Objective To elucidate the molecular basis on the differences of infectivity in infected cells between Sindbis virus(SINV:YN87448 virus)and Sindbis like virus(SINLV:XJ-160 virus).Methods Compare the E(glycoprotein)gene sequence and secondary structure of YN87448 virus and XJ-160 virus by bioinformatics analysis.Analyze the contribution of E gene to the biological differences between SINV and SINLV by constructing recombinant virus.Results By bioinformatics analysis,YN87448 virus and XJ-160 virus have the same genomic structure,which has 11 717 nt and 11 626 nt respectively.There are 82 amino acid differences between E gene of these two viruses,and showed scattered distribution.The main peak is basically the same for the hydrophobic of the E gene protein,but in some region existing small differences.The recombinant virus which exchanged the E gene of XJ-160 virus with YN87448 virus totally showed the biological character of YN87448 virus,either in the showing time of CPE,plaque forming time and plaque diameter,or in expression of functional proteins.Conclusion E gene plays a major role in the differences of infectivity in infected cells between SINV and SINLV,this result provide the molecular biological evidences for elucidating the biological differences between SINV and SINLV.

Objective To explore the relationship between insluin resistance(IR)and plasma fibrinogen, blood lipids levels in patients with gestational diabetes mellitus (GMD).Methods Totally 127cases pregnant women in their 24 -28 gestational weeks were divided into GDM group(n =37),gestational impaired glucose tolerance test (GIGT)group(n =30)and normal glucose tolerance(NGT)group(n =60)based on the American Diabetes Associ-ation(ADA)2009 GDMdiagnostic standards.All selected cases's height,weight,fasting blood glucose (FBG),fasting insulin (FIN),blood lipids and fibrinogen(FIB)level were tested,and their body mass index(BMI)and IR index (HOMA -IR)were calculated.Results During the mid -trimester,the levels of BMI,FIB,HOMA -IR,FBG,FIN, triglyceride (TG),cholesterol(TC),low density lipoprotein -cholesterol (LDL -C)in the GDM group were signifi-cantly higher than the GIGT and NGT groups.However,high density lipoprotein -cholesterol (HDL -C)was signifi-cantly lower (F =69.29,98.15,70.32、141.43、61.92,106.06,157.06,90.59,25.25,all P <0.01).FIB was sig-nificantly positively correlated with BMI,FBG,FIN,HOMA -IR,TC,TG,LDL - C (r =0.441,0.682,0.506, 0.599,0.493,0.591,0.633,all P <0.01 ),and negatively correlated with HDL -C(r =-0.419,P <0.01 ).Multiple stepwise regression analysis indicated that BMI,LDL -C,FBG and TG were independent risk factors affect-ing FIB(t =7.683,2.678,3.184,3.844,all P <0.01);FIB,BMI,FBG,FIN and TC were independent risk factors influencing HOMA -IR(t =68.784,3.201,29.702,58.100,2.904,all P <0.01).Conclusion Pregnant women have IR and blood lipid disorders during the mid -trimester,the overexpression of FIB is closely related to it.

Objective To investigate the effect of xenon intervention on delayed neuropsychologic sequelae (DNS)in acute carbon monoxide(CO)poisoning.Method Adult Wistar rats were randomly divided into sustained group,early intervention group,and control group.CO(150 ml/kg)was infused by intraperitoneal injection to produce DNS model.In sustained intervention group(S-group),xenon(150 ml/kg/d)was infilsed by intraperitoneal injection for 2 weeks;in control group(C-group),xenon was replaced by equal volume air;and in early intervention group(E-tvoup),xenon(150 ml/kg/d)was,employed in the first 3 days and air(150 ml/kg/d)was substituted for xenon in the following days until 2 weeks after CO poisoning.Morris maze test was used to evaluate the intelligence of rats.The long-term potentiation(LTP)of hippocampus Was detected by neuroelectricity recording.The apoptosis rates in brain was detected by TUNEL staining.The data were expressed as(x±s)and analyzed with student's test and analysis of variance.A P value less than 0.05 indicated statisfical significance.Results After exposure to CO,poisoned rats showed intelligence decline,demyeliation ofwater matler and cell apoptosis increased,which were consistent with DNS.In S-group and E-group,the rates of DNS and apoptosis were significantly lower than those in C-group,whereas the rote of LTP in S-group and E-group Was significantly higher than those in C-group.Conclusions Early xenon intervention can effectively decrease the rates of DNS occurred after acute CO poisoning.

