Table of Content

30 August 1999, Volume 17 Issue 4

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简报

CLONING,SEQUENCING AND EXPRESSION OF THE FULL-LENGTH GENE ENCODING PARAMYOSIN OF SCHISTOSOMA JAPONICUM IN VIVO *

ZHOUShenghua;LIUShuxian;SONGGuangcheng;XUYuxin

1999, 17(4):  2-199. 

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　AIM: To clone and sequence the gene encoding paramyosin of S. japonicum (Chinese strain) and to study the expression of the DNA-based vaccine encoding the full-length paramyosin of S. japonicum in vivo . METHODS: Total RNA was isolated from adult S. japonicum using TRIzol reagent. The full-length cDNA encoding paramyosin of S. japonicum was amplified by RT-PCR and cloned into pGEM-T vector and sequenced by the method of dideoxy-mediated chain-termination. The cDNA encoding paramyosin of S. japonicum was subcloned into the expressive plasmid vector pCDNA/AMP(pCMV-Sjc97), and the recombinants were identified by restriction enzyme digestion and sequencing.The immunofluorescence assay was used to study the expression of Sjc97 in vivo in mice. RESULTS AND CONCLUSION: The 2.6 kb cDNA encoding the full-length paramyosin of Chinese S. japonicum has been successfully cloned and sequenced for the first time. The full-length sequence of paramyosin of S. japonicum was determined. Comparison of the nucleotide sequence and the deduced amino acid sequence of Sjc97 with that of S.japonicum paramyosin(Philippine strain)(Sjp97), S.japonicum paramyosin(Japanese strain)(Sjj97), S.mansoni (Sm97) , B6 and Y6 clone ( the partial cDNA encoding paramyosin of Chinese strain) showed that Sjc97 differed from Sjp97 by 16/2 601 nucleotide and 3/866 amino acid substitutions (99.4% on nt-levlel and 99.7% on aa-level in homology); from Sjj97 by 20/2 601 nucleotide and 2/866 amino acid substitutions (99.2% on nt-levlel and 99.8% on aa-level in homology); and from B6 by 11/1 329 nucleotide and 1/443 amino acid substitutions (99.0% on nt-level and 99.8% on aa-level in homology); from Y6 by 13/1 329 nucleotide and 1/443 amino acid substitutions (98.9% on nt-levlel and 99.8% on aa-level in homology); Sjc97 differed from the Sm97 by 2 235/2 601 nucleotide and 34/866 amino acid (91.0% on nt-levlel and 96.0% on aa-level in homology). The plasmid expression vector encoding the full-length paramyosin of Chinese S. japonicum has been successfully constructed. The pCMV-Sjc97 vaccine could express Sjc97 protein in vivo in mice after intramuscular immunization.


论著

HYBRIDIZATION EXPERIMENTS USING ANOPHELES MINIMUS FROM HAINAN AND YUNNAN

SHIHuanhuan;TIANChunlin;HEDengxian

1999, 17(4):  3-202. 

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　AIM: To observe whether there were any intra-species differences between Anopheles minimus from Hainan(H) and Yunnan(Y). METHODS: Anopheles minimus were collected from cattle shed on the spot. Each isofemale line was set up in the laboratory. Hybridization experiments were conducted by using forced mating between Anopheles minimus from Hainan and Yunnan ,for observing the reproductive ability of F1 hybrids.Ovarian nurse cell polytene chromosomes of F1 hybrid females were examined, to observe any synapsis in different zones of chromosomes.RESULTS: No embryo formation was found within the eggs produced by group Y♀×H♂, the hatching rate was zero. Low hatching rate was shown in other groups with (H♀×Y♂)F1, except for groups with (H×Y)F1×Y. Ovarian nurse cell polytene chromosomes from (H×Y)F1 hybrid females showed constant asynapsis at the 29th,36th and 37th zones in the chromosome 3R, and at the 4th and 6th zones in the chromosome X. CONCLUSION: Reproductive isolation did appear in An.minimus from Hainan and Yunnan.


