1.Research progress on Salmonella Derby
Huijuan ZHENG ; Zhiming PAN ; Xinan JIAO
Chinese Journal of Zoonoses 2017;33(7):642-645
Salmonella Derby is recognized as a major human food-borne pathogen causing food poisoning,septicaemia and other symptoms.Meanwhile,it can represent a severe threat to livestock breeding and health.The objective of this review is to summarize novel research progress on epidemic,genomics,pathogenic mechanism of Salmonella Derby for provide reference to related research.
2.Development of swine Salmonella vaccine
Shanshan ZHU ; Yingying ZHOU ; Shizhong GEN ; Zhiming PAN ; Xinan JIAO
Chinese Journal of Zoonoses 2017;33(6):548-552
Salmonella is an important zoonosis.China is the largest pork consumer.Contaminated pork is the main source of Salmonella disease at home and abroad.There are many swine Salmonella vaccine in use in Europe.Now the current swine Salmonella vaccine is a live attenuated vaccine,Salmonella can reduce colonization in pigs,and can induce immune response efficiently after the two immunizations.The control effect is poor,at this stage of swine vaccine cross protection research,type of Salmonella and vaccine antigen are not the same.This paper reviews the research progress of swine Salmonella vaccine.
3.Research progress on T3SS relative effector protein of Salmonella
Peipei TANG ; Zhijie LIN ; Zhiming PAN ; Xinan JIAO
Chinese Journal of Zoonoses 2015;(3):277-280,288
Salmonella is a pathogenic bacteria to human and animals ,which can cause seriously complication and death . Salmonella pathogenicity is from the reactions of SP‐1 and SP‐2 T3SS effector proteins .In this paper ,the functions of different T3ss effector proteins in different infection periods is reviewed to provide a reference for further understanding the pathogenesis mechanisms of Salmonella .
4.The analysis of quinolone resistance of the avian Escherichia coli and Salmonella isolates from 1993 to 2008
Weijuan PAN ; Xiang CHEN ; Xiaoquan WANG ; Qiuxia CONG ; Zhiming PAN ; Song GAO ; Xinan JIAO
Chinese Journal of Zoonoses 2009;(7):630-635
Quinolones are broad-spectrum antibacterial agents used in human and veterinary medicine, and their extensive use have been associated with a rise of the quinolone resistance. In the present study, the quinolone resistance of avian E.coli and Salmonella isolates was evaluated and compared, in which 344 avian E.coli and 224 Salmonella isolates from 1990s were serogrouped with antisera and thc antimicrobial susceptibility test to 10 quinolones was carried out by using the Kirby-Bauer method recommended by Clinical and Laboratory Standards Institute (CLSI). It was demonstrated that the 344 isolates of avian E.coli distributed in 27 serogroups and 68.90% (237/344) of the isolates belonged to four O-serogroups: i.e. O1, O2, O18, O78, and the 224 isolates of avian Salmonella were all determined to be Salmonella pullorum. The drug-resistance rate of avian E. coli isolates to nalicixic acid from 1993-1999 was more than 60%(64.43%,131/181), whereas those of isolates to 9 antibiotics from 2000-2008 had a drug-resistance rates of more than 60%, namely,nalicixic acid(92.02%), fleroxacin(79.75%), pipemidic acid(79.14%), enrofloxacin(78.53%), enoxacin(76.07%), lomenfloxacin(74.85%), ciprofloxacin(69.33%), norfloxacin(63.80%) and ofloxacin(61.35%). For the 4 O-serogroups of the avian E.coli isolates, the drug-resistance rates of more than 50% to antimicrobials were as follows: O78 isolates to 7 antimicrobials;O18 isolates to 5 antimicrobials, and O1 and O2 isolates just to 3 antimicrobials. The quinolone resistance of Salmonella isolates was much lower than E.coli, in which 101 salmonella isolates from 1993-1999 were all susceptible to quinolones. Nalicixic acid resistance of salmonella isolate firstly appeared in 2000, and the drug-resistance rate of salmonella isolates from 2000-2008 was found to be more than 60% for nalicixic acid(83.74%), but those to other quinolones were comparatively lower. These results indicated that the quinolone resistance of avian E.coli and salmonella were increasing in the past two decads because of the over-use of antibiotics.
5.Immune responses specific for ESAT-6 expressed by recombinant Salmonella typhimurium
Hui ZHANG ; Liu LIU ; Shizhong GENG ; Maozhi HU ; Ke WEN ; Zhiming PAN ; Xinan JIAO
Chinese Journal of Microbiology and Immunology 2009;29(6):528-532
Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.
