To identify the isolated suspicious strain of Campylobacter jejuni from the blood of bacteremia patient in Guizhou Province ,China ,conventional and molecular techniques (specific mPCR and NAP-mPCR) were used to identify suspi-cious bacteria strains .Results showed that Campylobacter jejuni suspicious colonies were cultured in bacteremia patient blood samples .The strain was identified as Campylobacter jejuni ssp . jejuni by conventional tests and was identified as Campy-lobacter jejuni by genus specific mPCR .Then the strain was classified as Campylobacter jejuni ssp . jejuni by subspecies NAP-mPCR .The strain was identified as Campylobacter jejuni ssp .jejuni isolated from the blood of bacteremia patient and Campylobacter jejuni can be identified subspecies by NAP-mPCR .

Objective To study the effect of functional mandible shift on mandible development in pubertal rats.Methods Forty-eight male Sprague-Dawley rats at the age of 28 d were randomly divided into experimental group (n=24) and control group (n=24).Maxillary occlusal splints were designed to simulate mandible shift in experimental group.Six rats in each group were killed after 7,14,21,and 28 days,respectively.Right and left mandibles were removed for X-ray radiography and localizing points.Distance between the points was measured with an image analyzer.Results Fourteen days from experiment,the length parameters of the non-shifted mandible(Cd-Me,Co-Me,Go-Me)were significantly higher than those of the shifted mandible length in rats of experimental group(P

BACKGROUND:Whether the functional mandibular deviation wil cause osteal mandibular deviation and whether the mandibular growth pattern wil change are stil controversial. Vascular endothelial growth factor can increase the endothelial permeability, stimulate endothelial cel mitosis and promote angiogenesis and regulate the formation of bone. OBJECTIVE:To investigate the effect of functional mandibular deviation on the expression levels of vascular endothelial growth factor in condylar cartilage in growing rats. METHODS:Sixty four-week-old male Sprague-Dawley rats were randomly divided into control group (n=20) and experimental group (n=40). Each animal in experimental group received a nichrome appliance which was designed by our team. Lower mandible suffered from left functional deviation (2.1±0.3) mm to simulate functional mandibular asymmetry. RESULTS AND CONCLUSION:The number of vascular endothelial growth factor-positive cels in the sagittal posterior deviation region of rats with functional mandibular deviation was less than that in the non-deviation region at 7-28 days after model establishment. The number of vascular endothelial growth factor-positive cels was less in the sagittal central and coronal central deviation region than in the non-deviation region in rats with functional mandibular deviation at 14-28 days after model establishment, but was close to the control group. The number of vascular endothelial growth factor-positive cels was less in the coronal lateral deviation region than in the non-deviation region and control group in rats with functional mandibular deviation at 14-28 days after model establishment. These results indicated that after the mandible functional lateral deviation in adolescent rats, the expression of vascular endothelial growth factor was different in bilateral condylar cartilage, caused the alterations in osteoblast activity in cartilage, and possibly affected the normal growth and development of rat mandible, suggesting that functional mandibular deviation should be corrected as early as possible.

