Table of Content

30 August 2000, Volume 18 Issue 4

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论著

INDUCIBLE EXPRESSION OF MSP1 GENE OF PLASMODIUM FALCIPARUM BY A TETRACYCLINE-CONTROLLED PROMOTER

QIANFeng;PANWei-qing

2000, 18(4):  1-196. 

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　Objective] To express the entire MSP1 gene of Plasmodium falciparum and its C-terminal 42 kDa fragment using a tetracycline-controlled P LtetO-1 promoter. [Methods] The entire MSP1 gene and 42 kDa fragment gene were cloned into the plasmid of pZE11,and transformed into E coli DH5αZ1. Restriction e nzyme analysis, SDS-PAGE and Western blotting were used to examine two recombinant plasmids and their expression in E coli DH5αZ1. [Results ] The recombinant plasmids of pZE11/MSP1 and pZE11/MSP1-42 were constructed successfully. The expressive products about 190 kDa and 42 kDa of two genes in E coli DH5αZ1 were identified by SDS-PAGE and Western blotting. [Conclusion] Tightly controlling expression of the MSP1 gene in E coli is essential to reduce the toxicity of the product to its host cells as well as to provide a feasibility to construct Salmonella vaccine strain which can inducibly express the important malarial vaccine candidate gene.


EFFECT OF ANTI-MOSQUITO-MIDGUT ANTIBODIES ON THE DEVELOPMENT OF OOCYSTS OF PLASMODIUM YOELII IN ANOPHELES STEPHENSI

WEIQiu-fen;GAOXing-zheng

2000, 18(4):  2-199. 

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　Objective] To study the effect of anti-mosquito-midgut antibodies on the development of P.yoelii in A.stephensi . [Methods] Ookinetes and oocysts in midgut and sporozoites in the salivary glands of mosquitoes were examined respectively at 14 h and 9 d and 12 d after the mosquitoes ingested infected blood containing anti-midgut antiboides by membrane feeders. The effect of anti-midgut antibody level and repeated feedings on the development of the oocysts in the mosquitoes was also observed. [Results] The anti-midgut antibodies reduced evidently not only the infection rates of the oocysts and the number of the sporozoites in salivary gland but also infection degree of ookinetes and the index of oocysts in the mosquitoes compared with control group ( P <0 05). The higher the antibody level, the more the times for mosqitoes digested antibodies, and the fewer the number of oocysts. [Conclusion] The anti-mosquito-mindgut antibodies inhibited ookinetes and oocysts of P.yoelii in A.stephensi , with oocysts in particular.


PLASMODIUM FALCIPARUM : ANTIMALARIAL ACTIVITIES OF ANTISENSE OLIGODEOXYRIBONUCLEOTIDES TARGETED TO HISTIDINE-RICH PROTEIN GENES IN VITRO

YUXin-bing;FANGJian-min;LUOShu-hong

2000, 18(4):  3-203. 

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　Objective] To study the inhibition of growth of Plasmodium falciparum cultured in vitro with antisense oligodeoxyribonucleotide (AS ODN) against histidine-rich proteins (HRP). [Methods] The ODNs against HRP Ⅱ and HRP Ⅲ were synthesized and used to study the antimalarial activities in vitro . Plasmodium falciparum (FCC1/HN strain, China) were exposed to AS DONs for 48 h, and the growth inhibi tion was determined by microscopic examination. [Results] At 1 μmol/L, all ODNs inhibited parasite growth and development in a target-indepentent manner. However, when the ODN concentrations were between 0.01 and 0 5 μmol/L, the AS ODN significantly inhibited the growth and development of P falciparum compared with ODN controls ( P <0 01). Inhibition by the sense strand ODN did not differ significantly from the control group ( P> 0 05). [Conclusion] Blockade of the expression of HRPⅡ and HRPⅢ AS ODN could inhibit P falciparum cultured in vitro .


THE SCHIZONTOCIDAL ACTIVITY OF DAPHNETIN AGAINST MALARIA PARASITES IN VITRO AND IN VIVO

WANGQin-mei;NIYi-chang;XUYue-qin;HAShu-hua;CAIYue

2000, 18(4):  4-206. 

