Key Laboratory of Jilin Province for Zoonosis Prevention and Control

Changchun, China

Key Laboratory of Jilin Province for Zoonosis Prevention and Control

Changchun, China
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Liu J.,Academy of Military Medical science | Liu J.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Wang,Jilin University | Liu Y.J.,Changchun University of Chinese Medicine | And 12 more authors.
Letters in Applied Microbiology | Year: 2012

Aim: To prepare enteropathogenic Escherichia coli (EPEC) E2348/69 ghosts and investigate whether immunization with EPEC bacterial ghosts can elicit protective immune responses. Methods and Results: A recombinant plasmid with double λPL/PR-cI857 temperature-sensitive regulatory cassettes was constructed. The lysis gene E and/or the staphylococcal nuclease A (SNA) gene were separately inserted downstream of the two regulatory cassettes to construct the lysis plasmids pBV220::E and pBV220::E::CI-P-SNA. An EPEC reference strain E2348/69 (serotype O127:H6) was transformed with the lysis plasmids to produce EPEC ghosts. Mice injected with bacterial ghosts EGE (EPEC ghosts produced using lysis protein E) or EGES (EPEC ghosts produced using a combination of lysis protein E and SNA) gained weight normally and showed no clinical signs of disease. Vaccination trials showed that mice immunized with EGE or EGES were significantly protected against subsequent challenge with the wild-type virulent parent strain, EPEC E2348/69 (42/50 and 45/50 survival, respectively); in contrast, none of the 30 control mice survived. Conclusions: Immunization with EPEC ghosts can elicit protective immune responses in BALB/c mice. Significance and Impact of the Study: EPEC ghosts may represent a promising new approach for vaccination against EPEC infection. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.


Zhang Y.,Academy of Military Medical science | Guo Z.,Academy of Military Medical science | Wang Z.,Academy of Military Medical science | Fu Y.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | And 3 more authors.
Journal of Aerosol Science | Year: 2017

Influenza A virus has caused intermittent pandemics with potentially devastating consequences in human populations. Under natural conditions, influenza virus is mainly spread person-to-person through the air; however, in studies of influenza virology, the virus is typically intranasally instilled in the form of large liquid droplets. The dynamics of influenza virus infection and real-time progression of respiratory tract infection are still poorly understood, partly due to a lack of available efficient replication-competent viruses that stably express a reporter gene. To address this limitation, we constructed a replication-competent influenza A virus carrying a Gaussia luciferase reporter gene in the NA segment of the viral genome (IAV-Gluc). The recombinant virus (IAV-Gluc) stably expressed Gaussia luciferase, and the viral load in lungs was proportional to the fluorescence intensity. Although IAV-Gluc was less virulent than wild-type virus (PR8), it efficiently infected and replicated within murine lungs and was pathogenic in mice. After challenging guinea pigs with the equivalent doses of virus using two different methods, namely, intranasal (IN) inoculation and aerosol exposure (AR), it was found that the animals subjected to IN inoculation showed greater virus deposition in the lungs (12.27%) than those subjected to AR (1.34%), although the latter group exhibited more rapid viral replication within 48 h. Combining the results of live-animal imaging and traditional techniques used to measure viral titer, we identified additional differences between IN inoculation and AR. For example, IN inoculation caused the virus to distribute in a punctate pattern throughout the lungs, whereas virus delivered through AR showed an even distribution pattern, which increased the efficiency of viral replication. Indeed, the replication efficiency of the AR group was 11,073-fold higher than the original deposition amount after 48 h, whereas that of the IN group was only 15.4-fold higher than the original deposition amount after 24 h. © 2017 Elsevier Ltd


Li S.Y.,Academy of Military Medical science | Li S.Y.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Sha Z.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Sha Z.,Jilin University | And 11 more authors.
Current Microbiology | Year: 2017

Enterotoxigenic Escherichia coli (ETEC) is a significant cause of infectious diarrhea in animals. In this study, yeast surface display technology was employed to investigate the effects of ETEC enterotoxin fusion protein on the intestinal flora and mucosal immunity of rats. ETEC estA, estB, and eltAB (heat-labile and heat-stable toxins) were expressed on the surface of yeast. Rats were divided into normal saline, yeast and display yeast groups. Fecal and jejunal content samples were collected on the 7th, 14th, and 21st days. Rats were then fed ETEC for 3 days before again collecting these samples. Levels of SIgA, IL-2, IL-4, IFN-γ, and microbial population density and diversity were documented by ELISA, T-RFLP and real-time PCR. The results demonstrated that estA, estB, and eltAB fusion proteins were expressed on the surface of yeast. Following ETEC challenge, levels of SIgA, IL-2, IL-4, IFN-γ, and, the numbers and variety of intestinal microbes were significantly increased in rats receiving display yeast and yeast. These factors were significantly decreased in rats given normal saline and yeast. Our results indicate that display yeast and yeast can increase the number and diversity of intestinal microbes in rats and improve intestinal immune function. After ETEC challenge, the display yeast can better maintain the balance of intestinal bacteria and mucosal immunity. © 2017, Springer Science+Business Media New York.


