Dengue (DEN) virus is a member of the family flaviviridae which consists of a group of enveloped viruses with a single-stranded positive-sense RNA genome. DEN virus infection is initiated by the binding of envelope protein E to cellular receptor,followed by replication in cytoplasm which associated with a range of induced membrane structures. Further characterization of the DEN replication complex (RC) showed the involvement of NS1, NS2A, NS3, NS4A, and NS5 proteins as the viral replicase. The DEN virion maturation is associated with the cleavage of precursor of protein C by NS2B/NS3 protease and the glucosylase-mediated N-linked oligosaccharide trimming of prM, E protein. This review summarizes recent advance in research on DEN virus life cycle, emphasizing cellular receptor, RNA replication and virion assembly.

ObjectiveTo study the level of temperament and self-esteem of children aged 8～12 in Beijing city,as well as their interaction and influenced factor.Methods203 school children aged 8～12 were assessed with Carey's temperament questionnaire(8～12:MCTQ) and The Self-Esteem Scale(SES).ResultsTemperament and self-esteem of school children aged 8～12 were associated with the parental behavior,especially with work style of their fathers.The rhythmicity was found to be negative correlated to the self-esteem scores,and the intensity of reaction was found to be positive correlated to the self-esteem scores.ConclusionParents' behavior and family environment were important to the temperament and self-esteem of school children.

Allergic rhinitis (AR) is a noninfectious inflammatory response in nasal mucosa caused by allergens, which is contacted by a specific individual. The immune imbalance of Th1/Th2 plays an important role in the pathogenesis of AR. In?terleukin (IL)-33, the novel cytokine of IL-1 family, is an important regulatory factor of allergic diseases, autoimmune diseas?es and various inflammatory diseases. IL-33 is a kind of alarm, which is mainly secreted and released by damaged tissues and cells, especially impaired epithelial cells and endothelial cells. IL-33 binding to its receptor ST2L can activate a variety of immune cells to produce Th2 cytokines, precipitating and maintaining Th2 polarization, increasing AR immune inflamma?tion, which is the new target of AR in research and treatment. In this article, we have done a brief overview for the biological functions of IL-33 and its receptor ST2L and the research progress in the AR.

This paper investigated contamination situation of Listeria monocytogenes(Lm). To compare dif- ferent selectivity enrichment broth for detectable effect of Lm and compare detectable effect in different samples by using different methods, furthermore, choose the best enrichment broth for specific food. One hundred and thirty five random samples from raw meat, aquatic product, fruit and vegetable, quick-frozen food in Baoding. Applied LB enrichment broth, EB enrichment broth, new modification FDA enrichment broth and Fraser enrichment broth before separated by PALCAM selective agar, then identified by interna- tional standard method after PCR. Results: Four methods showed that there were 23 Lm positive, detected 5 Lm by LB method, 6 Lm by Fraser method, 5 Lm by EB method and 7 Lm by new modification FDA method. The total detectable rate of four methods had no large specificity, but to specific kind of food was different.

Parkinson's disease(PD) is a common neurodegenerative disorder with no effective protective treatment,characterized by a massive degeneration of dopaminergic neurons in the substantia nigra(SNpc) and the subsequent loss of their projecting nerve fibers in the striatum.The major neurochemical manifestation of this disorder is the loss of the neurotransmitter dopamine(DA) in the striatum as a result of the progressive degeneration of the dopaminergic neurons in the substantia nigra.There have been significant progresses in recent years reporting on the use of mesenchymal stem cells(MSCs)in gene therapy,with specific application towards PD.MSCs,a kind of multipotent adult progenitor cells,are considered as a useful vehicle for cell and gene therapy because of their multiple differentiation potentiality and self-transplantation.The present study was focused on treating rat model of PD using human tyrosine hydroxylase gene(hTH),human aromatic L-amino acid decarboxylase gene(hAADC) and human GT Pcyclohydrolase I gene(hGCH-I) engineered MSCs,in order to provide a better understanding about the application of these cells in the therapeutic benifit of PD.The gene of hTH,hAADC and hGCH-I were introduced via recombinant adeno-associated virus(rAAV) infection into the MSCs in vitro.The genetically modified MSCs expressing hTH,hAADC and hGCH-I were transplanted into the striatum of PD rat models.The behavior,the nigra-striatal level of DA and its metabolite were detected.The results of present study were shown as follows:hTH,hAADC,hGCH-I and LacZ gene were transfected into MSCs with adeno-associated virus vectors.The HEK293 packaging cells(ATCC) were transfected with the plasmids of pAAV-hTH,pAAV-hAADC,pAAV-hGCH-I,pAAV-LacZ,pAAV-RC,pHelper by using calcium phosphate precipitation.Titer was detected using HT1080 cells.Viral particles were collected and used to infect MSCs.The purified modified MSCs expressing the three kinds of genes were selected separately and were grafted in the striatum of the PD model rats in the lesion side.The MSCs genetically modified suvived well 12 weeks after transplantation.The improvements of the behavior were observed every week after transplantation.Compared with the control group,the rounds of asymmetric rotation after apomorphine administration decreased in the groups double or triple genes engineered MSCs grafted(p

