Objective To simulate the process of lag screw insertion on intact pelvises under guidance of conventional fluoroscopes or 2D and Iso-C3D computer-assisted navigations and evaluate the accuracy and practicability of the computer-assisted navigation. Methods Six dried intact adult pelvic specimens were selected and divided into three groups randomly. A total of 54 hollow screws were placed in bilateral pedicles of S1 and S2, anterior column of bilateral acetabulum, anterior column of bilateral acetabulum and pubic symphysis of intact adult dried pelvic specimens of three groups under guidance of conventional fluoroscopy, 2D and Iso-C3Dcomputer assisted navigations, respectively. The accuracy of the screw positions, the average operating time of each screw insertion and the average time of radiation exposure during the insertion of each screw were compared among three groups. Results There were significant differences in the accuracy of the screw positions, the average operating time and the average time of radiation exposure among three groups (P＜0.01). The navigation with Iso-C3D appeared to provide the highest accuracy and the shortest operating time of all guidance techniques. The mean operating time and the average time of radiation exposure of the conventional fluoroscope were the longest among three groups. The average time of radiation exposure of the 2D computer-assisted navigation was the shortest.Conclusions Iso-C3D computer-assisted navigation is the most accurate and expeditious means of all guidance techniques. The time of radiation exposure can be significantly reduced by both 2D and Iso-C3D computer-assisted navigations.

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.

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

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.

OBJECTIVE: To explore the possible mechanism of Huoxue Qianyang Formula (HXQYF), a compound traditional Chinese herbal medicine, in reversing the left ventricular hypertrophy (LVH) of spontaneous hypertensive rats (SHRs) by analyzing the expressions of mRNAs and proteins of proto-oncogenes c-fos and c-myc in left ventricular muscle. METHODS: The experimental study was carried out in SHRs, the sex- and age-matched Wistar-Kyoto (WKY) rats were served as normal control (n=5, normal saline 10 ml/kg daily). Twenty-five SHRs were randomly divided into five groups: untreated group (n=5, normal saline 10 ml/kg daily), high-dose HXQYF-treated group (n=5, 0.84 g/ml HXQYF, 10 ml/kg daily), medium-dose HXQYF-treated group (n=5, 0.42 g/ml HXQYF, 10 ml/kg daily), low-dose HXQYF-treated group (n=5, 0.21 g/ml HXQYF, 10 ml/kg daily) and cilazapril-treated group (n=5, 1 mg/ml cilazapril, 10 ml/kg daily). The drugs were intragastrically administered once daily for 14 weeks. The expressions of mRNAs and proteins of proto-oncogenes c-fos and c-myc in left ventricular muscle were detected separately by in situ hybridization histochemical method and immunohistochemical method. RESULTS: Compared with the normal control group, the expressions of mRNAs and proteins of proto-oncogenes c-fos and c-myc in left ventricular muscle were significantly increased in untreated group (P<0.01). After treatment, the expressions of c-fos and c-myc mRNAs in left ventricular muscle in HXQYF-treated groups were significantly down-regulated as compared with those of the untreated group (P<0.05). The expressions of c-myc protein were also significantly decreased in high- and medium-dose HXQYF-treated groups as compared with the untreated group (P<0.05), but it had no significant effects in protein expression of c-fos in the three HXQYF-treated groups. CONCLUSION: HXQYF can inhibit the expression of c-myc in ventricular hypertrophy tissue, which may be the mechanism in treating LVH of hypertension.

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

Angiostatin(AS) and endostatin(ES) are both potent endogenous angiogenesis inhibitors, and the combination of AS and ES has been shown to have synergistic antiangiogenic effects. Here we report the fusion protein AS-ES expressed in E. coli which has antiangiogenic effects. At first, AS and ES genes were cloned respectively through RT-PCR, then fusion gene was made through gene splicing ,finally pET-42 (b)/AS-ES expression plasmid was constructed and transduced in E. coli BL21 (DE3). Target protein was in form of inclusion body,the rate of expression was about 14%, and MW about 65KD. Western blotting assay showed expressed protein had specific immune reaction to both the antibodies of AS and ES. The expressed protein which was refolded and purified through heparin affinity chromatography had antiangiogenic effect to vessels on chicken embryo chorioallantoic membrane. The results show that fusion protein AS-ES was expressed successfully in E. coli, and the expressed protein,which was renatured and purified, had immuno-reactivity to anti-AS and anti-ES in Western blotting and angiogenesis inhibition activity.
Angiogenesis Inhibitors ; Angiostatins ; biosynthesis ; genetics ; isolation & purification ; pharmacology ; Animals ; Blotting, Western ; Chick Embryo ; Endostatins ; biosynthesis ; genetics ; isolation & purification ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; pharmacology ; Recombination, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective To study the genome molecular characteristics of Getah virus(DY0824)which isolated in Shandong province,2008 by molecular biology methods.Methods Reverse transcriptasepolymerase chain reaction(RT-PCR)was used to amplify the structural gene and 3'UTR fragments then the RT-PCR products were inserted into PGEM-T easy to be sequenced.Computer software was used to analyze the nucleotide and deduced amino acid sequence,and draw phylogenetic trees,including Clustal X1.83 and MegaAlign and Mega4.Results The capsid protein of DY0824 consists of 804 nucleotides,encoding 268 amino acids and the full-length of E2 protein is 1266 nucleotides,encoding 422 amino acids.The nucleotide homology of the capsid protein and the E2 protein with other strains were 95.4%-99.9%and 94.8%-99.5%,and the amino acid were 97.4%-100%and 97.6%-100%.The 3'UTR of the virus include 401 nucleotides and there are three repeat sequence elements.Conclusion Compared with the prototype virus,the Getah virus isolated in Shandong province had 7 amino acid differences in capsid protein genes and 10 amino acid differences in E protein genes.The 3'UTR region had multi-nucleotide changes.

Objective To analyze the genotype of Japaneso encephalitis virus (JEV) strains isola-ted in 2004 from mosquitoes collected in Bazhong city, Sichuan province of China, and the characters of amino acid in the PrM and E gene. Methods The isolated virus strains from mosquitoes were identified by biological, serological and molecular biology. PrM and E segments of the isolated JEV were amplified by RT-PCR, the PCR products were purified and sequenced. Multiple alignment, phylogenetic and amino acid (AA) analysis were carried out by Clustal X (1.8) , MEGA4 and GENEDOC (3.2) . Results The total of 4688 mosquitoes were collected including Armigeres and Culex. Six isolates were identified be-longing to genotype 1 JEV. The comparison between new genotype 1 JEV strains and live attenuated vaccine strain SA14-14-2 in PrM and E gene showed that total 3 sites amino acid differences in PrM gene and 14 sites in E gene, respectively. Three sites (PrM2, 64 and 65 ) in PrM protein and four sites (E129, 222,327 and 366) in E protein were only belonging to genotype 1 JEV. Conclusion The new isolated JEV strains in Sichuan province belong to genotype 1. It suggests that the vaccine strain SA14-14-2 currently used for preventing Japanese encephalitis is able to protect people against JEV, although in the segments of it had some amino acid differences between vaccine strain and the epidemic genotype 1 JEV strains in PrM and E gene.