Objective To investigate whether the characteristics of pathogens mediated by Wolbachia can interfere with Japanese encephalitis virus (JEV) replication and explore the regulatory role of Wolbachia on JEV replication transmitted by Culex mosquitoes. Methods Real-time fluorescence quantitative PCR (qPCR) and RNA fluorescence in situ hybridization (RNA-FISH) were used to detect Wolbachia density in Aa23 (naturally infected with Wolbachia) Aedes albopictus cell and negative control Aa23T Aedes albopictus cells (Wolbachia infection was removed by tetracycline treatment). The plaque assay was conducted to measure the viral titers and cytopathic effect (CPE) in Aa23T and Aa23 cells on days 1 to 8 after JEV (P3 strain) infection. Results qPCR and RNA-FISH results consistently showed that the symbiosis of Wolbachia was negative in Aa23T cell. In Aa23 cells, the copy number of the WSP gene of Wolbachia and the fluorescence signal intensity targeting Wolbachia 16S rDNA increased with cell growth time. In response to JEV infection, Wolbachia prolonged the CPE in viral infected Aa23 cells, which compared to infected Aa23T cells. The plaque assay result has obviously showed that JEV titer in Aa23 cells (106 PFU/mL) was significantly lower than that in Aa23T control cells (108 PFU/mL). Conclusions Wolbachia significantly delays CPE of JEV on cells and inhibits JEV replication in A. albopictus cells. To the best of our knowledge, this is the first report that Wolbachia strongly inhibits JEV infection in mosquito cells. It revealed the role of Wolbachia on inhibition of the viruses that transmitted by Culex mosquitoes. In particular, it provides important experimental data and theoretical basis for application of Wolbachia-based mosquito control technology in prevention and control of JEV.

To explore the spatial distribution mechanism of Japanese encephalitis virus (JEV), PhyML v3.0 was used to build phylogenetic tree using JEV sequences in the dataset. PAUP v4.0 and Migrapyhla softz ware were then used to analyze the migration events. The results showed that a total of 95 migration events were observed during the dispersal of JEV throughout Asia. Further analysis revealed that Thailand, and several Chinese provinces (including Shandong, Shanghai, Sichuan and Yunnan), were the main migration sources of JEV. JEV spread from these migration sources as follows: from Thailand to Australia, Cambodia, Tibet and India; from Shanghai to eastern coastal Asian regions and Yunnan; from Shandong to Korea, Zhejiang, Hubei, Shanxi and Liaoning; from Sichuan mainly to inland regions of China, as well as Vietnam and Japan; and from Yunnan to Zhejiang. This study indicated that frequent migration events occurred during the dispersal of JEV in the Asia and Pacific regions, and that Thailand, Shandong, Shanghai, Sichuan and Yunnan were the sources of JEV dispersal.
Asia ; epidemiology ; China ; epidemiology ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; physiology ; Encephalitis, Japanese ; epidemiology ; transmission ; virology ; Phylogeny

To evaluate the prevalence of mosquito-borne viruses in Manshi and Ruili (Yunnan Province, China), we collected 2 149 mosquitoes (17 species) in August 2010. Virus isolation was undertaken by the cul- ture of baby hamster kidney cells (BHK-21 cells). Two virus-like isolates were obtained: DHL10M117 was isolated from collected in Mangshi; DHL10M110 was obtained from Anopheles vagus collected in Rui- li. Both isolates caused cytopathic effects,illness and death in suckling mice inoculated with these isolates via the intracerebral route. Two positive amplicons, 702-bp from the S segment and 456-bp from the M segment,were obtained using reverse transcription-polymerase chain reaction using primers specific for the Akabane virus (AKV). Phylogenetic analysis suggested that these two virus stains had a distant relation- ship with AKVs from Kenya and Australia,but were genetically close to those from Japan,South Korea, and Taiwan. However,they were separate from other Asian strains and grouped into a small branch. The highest nucleotide and amino-acid sequence identity of the S segment was found with the CY-77 strain from Taiwan (96.6% and 99.6% for DHL10M117 and 96.7% and 100% for DHL10M110,respectively). Com- parison of the M segment showed they shared the highest amino acid identity with CY-77 (99.6% and 100%, respectively), whereas the highest nucleotide identity was found with the Iriki strain from Japan (99.6% and 100%, respectively). Compared with the MP496 strain from Kenya,they displayed lower lev- els of sequence homology, at 69.7% and 70.0% for nucleotide sequences of the two loci,and 91. 0% for a- mino acids. Our results identified that DHL10M117 and DHL10M110 were strains of AKV,and provided molecular biological evidence for the existence of AKV in Yunnan Province. These AKV strains that are circulating in Yunnan Province share a close genetic relationship with strains from the rest of Asia. Culex tritaeniorhynchus and Anopheles vagus may serve as transmission vectors.
Amino Acid Sequence ; Animals ; Anopheles ; virology ; Base Sequence ; Bunyaviridae Infections ; virology ; China ; Cricetinae ; Female ; Humans ; Insect Vectors ; virology ; Male ; Mice ; Orthobunyavirus ; classification ; genetics ; isolation & purification ; physiology ; Phylogeny ; Sequence Homology ; Viral Proteins ; chemistry ; genetics