Objective To evaluate the inhibitory effects of shRNAs targeting genes encoding viral capsomeres VP1-VP4 on enterovirus 71 (EV71) replication when used alone or in combination.Methods Short hairpin RNAs (shRNAs) targeting genes encoding VP1-VP4 protein of EV71 were designed and then respectively inserted into lentiviral vector pLKD-CMV-GFP-U6 to construct the recombinant plasmids.The expression plasmids together with psPAX2 and pMD2.G were transfected into 293T cells to induce the expression of recombinant lentiviruses,which were collected on the third day after transfection.The titers of recombined lentiviruses were determined by real-time PCR.The effects of shRNAs used alone or in combination on the expression of EV71 at mRNA and protein levels were respectively detected by real-time PCR and Western blot.Results The inhibitory effects of shRNAs on EV71 replication showed no significant differences among various strains (isolated from fatal cases,severe cases,mild cases and FY0805) (P＞0.05).Their inhibition rates were 51.6％ (sh-VP1-1),85.1％ (sh-VP1-2),76.4％ (sh-VP1-3),57.5％ (sh-VP2-1),81.4％ (sh-VP2-2),79.5％ (sh-VP2-3),68.9％ (sh-VP3) and 56.7％ (sh-VP4) respectively,and they were in a dosage dependent manner.sh-VP1-2 in combination with sh-VP2-2 showed the highest inhibition rate reaching up to 96.6％.Moreover,shRNAs used in combination showed better effects than any one used alone even at double dosage.Conclusion All shRNAs targeting viral capsid VP1-VP4 genes showed inhibitory effects on EV71 replication with inhibition rates over 50％ and the effects could be strengthened when using shRNAs in combination.

Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68％) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90％-96.72％ and 99.45％-100％.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.

Objective To isolate and identify the Dengue virus(DEN)from the sera of patients with unknown fever in summer and autumn in Dushan and Xingyi areas of Guizhou province,China.Methods From June 2005 to October 2005.356 blood samples were collected from patients with unknown fever in Dushan and Xingyi areas of Guizhou province.The serum samples were inoculated on the C6/36 cell monolayers.After three blind passages,the cytopathie effect(CPE)Was observed.Identification of DEN antigen was earried out by indirect immunofluorescence assay(IFA)with the monoclonal antibody(McAb)against DENl-4 virus.The total RNA was extracted from the serum and tested by RT-PCR with the universal primer for DEN NS region.And determination of the RNA sequenee Was performed,and phylogenetic analysis was carried out.Results Three serum samples caused CPE and were proved as DEN2 positive by IFA,RT-PCR and senquence determination.The phylogenetic tree analysis showed that the isolated virus strains had the closest relations with the systemic evolution of the strains DEN2-43 and DEN2-44.Conclusion DEN infections exist in the population of Dushan and Xingyi of Guizhou,China.

Eight patients with suspected cases of C .jejuni were etiologically diagnosed and analyzed in this study to pro-vide scientific basis for the confirmation of the cases of human campylobacteriosis in Guizhou Province ,China .Blood or feces of 8 suspected patients were employed to isolate bacteria strains .Conventional and multi-PCR techniques were applied to identify suspicious bacteria strains .The C .jejuni strains were analyzed by using Pulsed Field Gel Electrophoresis (PFGE) .Suspicious strains of C .jejuni were isolated from all the 8 suspected patients of campylobacteriosis and anticipated genes fragment were detected with multi-PCR .With the digestion of restriction enzyme SmaI ,the 8 C .jejuni strains were divided into 7 PFGE pat-terns with 7-10 DNA bands .Cluster analysis showed that the gross similarity of 8 strains of C . jejuni was more than 50% . The similarity of PFGE patterns between strain GZ201004 and GZ201005 from diarrhea patients was as high as 100% ,while the similarity of strain GZ201201 and GZ201007 was 66 .7% .Moreover ,C . jejuni were detected from all the suspected pa-tients of campylobacteriosis .PFGE results indicated that strains GZ201004 and GZ201005 were from the same source ,while all the 8 isolates showed PFGE polymorphism .