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　[Objective] To investigate the in vitro and in vivo schizontocidal activity of daphnetin. [Methods] Schizontocidal activity of daphnetin was tested using an in vitro assay based on the routine in vitro cultivation of P. falciparum FCC 1 strain. The in vivo antimalarial effects of daphnetin at various dosages were assessed in mice infected with P b.erghei ANKA according to “4-day suppress assay”. [Results] In vitro ,daphnetin exhibited potent schizontocidal activity comparable to chloroquine(CQ) at the dose range of 1～10 μmol/L. In vivo ,50 or 100 mg/kg.d -1 ×4 d daphnetin i.g. and 10,50 or 100 mg/kg.d -1 ×4 d dephnetin i.p. showed antimalarial efficacy comparable to CQ 10 mg/kg.d -1 ×4 d i.g. in mice infected with P. berghei ANKA, evaluated by both the reduction rate of parasitemia on D 4 and the average surviving days in 30 days. [Conclusion]Daphnetin displays certain schizontocidal activity both in vitro and in vivo .


THE ROLE OF CD8 + T CELLS IN THE PATHOGENESIS OF PNEUMOCYSTIS CARINII PNEUMONIA IN MICE DEPLETED OF CD4 + T CELLS

ANChun-li;SUXiao-ping;A.G.Harmsen

2000, 18(4):  5-212. 

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　Objective] To define the cell populations contributing to inflammation-associated respiratory impairment in Pneumocystis carinii pneumonia (PCP). [Methods] The host inflammation response was observed in CD4 +T cell-depleted mice and in CD4 + and CD8 + T cell-depleted mice infected with P carinii via intratracheal inoculation. [Results] Mice depleted of both CD4 + and CD8 + T cells developed infection with neither increased respiratory rate nor lung injury. In contrast, mice depleted of CD4 + T cells alone exhibited severe pulmonary inflammation, and increased respiratory rates. Respiratory compromise was associated with the presence of activated CD8 + cells and neutrophils in the BALF. [Conclusion] The host′s inflammatory cell response to P. carinii directly impairs pulmonary function and contributes to the pathogenesis of PCP, CD8 + T cells appear to play a major role.


GENE SEQUENCING FOR IDENTIFICATION OF PARAGONIMUS EGGS FROM A HUMAN CASE

CHANGZhen-shan;WUBuo;BlairD;ZHANGYong-nian;HULing;CHENShao-hong;CHENMing-gang;FENGZhen;GeorgeM.Davis

2000, 18(4):  6-215. 

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　Objective] To identifiy the etiologic agent from a paragonimiasis patient using molecular techniques. [Methosd] The complete nuclear ribosomal DNA second internal transcribed spacer (ITS2) gene sequence of eggs in sputum from a paragonimiasis patient was obtained by directly auto-sequencing its PCR product . ITS2 genes from eggs of Paragonimus westermani and Pagumogonimus skrjabini (both from animal hosts) were also sequenced for comparison. In addition , morphological comparisons were made with the eggs of the two species. [Results] The ITS2 gene from the human case was 100% identical with the sequence from the eggs of P westermani from an experimentally infected dog but only 92% identical with the sequence from the eggs of P skrjabini. Morphologically, the eggs from the human case more resembled those from P westermani infected dog. [Conclunsion] The patient was diagnosd to be suffered from paragonimiasis westermani by gene sequence analysis.


LOCALIZATION AND CHARACTERIZATION OF PARTIAL IMMUNODOMINANT ANTIGEN EPITOPES OF TRICHINELLA SPIRALIS

ZHANYan-ai;YANZi-zhu;WANWen;LUZai-ying;ZHUZhen-qin

2000, 18(4):  7-219. 

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　Objective] To screen and characterize immunodominant antigen epitopes on the soluble antigens of Trichinella spiralis (T s ) . [Methods]15 monoclonal antibodies (McAbs) against T s muscle larva(ML) soluble antigens were obtained by using hybridoma technique. The reactivity of monoclonal and polyclonal antibodies were tested by ELISA, Western blotting and indirect immunofluorescence assay(IFA). [Results] The Western blotting result showed that of the 15 McAbs, 6 could bind to the T s ML antigens displaying molecular weights of 40～70 kDa. Polyclonal sera could react with more than 10 bands having molecular weights of 20～200 kDa. Among the 6 McAbs, 4 could recognize epitopes on the cuticle surface and the other two could recognize epitopes on both the cuticle surface and the stichosome. [Conclusion]The antigen epitopes of T s recognized by 6 McAbs had been characterized.


GENE CLONING AND CHARACTERIZATION OF MITOCHONDRIA-RELATED PROTEIN OF SCHISTOSOMA JAPONICUM 

HUXue-mei**;WUHai-wei;ZHANGZhao-song;SUChuan;ZHAOwei;SHENLei;WANGRong-zhi;MALei;ZHOUJi-li;CHENShu-zhen;WUGuan-ling

2000, 18(4):  8-223. 