Arboviruses represent a serious problem to public health and agriculture worldwide.Fast,accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation.We developed a cost-effective,rapid,and highly sensitive one-step triplex RT-PCR enzyme hybridizationassay for simultaneous detections of Japanese Encephallitis virus (JEV,Flaviviridae)Getah virus (GETV,Togaviridae),and Tahyna virus (TAHV,Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction.The analytical sensitivity of this assay was 1 PFU/mL for JEV,10PFU/mL for GETV,and 10 PFU/mL for TAHV.This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods.When “triplex RT-PCR enzyme hybridization” was applied to 29 cerebrospinal fluid(CSF)samples that were JEV-positive by normal RT-PCR assay,all samples were strongly positive for JEV,but negative for GETV and TAHV,demonstrating a good sensitivity,specificity,and performance at CSF specimen detection.

0.05)). The Val/Leu genotype and Leu allele frequencies in subjects without MI were significantly higher than that in subjects with MI (P

Objective To explore the relationship of serum ferritin with insluin resistance(IR)and inci-dence of fatty liver in adults with simple obesity.Methods According to body mass index (BMI),78 adults with simple obesity were divided into three groups,including 34 cases in Ⅰ degree obesity group,30 cases in Ⅱ degree obesity group and 14 cases in Ⅲ degree obesity group,and 35 adults with simple overweight and 67 healthy adults were also enrolled as control group.The height,weight,waistline,hip circumference,incidence of fatty liver,fasting blood glucose (FBG),fasting insulin (FIN),blood lipids and ferritin(SF)level were tested,and the body mass index (BMI),WHR and IR index (HOMA -IR)were calculated.Results When BMI increased,levels of SF,HOMA -IR, FBG,FIN,triglyceride (TG),cholesterol (TC),low density lipoprotein -cholesterol (LDL -C)were gradually increased(F =378.92,12.01,55.50,123.96,90.09,127.65,23.81,all P <0.01).However,high density lipopro-tein -cholesterol (HDL -C)was gradually degraded(F =114.56,P <0.01).The incidence of fatty liver had signifi-cant differences in Ⅰ,Ⅱ,Ⅲ degree obesity groups((t =24.872,7.885,all P <0.01),and with the degree of obesity increased,the rate of fatty liver increased significantly.SF range interquartile and level in fatty liver group was signifi-cantly greater than the non fatty liver group,the difference was statistically significant (t =22.99,P <0.01).SF was positively correlated with BMI,WRH,FBG,FIN,HOMA -IR,TC,TG,LDL -C (r =0.863,0.719,0.789,0.703, 0.617,0.785,0.717,0.771,all P <0.01),and negatively correlated with HDL -C(r =-0.530,P <0.01).Multi-ple stepwise regression analysis indicated that BMI,FIN,LDL -C,HOMA -IR were independent risk factors affecting SF(t =4.646,2.595,-5.073,6.666,all P <0.01).SF,BMI,FBG,FIN,LDL -C were independent risk factors influencing HOMA -IR(t =6.535,3.254,18.827,61.227,2.154,all P <0.05).Conclusion Simple obesity adults had obvious blood lipid disorders and IR,easy to complicated with fatty liver,the incidence of fatty liver associ-ated with severity of obesity,the overexpression of SF is closely related to it.

Objective To identify the Banna viruses isolated in northwestern part of Yunnan prov-ince in order to make the difference clear between the isolates and other Banna viruses isolated in other parts of Yunnan. Methods Three isolates of Banna vires isolated in 2005 and 2006 were identified by morpholo-gy, RNA-PAGE profile and molecular biologic method. Nueleotide and amino acid sequences of segment 12 of the 3 isolates were sequenced and analyzed. Results Three Banna viruses were isolated from mosquitoes collected in northwestern part of Yunnan during 2005 and 2006. Electron microscopy study showed that they are spherical with a diameter of 70 nm, no envelope but two layers of eapsid. It was found that the genome of the 3 isolates composes of 12 segments presenting band profile of 6-6 in RNA-PAGE. Nueleotide acid se-quence analysis about segment 12 showed that the identity was 99% between the 3 new isolates, 98% and 90% between the 3 isolates and the strains isolated in other parts of Yunnan, China and Indonesia, respec-tively. Phylogenetie analysis based on segment 12 gene showed that 3 new isolates clnstered in the same branch with the viruses isolated in other parts of Yunnan. The same difference of amino acids was found between Banna viruses isolated in China and Indonesia strains in the analysis of segment 12. Conclusion Banna virus strains were firstly isolated from mosquitoes collected in northwestern part of Yunnan province. Nueleotide acid sequence analysis of the 3 new isolates showed higher identity with strains isolated in other parts of Yunnan.