6.Progress of Listeria monocy togenes as tumor vaccine vector
Feifei DUAN ; Yuelan YIN ; Meiqin KANG ; Weijun TAN ; Chengwu TAO ; Zhiming PAN ; Jinlin HUANG ; Xinan JIAO
Chinese Journal of Zoonoses 2014;(7):743-746,752
Listeriamonocytogenes is a facultative intracellular bacterium that enters professional antigen presenting cells , presents passenger antigens to the major histocompatibility complex class I and II pathways ,then elicits CD+4 and CD+8 T-cell-mediated immune responses .It was demonstrated that attenuated Listeriamonocytogenes as a novel live vaccine vector in deliv-ering tumor antigens of cervical cancer and melanoma etc .,could induce strong protective immune response ,and shows effec-tive antitumor immunotherapeutics .This review discussed the characteristics of immune responses elicited by Listeria monocy-togenes ,and the progress of its antitumor immunotherapeutics as delivery vaccine vector .
7.Study on CD8~+ T cell epitope expressed by eukaryotic plasmid delivered by attenuated E.coli
Zhiming PAN ; Xiaoming ZHANG ; Xinan JIAO ; Loman RICHARD ; Leclerc CLAUDE ; Xiufan LIU
Chinese Journal of Immunology 1985;0(03):-
Objective:To analyze the efficiency of delivery for CD8+ T cell epitope by attenuated E.coli vector.Methods:The recombinant E.coli strain 13A(pG2F),harbouring the eukaryotic expression plasmid pG2F with CD8+ T cell epitope of Ovalbumin (OVA) and green fluorescent protein (GFP) marker at the C-terminal was used to infect into LKb cells,bone marrow dendritic cells (BMDC).The efficiency of presentation for CD8+T cell epitope delivered by recombinant bacteria was analyzed by in vitro antigen presentation assay.C57BL/6 mice were immunized intravenously with 13A(pG2F).Murine IFN-? secreting cells were detected in murine splenocytes by enzyme-linked immunospot assay (ELISPOT).Results:After the infection of LKb cells,BMDC by recombinant bacteria,about 0.3%~4% of cells were GFP positive.The results indicated that attenuated strain 13A could deliver the eukaryotic expression plasmid into mammalian cells.At 2 h post infection,CD8+ T cell epitope was presented on the surface of those LKb,BMDC cells infected by 13A (pG2F) could be recognized by B3Z T hybridoma cells.The presentation efficiency of LKb cells for OVA CD8+ T cell epitope was increased at 48h after infection.Furthermore,the presentation efficiency of BMDC was higher than those of LKb cells under the same condition.The recombinant bacteria 13A(pG2F) could induce cellular immune responses in C57BL/6 mice.Conclusion:Attenuated E.coli can effectively deliver the CD8+ T cell epitope in vitro and in vivo.
8.Epidemiology of listeriosis infection in humans and animals, China, 2002-2012
Kai LIAN ; Weijun TAN ; Dan ZHAO ; Yuelan YIN ; Guoliang WANG ; Feifei DUAN ; Zhiming PAN ; Jinlin HUANG ; Xinan JIAO
Chinese Journal of Zoonoses 2014;(10):1033-1038
The prevalence of human and animal listeriosis for nearly 11 years in China was investigated in this study . The literature information about listeriosis in China from 2002 to 2012 was collected through retrieval system to make clinical and epidemiological statistical analysis of listeriosis .Cases of listeriosis were reported in 27 (79% ) provinces of China .The re-sult showed that animal listeriosis was reported for 123 times ,among these reports ,most were from pigs (39% ) ,and the sheep was in second place .Central nervous system infection was the main clinical manifestation of listeriosis in animals (72% ) . For human listeriosis ,84 clinical cases of listeriosis were reported ,including 35% cases in non-perinatal stage and 65% cases in perinatal stage .The main clinical manifestation of listeriosis was septicemia (51% ) .According to the result of investigation about listeriosis based on literatures information ,Listeriamonocytogenes caused humans and animals listeriosis annually ,which were reported in most provinces of China .The epidemic characteristics for listeriosis suggested that it was essential to strength-en the prevention and control of listeriosis .
9.Expression and antiviral assay of bovine interferon-gamma.
