We conducted the detection the Francisella spp.nucle acid from Hyalomma asiaticum asiaticum that main distribution is on railway line area from China-Kazakhstan border.The free-living ticks were collected and then identified by morphological and molecular methods.After species identification,they were detected by PCR targeting 16S rRNA and sdhA of Francisella spp.The amplified products were sequenced and the sequences was analyzed by using the Blast.A phylogenetic tree was constructed using MEGA 6 software.A total of 243 fleas were identified as H.asiaticum asiaticum.Only 35 samples were detected for Francisella spp.positive and the positive rate was 14.4％.Sequence analysis showed that two different sequences (seql and seq2) and all belong to Francisella-like endosymbionts (FLEs).Phylogenetic analyses showed that two FLEs were belong to the same cladd.This is first detection of FLEs nucleic acid from H.asiaticum Railway line area of China-Kazakhstan border.

Non-O1/O139 group of Vibrio cholerae can cause human acute diarrhea disease,while compared with the O1 and O139 groups;it often ignore the risk of the disease for human being.Therefore,we analyzed the molecular characteristics of 31 V.cholerae isolated from Yunnan Province.We used the agar disc diffusion method (K-B) to carry out the antibiotic sensitivity test;polymerase chain reaction (PCR) amplification for the detection of virulence gene;at the same time,all of strains were performed for pulse field gel electrophoresis (PFGE) and multilocus sequence typing (MLST).The drug sensitivity test showed that 67.74％ strains were resistant to rifampin,29.03％ resistant to nalidixic acid and cotrimoxazole,all of the isolates were sensitive to gentamicin and ciprofloxacin;PCR results showed that all strains had the ompW gene,87.10％ strains had hly gene,25.81％ strains had rstREl tor,16.13％ strains had rstRClassical and tcpAEl tor,while CT rfbO1 and rfbO139 gene were negative;PFGE results showed that 31 strains had a trend of discrete height,the same PFGE identity pattern was not nearly found;for the analysis of MLST,we found the one new alleles of gyrB,four new alleles of mdh gene,six new alleles of metE gene,two new alleles of pntA,three new alleles of purM and four new alleles of pyrC gene.After permutation and combination,we found 17 new ST types for V.cholerae(ST273-ST289).Non-O1/O139 group V.cholerae showed a high degree of diversity,while the non-O1/O139 group of V.cholerae in Yunnan Province has a certain geographical features,which enriched the existing molecular typing system of V.cholerae.

To identify the receptors for the outer membrane protein H (OmpH) of avian P.multocida,the membrane proteins of chicken embryo fibroblast (CEF) cells were separated by SDS-PAGE and analyzed by Ligand blot.The OmpH-binding protein was identified by MALDI-TOF mass spectrometry,and its distribution in the membrane proteins of different host esophageal mucosal cells was detected by Ligand blot,ELISA and immunofluorescence microscopy,respectively.Ligand blot analysis showed that a 49-kDa membrane protein of CEF cells bound to recombinant OmpH,and MALDI-TOF spectral results demonstrated that the OmpH-binding protein was ATP synthase β subunit.In addition,the OmpH receptor was present in the chicken and rabbit mucosal cell membranes,but was not detected in the bovine and swine mucosal cell membranes.The above results indicate that the OmpH receptor may be CEF cell-derived ATP synthase beta subunit.

The aim of this study is to analyze the evolutional and molecular characteristics of Hemagglutinin (HA),Neuraminidase(NA)and non-structural (NS) genes of H5N1 avian influenza virus (AIV) from sewage in live bird markets (LBMs) in Changsha,2014.Five hundred and one specimens were collected from environment in LBMs in Changsha,2014,and real-time RT-PCR was used for influenza A typing and subtyping (H5,H7 and H9) detection.Sequencing were used for the positive of single H5.The sequence homology of HA,NA and NS genes of the viruses were analyzed with the online Basic Local Alignment Search Tool (BLAST).The phylogenetic trees for HA,NA and NS genes and the ClustalW Multiple alignments of amino acids were constructed using MEGA 5 and BioEdit software,respectively.Results showed that of 501 environmental samples,177 (35.33 ％) samples were positive for influenza A viruses and H5 subtype.Eight H5N1 subtype AIV were confirmed by sequencing from the samples of the positive of single H5.Phylogenetic analysis indicated that most of HA genes of the H5N1 subtype AIV strains isolated in Changsha city were located in 2.3.2 and clustered into new subclade,and the most of NA and NS genes in this study were clustered into subclade 2.3.2.1b.QSG of the HA protein of the receptor binding site were found in these H5N1 viruses,and the characteristics was shown to be associated with increased affinity of HA to the glycan-receptors of AIV.In strains from this study,we did not found amino acid substitutions of the NA protein at H275Y and N295S,and sensitive to neuraminidases,and the high pathogenicity molecular characteristics of HA,NA and NS genes were showed in these viruses.In conclusion,molecular characteristics of the HA,NA and NS of these H5N1 subtype viruses in this study showed high pathogenicity,but that may not facilitate human infection.So,the prevalence and genetic evolution of this virus should be closely monitored.

We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.