STUDIES ON THE FEASIBILITY OF DETECTING CIRCULATING ANTIBODIES IN SALIVA OF SCHISTOSOMA JAPONICUM INFECTED RABBITS *

WANGZhaojun;LOUWenxian;ZHANGEnying;ZHANGXiangyan;XUEChunliang

1999, 17(4):  4-204. 

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　AIM: To investigate the feasibility of detecting anti- Schistosoma japonicum antibodies in saliva. METHODS: Saliva and serum samples of 5 infected, 7 reinfected and 8 treated rabbits were collected at different times periods. The CAb in saliva and serum was detected by using ELISA. RESULTS: The sensitivity of ELISA was 94.7% for saliva and 100% for serum. The specificity of ELISA was 100% for both saliva and serum. CONCLUSION: Saliva can be used to detect circulating antibodies for the diagnosis of schistosomiasis japonica.


EXPRESSION AND IDENTIFICATION OF RECOMBINANT 22.6 kDa FUSION PROTEIN OF SCHISTOSOMA JAPONICUM

SUChuan;MALei;WUHaiwei;SHENLei;CHENShuzhen;ZHANGZhaosong;WUGuanling

1999, 17(4):  5-208. 

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　AIM: To obtain a large amount of purified 22.6 kDa antigen of Schistosoma japonicum ( Sj 22.6) in large quantity.METHODS: The sequence of the gene fragment encoding Sj 22.6 was reformed by PCR and subcloned into plasmid vector pGEX-1λT that coded for the 26 kDa GST antigen of Schistosoma japonicum ( Sj 26 GST). The recombinant plasmid was transformed into E.coli TG 2 and then the positive recombinant clone was expressed by induction with IPTG. RESULTS: The recombinant Sj 22.6/ Sj 26 GST fusion protein was expressed in 5.1% of total bacterial protein and was easy to be purified with glutathione sepharose 4B. Moreover, the purified recombinant Sj 22.6 antigen could be cut off easily from the fusion protein with thrombin and had high immunogenicity. CONCLUSION: The purified recombinant Sj 22.6 protein and Sj 22.6/ Sj 26 GST fusion protein had the same immunological activity as the native Sj 22.6 kDa protein .


STUDIES ON THE CONTINUOUS CULTURE AND THE GROWTH OF ENTAMOEBA GINGIVALIS

CHENJinfu;LIUGuanyin;WENWangrong

1999, 17(4):  6-211. 

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　AIM: To establish a method for continuous culture of Entamoeba gingivalis(E.g. ).METHODS:The culture conditions of E.g. were compared by observing its size and survival time at room temperature. RESULTS: The growth of E.g. under different culture conditions including culture medium, temperature and pH were compared. The accompanying bacteria associate with E.g. FJ4 were isolated and identified. The average size of E.g. was 13.19 μm ～ 49.93 μm×9.88 μm ～ 30.74 μm. The optimal culture conditions of E.g. were:modified LES or YES medium, pH 6.4 ～ 6.7, nutritional liquids such as Lockes solution or yolk liquid with 20% bovine serum, penicillin, streptomycin and rice flour at 35 ℃.Reproduction of E.g. peaked at the fourth day of incubation,and the survival time of E.g. was 120 h～168 h. CONCLUSION: E.g. could be continuously cultured in modified LES or YES medium by inoculating once every four days.


STUDIES ON THE RELATIONSHIP BETWEEN THE INFECTION OF INTERMEDIATE HOSTS OF PARAGONIMUS AND ECOLOGICAL ENVIRONMENT

CHENGYouzhu

1999, 17(4):  7-214. 