Objective To investigate the effect of dexmedetomidine postconditioning on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats.Methods Fifty male Spragne-Dawley rats,aged 7-8 weeks,weighing 200-250 g,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C),LPS group and postconditioning with 3 different doses of dexmedetomidine groups (LD,MD and HD groups).ALI was induced with LPS 8 mg/kg injected via the caudal vein in LPS,LD,MD and HD groups.Dexmedetomidine 5,10 and 15 μg/kg were injected intraperitoneally in LD,MD and HD groups,respectively,at 1 h after LPS injection.The equal volume of normal saline was injected intraperitoneally in C and L groups.Blood samples were taken from the left ventricle at 6 h after dexmedetomidine administration,then the animals were sacrificed and broncheoalveolar lavage fluid (BALF) was collected.The concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in plasma and BALF were detected by ELISA.The lungs were removed for microscopic examination of pathological changes and for determination of wet/dry lung weight (W/D) ratio and expression of Toll-like receptor 4 (TLR4) mRNA (by RT-PCR) in lung tissues.Results Compared with group C,W/D ratio,pathological scores,and the concentrations of IL-6 and TNF-α in plasma and BALF were significantly increased,and the expression of TLR4 mRNA was up-regulated in LPS,LD,MD and HD groups.Compared with LPS and LD groups,W/D ratio,pathological scores,and the concentrations of IL-6 and TNF-α in plasma and BALF were significantly decreased,and the expression of TLR4 mRNA was down-regulated in MD and HD groups.There was no significant difference in the parameters mentioned above between LPS and LD groups,and between MD and HD groups.Conclusion Dexmedetomidine postconditioning can alleviate ALI induced by LPS in rats,and up-regulated TLR4 mRNA expression and reduced inflammatory responses may be involved in the mechanism.