Ni M.,Beijing Institute of Radiation Medicine | Chen C.,Capital Medical University | Chen C.,Beijing Key Laboratory of Emerging Infectious Diseases | Qian J.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | And 25 more authors.
Nature Microbiology | Year: 2016

Since 2013, West Africa has encountered the largest Ebola virus (EBOV) disease outbreak on record, and Sierra Leone is the worst-affected country, with nearly half of the infections. By means of next-generation sequencing and phylogeographic analysis, the epidemiology and transmission of EBOV have been well elucidated. However, the intra-host dynamics that mainly reflect viral-host interactions still need to be studied. Here, we show a total of 710 intra-host single nucleotide variations (iSNVs) from deep-sequenced samples from EBOV-infected patients, through a well-tailored bioinformatics pipeline. We present a comprehensive distribution of iSNVs during this outbreak and along the EBOV genome. Analyses of iSNV and its allele frequency reveal that VP40 is the most conserved gene during this outbreak, and thus it would be an ideal therapeutic target. In the co-occurring iSNV network, varied iSNV sites present different selection features. Intriguingly, the T-to-C substitutions at the 3′-UTR of the nucleoprotein (NP; positions 3008 and 3011), observed in many patients, result in the upregulation of the transcription of NP through an Ebola mini-genome reporting system. Additionally, no iSNV enrichment within B-cell epitopes of GP has been observed. © 2016 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.


Tong Y.-G.,State Key Laboratory of Pathogen and Biosecurity | Shi W.-F.,Institute of Pathogen Biology | Liu D.,CAS Institute of Microbiology | Qian J.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | And 53 more authors.
Nature | Year: 2015

A novel Ebola virus (EBOV) first identified in March 2014 has infected more than 25,000 people in West Africa, resulting in more than 10,000 deaths. Preliminary analyses of genome sequences of 81 EBOV collected from March to June 2014 from Guinea and Sierra Leone suggest that the 2014 EBOV originated from an independent transmission event from its natural reservoir followed by sustained human-to-human infections. It has been reported that the EBOV genome variation might have an effect on the efficacy of sequence-based virus detection and candidate therapeutics. However, only limited viral information has been available since July 2014, when the outbreak entered a rapid growth phase. Here we describe 175 full-length EBOV genome sequences from five severely stricken districts in Sierra Leone from 28 September to 11 November 2014. We found that the 2014 EBOV has become more phylogenetically and genetically diverse from July to November 2014, characterized by the emergence of multiple novel lineages. The substitution rate for the 2014 EBOV was estimated to be 1.23 × 10 â '3 substitutions per site per year (95% highest posterior density interval, 1.04 × 10 â '3 to 1.41 × 10 â '3 substitutions per site per year), approximating to that observed between previous EBOV outbreaks. The sharp increase in genetic diversity of the 2014 EBOV warrants extensive EBOV surveillance in Sierra Leone, Guinea and Liberia to better understand the viral evolution and transmission dynamics of the ongoing outbreak. These data will facilitate the international efforts to develop vaccines and therapeutics. © 2015 Macmillan Publishers Limited. All rights reserved.


Lu H.-J.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Qian J.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Kargbo D.,Ministry of Health and Sanitation | Zhang X.-G.,Centers for Disease Control and Prevention | And 18 more authors.
Emerging Infectious Diseases | Year: 2015

During 2014–2015, an outbreak of Ebola virus disease (EVD) swept across parts of West Africa. The China Mobile Laboratory Testing Team was dispatched to support response efforts, during September 28–November 11, 2014, they conducted PCR testing on samples from 1,635 suspected EVD patients. Of those patients, 50.4% were positive, of whom 84.6% lived within a 3-km zone along main roads connecting rural towns and densely populated cities. The median time from symptom onset to testing was 5 days. At testing, 75.7% of the confirmed patients had fever, and 94.1% reported at least 1 gastrointestinal symptom, all symptoms, except rash and hemorrhage, were more frequent in confirmed than nonconfirmed patients. Virus loads were significantly higher in EVD patients with fever, diarrhea, fatigue, or headache. The case-fatality rate was lower among patients 15–44 years of age and with virus loads of <100,000 RNA copies/mL. These findings are key for optimizing EVD control and treatment measures. © 2015, Centers for Disease Control and Prevention (CDC). All rights reserved.


