1.ISOLATION OF FEMALE- SPECIFIC SEQUENCE OF SCHISTOSOMA JAPONICUM
Chinese Journal of Zoonoses 2000;(2):72-73
Aim To isolate the female-specific sequence of Schistosomajaponicum S. J. in order to provide a basis for researching vaccines of anti-fecundity. Methods Representational difference analysis (RDA) was used to obtain a female-specific sequence of S.j. by the male S.j. genomic DNA substractive hybridization with the female. The fragment was about 562bp. Results This sequence was labeled as Dig probe by PCR technique to hybridize with the restriction fragments of each sex and the tester as well as the driver on the nylon membranes.The antiboly of Dig、 NBT、 and X-phosphate solution were utilized to immunoassay the result of hybridization. The result showed that the probe can hybridize to the tester but can not hybridize to the driver. Conclusion The result suggested that the fragment of approx. 562bp was the female-specific sequence of Schistosoma japonicum.
2.Epidemiological Investigation of Angiostrongylus cantonensis in Jiangmen of Guangdong Province
Yun ZHANG ; Di HUANG ; Qiming TAN ; Daixiong CHEN ; Ximei ZHAN
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective To make an epidemiological survey on Angiostrongylus cantonensis in Jiangmen City of Guangdong Province. Methods From October 2006 to November 2007, the characteristics of A. cantonensis infection were investigated in Jiangmen district in various hosts, including the third stage larva infection in the snails Achatina fulica and Pomocea canaliculata by digestion method, and the adult A. cantonensis in rats by the dissection of heart and lungs. Relevant symptoms and dietary habits in Jiangmen residents who were randomly recruited were also investigated by questionnaire, and the specific IgG and IgM antibodies against A. cantonensis in their sera were detected by ELISA. Results 695 A. fulica and 720 P. canaliculata were examined. The infection rate of third stage larva of A. cantonensis were 45.0% and 1.8% respectively, with an infectiosity of 53.74?147.30 and 5.23?8.51 respectively. Natural infection rate of A. cantonensis in all 229 rats was 4.4%. Among the 300 people surveyed, 11.3% had a history of eating raw or undercooked fish and shrimp, 5.3% directly or indirectly exposed to A. fulica or P. canaliculata. The positive rate of specific IgG antibody against A.cantonensis for serum samples among residents was 14.0% (42/300), and 5 serum samples in the 42 positive samples showed specific IgM antibody, with a positive rate of 1.7%. Conclusion Jiangmen district is an endemic area of A. cantonensis, and the local residents are under the risk of infection.
3.KPC carbapenemases among Enterobacteriaceae with carbapenem non-susceptibility
Hui HUANG ; Shumin SHE ; Ximei ZHAN ; Duorong WU
Chinese Journal of Zoonoses 2015;(3):247-250
To investigate the prevalence and gene types of KPC in Enterobacteriaceae strains isolated from 4 tertiary general hospitals in Hainan area ,a total 43 isolates which were resistant or intermediate to imipenem or ertapenem were collected from sterile sites between August 2012 and June 2013 from 4 tertiary general hospitals in Hainan area .Modified Hodge Tests (M HT ) were performed for KPC phenotype screening .PCR amplification and DNA sequence were performed to analyze the encoding genes of KPC .Results showed that in the 43 isolates ,21 strains were positive in M HT .PCR and DNA sequence analysis confirmed that 3 isolates produced KPC‐2 .It's suggested that there were the Enterobacteriaceae carrying KPC in Hain‐an area .The encoding genes were KPC‐2 .The KPC gene could be horizontally transmitted by plasmid among different groups of bacteria .It is important to control the transmission of these Enterobacteriaceae carrying KPC .
4.Dynamic change of parasites in the brains of BALB/c mice infected with Angiostrongylus cantonensis
Yunhu TU ; Conghua LI ; Weihong XU ; Zheng ZHANG ; Dajian LONG ; Pan YUE ; Zhenyu QU ; Ximei ZHAN
Chinese Journal of Zoonoses 2009;(12):1192-1194
To observe the dynamic change of parasites in the brains of BALB/c mice infected with Angiostrongylus cantonensis in order to explore its possible mechanism of pathogenesis', BALB/c mice infected with the III stage larvae of A.cantonensis were observed and killed in different times after infection. The number and distribution of parasites in the brains of the infected mice were observed microscopically and macroscopically. It was found that the larvae of A.cantonensis were distributed in the cerebrum and cerebellum of mice in accordance with the rule of parasitization of worms in the host, i.e.multiplication at first and then dropping in number. And the places where the parasites located were damaged due to mechanical action or inflammatory reactions. The time of onset of symptoms, such as ataxia and twitch was coincided with the dynamic changes in the brains of the infected mice.
