetrix HG_U133 plus 2.0 chips to screen out differentially expressed genes of 2 fold or more and q_value ＜ 5.4%.Using Gene Ontology we identified the function of differentially expressed genes after cluster analysis.Results Among the 47 000 genes that were screened in the microarray,140 genes were found to be differentially expressed between normal and preeclamptic pregnancies. Eighty six up-regulated candidate genes were mainly involved in cysteine metabolism urea cycle and metabolism of amiogroups,proteasome,TGF-beta signaling pathway, and the ratio of calponin2 (CNN2), matrix metallopeptidase 8 (MMP8),V-set and immunoglobulin domain containing 4 (VSIG4),proteasome 26S subunit ATPase 5 (PSMC5) was evidently increased in preeclampsia patients.Among 54 down-regulatedcandidates,natural killer cell mediated cytotoxicity,antigen processing and presentation,metabolism of xenobiotics by cytochrome P450 were the main pathways.KIR3DL2,AKR1C3,CHURC1 and SLC25A13 were obviously decreased in preeclampsia patients. Conclusions The gene expression of peripheral leucocytes in preeclampsia patients is significantly different from that of uncomplicated pregnancies.CNN2,MMP8,VSIG4,PSMC5,KIR3DL2,AKR1C3,CHURC1 and SLC25A13 may be involved in the molecular mechanisms underlying the progression of severe preeclampsia.

Objective To explore the immunogenicity of recombinant chimeric 6Aβ15-T including the Aβ1-15 epitope and a T-helper epitope formulated with different adjuvants and to evaluate its feasibility as a candidate vaccine for Alzheimer disease (AD).Methods The recombinant chimeric antigen 6Aβ15-T formulated with Al adjuvant, Freund′s adjuvant or MF59 adjuvant was administered to two strains of mice .The 6Aβ15-T-immunized group without adjuvants ( Mock) and non-immunized group (Control) were included in this study as control groups .The specific antibody and cellular immune response of the chimeric antigen were evaluated .Results In BALB/c strain mice, three types of adjuvants could substan-tially boost the immunogenicity of chimeric antigen 6Aβ15-T and produce a high level of specific-Aβ(β-amyloid) antibod-ies.In C57BL/6 strain mice, the existence of adjuvants enhanced the immune response of 6Aβ15-T antigen, but the mice in Mock group also produced a strong antibody response .In two strains of mice, prevalence of anti-AβIgG1, which was an indicator of Th2 polarization, was observed in the 6Aβ15-T-immunized mice.Additionally, the Al adjuvant induced a high-er level of IgG1 antibody titers, and the ratio of IgG1/IgG2a was the largest.As expected, the 6Aβ15-T antigen formulated with or without adjuvants induced PADRE-specific, but not Aβ42-specific T cellular immune response .Conclusion The 6Aβ15-T antigens formulated with different types of adjuvants could induce strong Th 2-polarized Aβ42-specific antibody re-sponses without activating self-reactive Aβ42-specific T cells in two strains of mice .The results suggested that the recombi-nant chimeric antigen 6Aβ15-T is a good candidate vaccine for AD .

AIM:To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism.METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer ( GC) tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS:Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples ( P<0.05 ) .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05).Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased (P<0.05), the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05).CONCLUSION:The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.

Background and purpose:It has beenreported that miR-1284 is associated with gastric cancer lymph node metastasis in the research of microRNA microarray in human gastric cancer tissues. But the specific role of miR-1284 in gastric cancer has not been reported. The aim of this study was to investigate the effect of miR-1284 over-expression on the gene expression profiling and invasion/metastasis of human gastric cancer SGC-7901 cells. Methods:Gastric cancer SGC-7901 cells of LV-miR-1284 group were transfected with lentiviral vectors of miR-1284, cells of LV-NC-GFP group were transfected with lentiviral vectors without miR-1284, and cells of control group were not transfected with lentiviral vectors. The expression of miR-1284 was detected by the real-time fluorescent quantitative PCR. Differential expression genes were detected by the microRNA chip. Target genes of miR-1284 were predicted by the bioinformatics. Invasive ability was detected by the Transwell invasion assay. Metastasis ability was detected by subcutane-ously transplanted tumor model of nude mice.Results:Compared with LV-NC-GFP and control groups, the expressions of miR-1284 and 20 genes were up-regulated, and the expression of 17 genes was down-regulated in LV-miR-1284 group. One hundred and thirty-eight target genes of miR-1284 were predicted by the bioinformatics website. Compared with invasive cell number of LV-NC-GFP group (168.67±4.55) and control group (170.33±3.08), the ability of invasion ofcells was weakened in LV-miR-1284 group (70.00±2.37). Compared with the liver metastasis rate of LV-NC-GFP group (85.71%) and control group (85.71%), the ability of metastasis of cells was weakened in LV-miR-1284 group (14.29%). Conclusion:The ability of invasion and metastasis of SGC-7901 cells is suppressed by over-expression of miR-1284. The mechanism may be related to regulating the expression ofSUMO1 andJUNgenes.