Objective To investigate the preventive effects of aspirin on liver metastases of colorectal cancer in mice and study the mechanisms.Methods Twenty BALB/c mice were divided into the control group and the experimental group according to the random number table with 10 mice in each group.Mice in the control group were fed with saline each day at a concentration of 0.2 mL/d for 60 days,while mice in the aspirin group were fed with aspirin each day at a concentration of 30 μg/(g · d) for 60 days.Then C26 colon cancer cells were injected into the spleen and then the spleen was cut to establish mice model of colon cancer liver metastasis.The C26 colon cancer cells were divided into 2 groups.C26 colon cancer cells in the control group remained untreated,and C26 colon cancer cells in the experimental group were treated with aspirin at a concentration of 10 mmol/L for 24 hours.The scratches and transwell assays were conducted to observe the effects of aspirin on the invasion and metastasis of C26 colon cancer cells.The expressions of epithelial-mesenchymal transition (EMT)-related genes were detected using RT-PCR and Western blot.All data were analyzed using the Student t test.The survival curve was drawn by Kaplan-Meier method,and the survival analysis was done by Log-rank test.Results The numbers and weights of hepatic metastatic tumors were 4.8 ± 1.9 and (504 ± 107) mg in the control group and 2.6 ± 1.6 and (362 ± 67) mg in the experimental group,with significant difference between the 2 groups (t =2.840,3.584,P ＜ 0.05).The 1-month survival rate was 80％ in the experimental group,which was significantly higher than 40％ of the control group (x2=4.418,P ＜ 0.05).The results of pathological examination showed that tumor cell heteromorphism was reduced by aspirin.The results of scratches experiment showed an obvious migration of C26 colon cancer cells in the control group at 24 hours later,while no C26 colon cancer cells migrated in the experimental group.The numbers of C26 colon cancer cells penetrated the Watrige were 253 ± 21 in the control group and 148 ± 13 in the experimental group,with significant difference between the 2 groups (t =5.101,P ＜0.05).The relative mRNA expression of the E-cadherin and the Vimentin were 0.002 ±0.001 and 1.005 ±0.286 in the control group and 0.005 ± 0.001 and 0.270 ± 0.168 in the experimental group,with significant difference between the 2 groups (t =-4.606,4.942,P ＜ 0.05).The relative protein expressions of the E-cadherin and the Vimentin were 0.473 ±0.179 and 0.787 ± 0.118 in the control group and 1.585 ± 0.410 and 0.280 ± 0.133 in the experimental group,with significant difference between the 2 groups (t =-5.542,6.355,P ＜ 0.05).Conclusion Aspirin inhibits liver metastasis of colon cancer and promote the survival ratio of mice.Aspirin can up-regulate the expression of E-cadherin and down-regulate the expression of Vimentin,which inhibits EMT and reduces the invasion and metastasis of tumor cells.

OBJECTIVE To design and construct eukaryocyte expression vector of SV40 virus large T antigen and induce its targeted expression in eukaryocyte.METHODS SV40 large T gene which excised intron was cloned by SOE(splicing by overlapping extension) and digested with restricted enzymes EcoR Ⅰ and BamH Ⅰ.By the same methods,we got the digested product of pEGFP-N1.After that,the two fragments were ligated to form SV40(TEGFP) by Ligation Kit,and sequenced by TaKaRa ABI Prism Terminator Cycle Sequence Kit.The reconstructed vector was transfected into primary cultured human fibroblast using a Lipofectin transfection method.At 48 h(after) transfection,the expression of SV40T was detected with PCR and RT-PCR using specific primer of T gene.(RESULTS) The restricted enzymes digested and sequencing results showed that SV40 large T gene had cloned into pEGFP-N1 vector successfully.The genome DNA and total RNA were isolated from the positive cells.With these samples,the specific 288 bp fragment was amplified using PCR and RT-PCR.CONCLUSIONS The recombinant plasmid SV40TEGFP will be a stable and valuable molecular tool for human eukaryocyte study.

Objectives To explore the influence of standardized treatment and technical flow improvement on the diagnosis and treatment of congenital diaphragmatic hernia (CDH) after prenatal diagnosis. Methods The clinical data of neonates diagnosed with CDH who were hospitalized in neonatal intensive care unit from January 2005 to August 2016 was analyzed retrospectively. Based on the start time of standardized treatment implementation, 93 cases of CDH hospitalized from January 2010 to August 2016 were divided into the intervention group while 15 cases of CDH hospitalized from January 2005 to December 2009 were divided into the control group. The survival rate and complications of clinical outcomes between two groups were compared. Results There was no significant difference in sex, gestational age, birth weight, Apgar score, and CDH position between two groups (P >0.05). The total survival rate was 81.7％ in the intervention group and 53.3％ in the control group, and there was statistical difference (P<0.05). The difference of mechanical ventilation mode and pneumothorax rate between two groups also had statistical difference (P >0.05). The median operation time in the control group was 4.5 h and the intervention group was 49.5 h. The postoperative survival rate was 61.5％ in the control group and 90.5％ in the intervention group, and the differences between two groups were significant (P >0.05). The first blood gas analysis of deaths cases in both groups showed that there were significant differences in pH and PCO2 values (P >0.05). Conclusions Optimization of the clinical management during perioperative period can improve the survival rate of CDH and reduce complications. However, the dead cases in the intervention group had more severe pulmonary hypoplasia.

Aged ; Coal Mining ; Electrocardiography ; Heart Rate ; physiology ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; physiopathology