5.Identification of interaction site of MIC2 with aldolase in Toxoplasma gondii
Bin ZHENG ; Zhikui YIN ; Zhijun YAO ; Haizhu ZHANG ; Hongbin REN ; Ximei ZHAN
Chinese Journal of Zoonoses 2014;(7):698-700,708
In this study ,we aim to identify the protein interaction site of microneme protein 2 (MIC2) and aldolase in Toxoplasma gondii .The tryptophan (Trp ,W) at site 767 of carboxyl terminus of MIC2 (MIC2C) was mutated into alanine (Ala ,A) by site-directed mutagenesis to construct plasmid MIC2C W/A/pGEX-4T-1 .The mutant protein GST-MIC2C W/A was expressed in E .coli upon IPTG induction .Glutathione sepharose beads were incubated with GST-MIC2C W/A and GST-MIC2C respectively ,then incubated with tachyzoite lysates ,and bound proteins were eluted using sample buffer .Eluants were resolved by SDS-PAGE and Western blot .A protein band specifically recognized by anti-aldolase antibody was detected in prod-ucts coming from GST pull-down of GST-MIC2C ,but not in pull-down products coming from GST-MIC2C W/A .With muta-tion of MIC2C W767 to A ,MIC2 protein lost the binding ability to aldolase .Tryptophan (W767 ) was the protein interaction site of MIC2 and aldolase in T .gondii .
6.Octreotide inhibits proliferation and extracellular matrix synthesis in hepatic stellate cells
Zhirong WANG ; Ximei CHEN ; Dingguo LI ; Xin HUANG ; Hongshan WEI ; Yutao ZHAN ; Yuqin WANG ; Hanmin LU
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To investigate the effects of octreotide (Oct) on the proliferation and extracellular matrix (ECM) synthesis in hepatic stellate cells (HSCs). METHODS: HSCs were isolated from normal male Sprague-Dawley rat liver by a combination of pronase-collagenase perfusion and density gradient centrifugation. The concentration of 2.5 ?g/L transforming growth factor ?1 (TGF?1) was used in all the experiment settings. Oct at concentrations of 0.01 mg/L ,0.1 mg/L,1 mg/L and 10 mg/L,respectively,or 0.01 mg/L Oct + TGF?1,0.1 mg/L Oct+TGF?1,1 mg/L Oct+TGF?1,10 mg/L Oct+TGF?1 were respectively added to the cultured HSCs. Effects of Oct on HSC proliferation and ECM synthesis were respectively determined by MTT method,-TdR and -proline incorporation,or radioimmunoassay. RESULTS: Oct inhibited MTT intake by cultured hepatic stellate cells and down-regulated -TdR incorporation,compared with control group. The concentrations of hyaluronic acid,laminin,collagen type IV in the culture supernatant and -proline incorporation in HSCs were decreased by Oct. TGF?1 obviously up-regulated proliferation and ECM synthesis in cultured HSCs,and Oct significantly blocked these actions. CONCLUSION: Oct inhibited proliferation and ECM synthesis in cultured HSCs,and elicited the effects of anti-hepatofibrogenesis.
7.Inhibiting severe acute respiratory syndrome-associated coronavirus by small interfering RNA.
Renli ZHANG ; Zhongmin GUO ; Jiahai LU ; Jinxiu MENG ; Canquan ZHOU ; Ximei ZHAN ; Bing HUANG ; Xinbing YU ; Min HUANG ; Xinghua PAN ; Wenhua LING ; Xigu CHEN ; Zhuoyue WAN ; Huanying ZHENG ; Xinge YAN ; Yifei WANG ; Yanchao RAN ; Xinjian LIU ; Junxin MA ; Chengyu WANG ; Biliang ZHANG
Chinese Medical Journal 2003;116(8):1262-1264
OBJECTIVETo evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases.
METHODSVero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells.
RESULTSFive pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect.
CONCLUSIONsiRNA may be effective in inhibiting SARS-associated coronavirus replication.
Animals ; Cercopithecus aethiops ; RNA, Small Interfering ; pharmacology ; SARS Virus ; drug effects ; Transfection ; Vero Cells ; Virus Replication ; drug effects