The situation of the ongoing fifth epidemic (beginning October,2016) of human infection with avian influenza A (H7N9) virus are more serious than the first four ones.As of March 8,2017,the fifth epidemic reported 40.00％ of the cumulative cases.Recently,the pathogenic study reported two new human infections in Guangdong with an influenza A(H7N9)virus strain for which gene sequencing analysis revealed mutations in the haemagglutinin (HA) gene that resulted in the insertion of basic amino acids at the cleavage site of this protein,known to confer increased pathogenicity in chickens.However,most strains show no obvious difference in pathogenic characteristics compared with those detected during the first four waves.Although epidemiological studies showed that 3 clusters with limited human-to-human transmission have been investigated during the current wave,there are no indications of sustained person-to-person spread.Based on epidemic analysis and risk assessment results,it's likely that human infections with H7N9 virus will continue to occur in China,but epidemiological and pathogenic analysis suggested that it's unlikely to have a continued transmission of this virus.Profound public health significance would be presented by strengthening etiology and epidemiology studies of avian influenza A(H7N9) virus.

We analyzed the cost effectiveness of four different surgical methods (A,B,C and D) in treatment of hepatic echinococcosis.Totally 757 cases of hepatic echinococcosis surgery clinical data and cost information of hospitalized cases were collected from nine hospitals in Xinjiang during 2005-2013.The clinical effects,cost effectiveness ratio and incremental costeffectiveness ratio were analyzed and compared.The total complication rates were 15.8％,9.2％,0％ and 2.9％;the recurrence rates were 7.3％,6.2％,0％ and 0％;the cure rates were 77.8％,84.6％,100.0％ and 97.1％;the cost (RMB) were 11 947.3,18 543.6,25 510.7 and 18 877.4,cost-effectiveness ratio were 153.6,219.1,255.1 and 194.3,incremental cost-effectiveness ratio /C//E were 964.4,610.4 and 358.0 respectively for group A,B,C and D.The results after adjusting of price factor were consistent with original cost effectiveness analysis.The complete resection of inner and outer capsule (D) is the most effective and economical way of clinical operation and may worthy be promoted in treatment of hepatic echinococcosis.

We investigated serotype and resistance of Salmonella during pig slaughtering in Shandong Province,China,providing basic data for the risk assessment of Salmonella and for guiding the clinical medication.We used rapid classification kit to identify Salmonella serotype,adopted broth microdilution method to detect the resistance of 13 kinds drugs belong to 8 categories.Result showed that the identified 9 kinds of serotype were mainly S.derby and S.typhimurium.The resistance to 13 kinds drugs of 298 Salmonella were different.The higher percentage of tetracycline drugs as Doxycycline(DOX) and Tetracycline(TE) were 97.99％ and 80.20％,respectively,which was most sensitive to Colistine E.The resistant rate was only 2.01 ％,following by Amoxicillin/Clavulanic acid and Ofloxacin which were 2.35 ％ and 4.03％,and the multiple resistant rate was 81.88％.TE-DOX was the regnant drug-resistant spectrum.In conclusion,the predominant serotype of Salmonella in links of pig slaughtering in Shandong Province is S.derby,and resistance is different to the different drugs.The drug resistance of different slaughter links exist significant differences,multiple drug resistance is serious,and drug-resistant spectrum are varied.

Tuberculosis (TB) remains an enormous health burden worldwide.To date,Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the unique anti-TB vaccine available for humans,which provides an important but limited protection from the Mycobacterium tuberculosis (Mtb) infection.It is therefore an urgent need to develop better vaccines and vaccination strategies to prevent the spread of Mtb infection.Heterologous prime-boost vaccination strategies using both BCG and novel anti-TB vaccines have been demonstrated to induce robust immune responses than BCG alone.We have previously demonstrated that a recombinant adenoviral vector Ad5-CEAB co-expressing CFP10,ESAT6,Ag85A and Ag85B of Mtb was able to induce robust antigen-specific immune responses in mice.In the present study,we examined immunological effects of Ad5-CEAB in the mice primed with BCG and boosted with a single dose of the virus via an intranasal route.Results demonstrated that this vaccination strategy could effectively induce strong antigen-specific mucosal and humoral immune responses.These immune responses were characterized with an increased productions of cytokines (IL-12 and IFN-γ),increased concentration of secretary IgA (sIgA) in bronchoalveolar lavage fluid (BALF) and serum IgG in mice in comparison with mice in BCG group.These data suggested that the regimen of BCG prime-single dose of Ad5-CEAB boosted strategy was novel for inducing antigen-specific immune responses in response to Mtb antigens in vivo,which warrants for further development of adenoviral-based vaccine against Mtb infection.

We investigated the carrying situation of Escherichia albertii from healthy people engaged in breeding and slaughtering poultry for a long time.We collected stool samples from people engaged in breeding and slaughtering poultry and other healthy people.After enriched with EC broth,eae-positive enrichment culture was directly streaked on MacConkey,and eaepositive lactose non-fermenting isolate was retained for further investigation.The 16S rDNA sequencing and multilocus sequence typing (MLST) were applied in the identification of E.albertii from suspected strains.Intimin subtypes and cdtB types of E.albertii strains were detected.Pulsed-field gel electrophoresis (PFGE) was used to detected genetic polymorphism of strains from this study and animal source ones.Results showed that two isolates were identified as E.albertii from 189 stools of people exposed to slaughtering chickens and ducks and one from 58 stools in control groups.No isolate was identified as E.albertii from 138 stools samples of people exposed to breeding poultry.Intimin subtypes of three isolates from stool samples were subtyped as sigma,iota 2,nu,and cdtB types were closely related to types Ⅱ/Ⅲ/Ⅴ.PFGE patterns of the three strains was distinguishable (＜80％ similarity),and appeared in different cluster with chickens,ducks and other sources of E.albertii strains.The rate of carrying E.albertii to a certain extent exist in healthy people engaged in slaughtering chickens and ducks,and the relationship between these strains and strains from poultry should be further investigated.