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　Objective] To subclone and characterize a cDNA clone coding for Schistosoma japonicum (S.j.) mitochondria-related protein. [Methods] The open reading frame of the fragment(Sj338/24) obtained from an adult worm cDNA library of S j. was analysed, at the upstream and downstream of the open reading frame(ORF) the primers A and B were designed,respectively, and the cDNA fragment was used as PCR template. The Sj338 gene fragment obtained was amplified by PCR method and then subcloned into pGEM-T vector for sequencing. The gene sequence was analyzed and the target fragment was restrictedly digested and subcloned into expression vector pGEX-6P-1. The expressed recombinant protein was purified and characterized. [Results] The cloned Sj338 gene was demonstrated to be 487 bp long containing one 459 bp ORF, encoding a protein with a molecular weight of 17 kDa.The nucleotide sequence of the cloned gene Sj338 had higher homology with those genes coding for mitochondrial outer membrane protein of Homo sapiens and Rattus norvegicus . The recombinant construct of pGEX-6P-1/Sj338 could be expressed efficiently and the antigenicity of its product rSj338 has been demonstrated by Western blotting. [Conclusion] Sj338 may be the gene coding for S j. mitochondria-related protein and the recombinant protein may be used as a new vaccine candidate .


INVESTIGATION OF ANTI- TOXOPLASMA GONDII ANTIBODIES IN IMMUNODEFICIENT PATIENTS

WANGBao-lin;PANXiao-zhang;YINYou-kuan;WENGXin-hua

2000, 18(4):  9-226. 

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　Objective] To investigate the presence of anti- Toxoplasma gondii antibodies in immunodeficient patients. [Methods] T gondii -specific immunoglobulin G (IgG) antibodies in serum samples from 371 immunodeficient patients were detected by enzyme-linked immunosorbent assay (ELISA). The patients were with solid malignancies (including untreated digestive system malignancies and solid malignancies received chemotherapy), chronic liver diseases, patients received immunosuppressant therapy (dermatomyositis, psoriasis, pemphigus, post-renal transplantation, systemic lupus erythematosus and other miscellanies), lymphoma, leukemia and diabetes. 100 normal serum samples served as controls. [Results] The positive rate of patients with solid malignancies received chemotherapy, solid malignancies received chemotherapy, chronic liver diseases, systemic lupus erythematosus and leukemia was 19 0%, 33 3%, 16 5%, 45 4% and 20 0%, respectively,being significantly higher than that of the control group( P <0 05). [Conclusion] The immunosuppressed patients are highly predisposing to secondary T.gondii infection .


COMPARISON OF CIRCULATING ANTIGEN DETECTION IN SCHISTOSOMIASIS JAPONICA PATIENTS USING TWO METHODS OF ELISA 

QIULi-zhuZHANGYong-hongLIHaoXUEHai-chou

2000, 18(4):  10-228. 

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　Objective] To compare 2 kinds of ELISA methods for detecting circulating antigen in schistosomiasis patients. [Methods] Monoclonal antibody-based dot-ELISA and sandwich ELISA. [Results and Conclusion] The positive rates of dot-ELISA were higher than or similar to those of sandwich ELISA,the former being 48 0%～98 8% and the latter being 12 0%～93 8%. The specificities were 95 4%～100% and 90 0%, respectively.


ESTABLISHMENT OF AN ANIMAL MODEL OF CAPILLARIA HEPATICA

YANGFa-zhu;HUANGXiao-hong;TUZhao-ping;ZHANGYing-zhen

2000, 18(4):  11-231. 

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　Objective] To establish an animal model of Capillaria hepatica. [Methode] Sixteen rats and two cats were infected with the embryo eggs of Capillaria hepatica through mouth. [Results] Of 16 infected rats, 2 were negative, 14 were positive with Capillaria hepatica . 2 cats were negative.[Conclusion] Rats could be used as an animal model of Capillaria hepatica .


防治经验

INVESTIGATION ON SCHISTOSOMA JAPONICUM INFECTION IN RODENTS IN YUNNAN PROVINCE

YANGGuang-rong;WUXing;XIONGMeng-tao;FANCongzheng;WUHesong;TAOKaihui

2000, 18(4):  12-235. 

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　To investigate the role of rodents in the transmission of schistosomiasis in endemic areas in Yunnan Province.[Methods]Kato-Katz method was used to study the infection rate of rodents,humans and domestic animals.[Results] The infection rate was 0 9%(32/3 411)for rodents,15 6%(461/2 964) for humans,and 9.6% (239/2 482),for domestic animals.The EPG() was 5 8 for rodents,1 8 for humans,0 1 for cattle,0 1 for horses and 0 02 for pigs.[Conclusion]The rodents played a minor role in the transmission of schistosomiasis in the plateau area of Yunnan province. [WT5”FZ]