Objective To confirm the death of a child injured by a dog was due to rabies and to understand the molecular biologic features of rabies virus in Kaili,Guizhou province.Methods Brain tissue samples of patient and dog were collected to detect the rabies virus by direct immunofluorescence assay (DFA) and RT-nested PCR assay.Homology and phylogenetic tree were analyzed based on the whole nucleotide and deduced amino acid sequence of N gene of rabies virus followed by molecular epidemiological analysis.Results Both the human and dog brain tissue samples were confirmed positive by DFA and RT-nested PCR assay.The homology analysis of N gene sequences among GZH,GZD and other epidemic and vaccine rabies strains isolated from other provinces and other countries indicated that the detected samples shared the highest homology with the strain detected in Anlong prefecture in Guizhou in the year of 2006,and the homology between GZH and GZD was as high as 100％.Besides,among the vaccine strains,GZH and GZD showed the highest homology with strain CNT.Phylogenetic analysis indicated that the two samples were very close and belonged to genetype 1 lyssavirus,with the closest relationship between samples reported in Guizhou and Beijing.Conclusion It was confirmed on the viral molecular level that both the human and dog in Kaili were suffered from rabies,and the pathogens were genetype 1 lyssavirus.The prevalent strains in Kaili city was probably imported from other prefectures of Guizhou province,suggesting that prevention and control measures on rabies in Guizhou province should be strengthened.

To study the characteristic of glycoprotein gene sequence of rabies virus in Guizhou Province in recent years and provide scientific basis for effective control and prevention of rabies,RT-PCR assay were used to detect human and dog brain tissues derived from different prefectures of Guizhou.The amplification products were sequenced and analyzed with bioinformatics software.The results showed that the full-length G gene sequences of 8 positive samples were obtained by RT-PCR amplification,sequencing and splicing.The homology of eight G gene sequences from Guizhou Province were between 87.4％ -99.9％ and 83.3.％- 100％ on nucleotide and deduced amino acid level,respectively,and the highest homology were found with the genotype 1 strains ( 86.5 ％- 87.0％ for nucleotide and 83.3 ％- 100 ％ for amino acid) among genotype 1- 7 representative strains.Besides,phylogenetic analysis based on the G gene indicated that the relationship of 8 strains derived from Guizhou were closest to genotype 1 Lyssavirus,and the strains of Guizhou were very close to the strains come from Hubei,Hunan,Anhui,Guangxi,Jiangsu provinces and Shanghai,except for GZ01 and GZ09.Moreover,GZ09 were evolutionarily closed to the strains come from Malaysia and Thailand,while the remaining sequences were closed to the strain of Indonesia.This study confirmed on the G gene level that rabies virus epidemic strains circulated in Guizhou Province in recent years belonged to rabies virus genotype 1 and the evolutionary relationship with reported strains come from different provinces of China and different countries were revealed in this study.It will provide scientific basis for effective control and prevention of human and animal rabies in Guizhou Province.

We identified the routes of infection and evaluate the effect of disinfection on the field of an outbreak of an-thrax in a village of Wengan County ,Guizhou ,China ,thus trying to provide with basis for the implementation of policies for pre-vention and control of anthrax .The authors gathered the cases information by searching and interviewing the targeted persons house by house ,and reviewed the medical records in hospitals .The samples including patient’s discharging fluid ,residues of died horse ,and soil from the places where the villagers dismembered horse were gathered and cultured for Bacillus anthracis . The technique of multiple-locus variable-number tandem repeat analysis (MLVA-8) was applied for revealing the genetic rela-tionships among those isolated Bacillus anthracis strains .Five cases of cutaneous anthrax occurred in the outbreak and the total attack rate was 7 .58% (5/66) among those contactors evolved in the activity of carrying ,dismembering ,washing ,chopping and eating the died horse .The attack rate was 100% (3/3) for those who carried ,dismembered ,washed ,chopped and ate that horse ,100% (1/1) for those who carried ,dismembered and ate ,and 7 .14% (1/14) for those who washed ,chopped and ate . The 25% (1/4) of the samples of discharging fluid from the cases with cutaneous anthrax were positive .After disinfection , 15 .38% (4/26) of the soil samples retained positive .The genetic similarity was 100% among the 5 isolate strains .The results suggested that the outbreak of anthrax in villagers occurred through the activities of carrying ,dismembering ,washing and chop-ping the died horse .Strengthening the risk communication and disinfection of the dismemberment places should be the crucial strategies to prevent and control anthrax epidemics in Guizhou in the future .