Zhang K.,Peking Union Medical College | Zhang K.,Academy of Military Medical Science of PLA | Zhang Z.,Academy of Military Medical Science of PLA | Zhang Z.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | And 22 more authors.
Virus Research | Year: 2013

Replication and transmission of avian influenza virus (AIV) in domestic dogs and cats may pose a risk to humans. The susceptibility of cats and dogs to H9N2 influenza virus was evaluated by intranasally or orally inoculating animals with an H9N2 influenza virus. Cats had recoverable virus in respiratory tissues and the olfactory bulb three days post-inoculation and shed H9N2 virus into nasal washes and pharyngeal swabs from day 2 through day 10 post-inoculation. Virus was recovered from respiratory tissues of dogs three days post-inoculation, but was not detected in nasal washes or pharyngeal swabs. While no virus shedding or replication was detected in cats or dogs following consumption of H9N2-infected chicks, one of two cats and one of two dogs seroconverted. Two of three naïve contact cats seroconverted following co-housing with cats that were intranasally inoculated with H9N2 virus, whereas none of the three naïve contact dogs seroconverted. Our results demonstrate that H9N2 AIV can infect domestic cats and dogs via the upper respiratory tract and indicate that cats are more susceptible than dogs to H9N2 AIV. These findings suggest that domestic dogs and cats may serve as host species contributing to the adaptation of H9N2 viruses in mammals. © 2013 Elsevier B.V.


Zhang M.,Jilin University | Zhang M.,Academy of Military Medical science of PLA | Wang J.,Jilin University | Wang J.,Academy of Military Medical science of PLA | And 15 more authors.
International Journal of Oncology | Year: 2013

Apoptin is a chicken anemia virus-derived, p53-independent, bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis in various human tumor cells, but not in normal cells. To explore the use of apoptin in tumor gene therapy, we assessed a recombinant adenovirus expressing the apoptin protein (Ad-hTERTp-E1a-Apoptin) in order to determine its lethal and growth-inhibitory effects on PC-3 and RM-1 cells in vitro and its antitumor effect on solid tumors in vivo. 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide (MTT), acridine orange (AO)/ethidium bromide (EB), 4'-6-diamidino-2-phenylindole (DAPI), and Annexin V assays showed that Ad-hTERTp-E1a-Apoptin inhibited the proliferation of PC-3 and RM-1 cells in vitro by inducing apoptosis of prostate cancer cells, and that this inhibitory effect was dose and time-dependent. In the animal models, Ad-hTERTp-E1a- Apoptin significantly inhibited tumor growth and extended the lifespan of animals. Experimental results indicate that Ad-hTERTp-E1a-Apoptin has a potential application in tumor gene therapy.


Wang L.,Academy of Military Medical Science of PLA | Wang L.,Jilin Agricultural University | Wang L.,Key Laboratory of Jilin Province for Zoonosis Prevention and Control | Li T.,Academy of Military Medical Science of PLA | And 24 more authors.
Acta Theriologica Sinica | Year: 2012

In this study, the CDV originated from lesser pandas was isolated and served as the template for the infectious cDNA clone of CDV. After genome sequencing, seven cDNA fragments containing full-length CDV genome sequence were obtained by RT-PCR. The seven different fragments were digested and spliced together, and then inserted into eukaryotic expression vector pCI. Thus, we obtained the full-length cDNA of CDV lesser panda named pCI-CDV-LP. Furthermore, three helper plasmids were constructed by cloning the N, P, L protein ORF of lesser panda strain CDV in pCI vectors, respectively. The sequences of nuclease and CDV cDNA in pCI vector were verified by restriction enzyme digestion and sequencing. The full-length plasmid and three helper plasmids were then co-transfected into BSR cells using transfection reagent Lipofectamine ™2000. The results of RT-PCR, indirect immunofluorescence assay and viral infection assay showed that the lesser panda CDV was rescued successfully, and the reverse genetics of lesser panda CDV was built successfully. The results of the current investigation provide an important insight into the pathogenesis of canine distemper virus.

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