Zhengzhong 'U ; Xiang CHEN ; Fengli SHAN ; Chuang MENG ; Lin SUN ; Jinlin HUANG ; Zhiming PAN ; Shizhong GENG ; Xinan JIAO
Chinese Journal of Biotechnology 2011;27(2):269-276
Bovine interferon-gamma (BoIFN-gamma) gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from total RNA of bovine spleen lymphocytes stimulated with ConA. The products of RT-PCR were cloned into pVAX1 vector, positive recombinant clone was identified by restriction enzyme digestion and sequencing. The recombinant plasmid pVAX1-BolFN-gamma was transfected into COS-7 cells mediated by lipofectine, indirect immunofluorescent assay analysis confirmed that rBoIFN-gamma was expressed in COS-7 cells. BoIFN-gamma gene (without signal peptide) was cloned into pET-30a(+) and pGEX-6p-1 vector, and transformed into the Escherichia coli cells. After optimizing the induction condition, SDS-PAGE analysis showed that the expression products were all found in soluble form and had a molecular weight of 23 kDa and 43 kDa respectively. BoLFN-gamma precursor gene (with signal peptide) was cloned into transfer vector pFastBac 1, and transformated into DH10Bac E. coli cells. By site-specific transposition, BoIFN-gamma gene was integrated into shuttle vector Bacmid, and transfected into the Sf9 insect cells mediated by lipofectine to produce recombinant baculovirus. Indirect immunofluorescent assay analysis confirmed that rBac-BoLFN-gamma was expressed successfully in Baculovirus vector system. The antiviral activities of rHis-BoIFN-gamma, rGST-BoIFN-gamma and rBac-BoIFN-gamma were up to 8.389 x10(7) U/mg, 6.554 x10(5) U/mg and 4.096 x 10(4) U/mL respectively, which were analyzed in MDBK/VSV system. A sandwich ELISA was established using monoclonal antibodies 3E6 and 5G4, which can detect BoIFN-gamma in quantity and provide a useful method for the clinical practice and research of BolFN-gamma.
Animals
;
Antiviral Agents
;
pharmacology
;
Baculoviridae
;
genetics
;
metabolism
;
COS Cells
;
Cattle
;
Cercopithecus aethiops
;
Escherichia coli
;
genetics
;
metabolism
;
Genetic Vectors
;
genetics
;
Interferon-gamma
;
biosynthesis
;
genetics
;
pharmacology
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
pharmacology
;
Transfection
10.Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155
Wanli QIN ; Yujie XU ; Zhenzhen PAN ; Xiaohui LI ; Zhenhua WANG ; Jianping SONG ; Qin XU ; Xinan HUANG ; Changqing LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(14):29-35
ObjectiveTo observe the effects of Fuzitang (FZT) on the proliferation of MH7A cells, the human rheumatoid arthritis synovial fibroblasts, and the expression of miR-155 and explore its anti-rheumatoid arthritis mechanism. MethodMH7A cells were cultured in vitro and divided into a blank group, high- (25 g·L-1) and low-dose (12.5 g·L-1) FZT groups, and a positive drug group (hydroxychloroquine, 0.006 25 g·L-1). The cell proliferation was detected by cell counting kit-8(CCK-8) method, and the change in the MH7A cell cycle was detected by flow cytometry. The mRNA expression of miR-155 and its downstream genes, including SH2 domain-containing inositol 5-phosphatase-1(SHIP-1), protein kinase B 3(Akt3), and mammalian target of rapamycin(mTOR), was detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the protein expression of phosphatidylinositol 3-kinase (PI3K), Akt3, and mTOR was detected by Western blot. ResultFZT in vitro in a concentration of 6.25 g·L-1 above could inhibit the proliferation of MH7A cells in the significant dose- and time-effect manner. Compared with the blank group, the FZT groups showed increased proportions of cells in the G2/M phase (P<0.05), and the high-dose FZT group showed a decreased proportion of cells in the G0/G1 phase (P<0.05). The arresting effect of FZT on the cell cycle was in a significant dose-effect manner. Compared with the blank group, the FZT groups showed down-regulated miR-155 and mTOR mRNA expression (P<0.05), and the high-dose FZT group showed up-regulated SHIP1 mRNA expression and down-regulated Akt3 mRNA expression (P<0.05). Compared with the blank group, the FZT groups showed reduced protein expression of PI3K, Akt3, and mTOR (P<0.05). ConclusionFZT can significantly inhibit the proliferation of MH7A cells, and the mechanism is related to the promotion of the expression of SHIP-1 and down-regulation of the gene expression of the PI3K/Akt3/mTOR signaling pathway by down-regulating the expression of miR-155.