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　AIM: To explore the relationship between the infection of the intermediate hosts of Paragonimus and their ecological environment.METHODS:Three different villages in Fujian Province were chosen to conduct the study.The infection of snails and crabs were surveyed.RESULTS:① In Ningyang Village, Wuyi mountain(stream type),the infection rate of the Potamon crabs (723.9/each) in section Ⅰ(rapid stream) was 5 times higher than that in section Ⅱ(rapid stream) 142.6/each. ② In Xikou Village, Mingching county( ditch type): The infection rate of the snails (Semisulcospira libertina ) to cercaria and the Potamon crabs were 4.2% and 1 738.3/each, respectively in 1996, being 28.9 and 8.5 times higher than those ( 0.2% and 204.7/each) in 1982.③ In Jishang Village, Jianou City,the infection rate of the snails ( Tricula ) and the Potamon crabs were 4.3% and 355.4/each respectively in ditch Ⅲ, 4.3 and 8.3 times higher than those (1% and 42.9%/each) in ditch Ⅱ.CONCLUSION: P.westermani distributed along the stream system, usually concentrating in the small, slow-running water bodies of the stream. P.skrjabini distributed in still smaller water bodies.The environment suitable for the breedings of the snails and crabs are also the suitable places for Paragonimus parasites to complete their life cycles.


FUSION EXPRESSION OF PORCINE IFN-γ AND CYSTICERCUS CELLULOSAE ANTIGEN cC1 IN ESCHERICHIA COLI CELLS

GUOYingjun;WUDan;SUNShuhan

1999, 17(4):  8-217. 

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　AIM: To construct an expression vector,including a chimeric cDNA of porcine IFN-γ and Cysticercus cellulosae antigen cC1. METHODS: DNA fragments of porcine IFN-γ and cC1 including linkers were generated by polymerase chain reaction(PCR).The recombinant vector pJLA-PRcC1 was constructed by inserting a chimeric cDNA of porcine IFN-γ and Cysticercus cellulosae antigen cC1,and transformed to E.coli XL-Blue. RESULTS: The recombinant vector was identified by restriction analysis.An inserted fragment about 1.5 kb could be found.After induction,a 52 kDa new protein band appeared in SDS-PAGE. CONCLUSION: The fusion expression of porcine IFN-γ and cC1 antigen in Escherichia coli cells is successful.\;


EXPRESSION OF SCHISTOSOMA JAPONICUN FATTY ACID BINDING PROTEIN GENE IN SILKWORM CELLS AND LARVAE *

LIUJinming;CAIXuezhong;LINJiaojiao;YANGGuanzhen;SHENXiaochuanFUZhiqiang;SHIFuhui;SHENWei;LIMing;YUANChunxiu;LIHao;CAIYoumin;WUXiangfu

1999, 17(4):  9-221. 

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　AIM: To express the fatty acid binding protein ( Sj14FABP) gene of Schistosoma japonicun in the silkworm cells and larvae. METHODS: A 600 bp DNA fragment containing Sj14FABP gene was cloned into baculovirus transfer vector of pBacPAK His1 to construct recombinant transfer vector Sj14-pBac PAK His1 . Coinfection was accomplished with this vector and Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA in BmN cells. The recombinant virus of Bm-Sj14 was screened using dot-blotting. The BmN cells and silkworm larvae were infected with Bm-Sj14 to express Sj14FABF gene. Western blotting and ELISA were used to identify the antigenicity of the recombinant protein. RESULTS: Sj14FABP gene was successfully expressed in the BmN cells and silkworm larvae infected with Bm-Sj14. The product was a 18 kDa fusion protein. The yield in BmN cells was about 100 μg/1×10 6 cells and 33 μg/ml cell supernatant. In silkworm larvae, the product yield was 4 mg/ml haemolymph as well as 4.6 mg/g silkworm tissue. The recombinant protein could be recognized by Western blotting and ELISA using the sera from mice immunized with SWAP. CONCLUSION: Sj14FABP gene has been successfully expressed in BmNPV system and the product has high antigenicity.


IMMUNOCYTOCHEMICAL LOCALIZATION OF TWO FACILITATED GLUCOSE TRANSPORTERS OF SCHISTOSOMA MANSONI IN THE TEGUMENT OF SCHISTOSOMA JAPONICUM

JIANGJiawan;ZHONGCisheng;QILing;YUYongfu

1999, 17(4):  10-224. 

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　AIM: To observe the distribution of Schistosoma mansoni glucose transport proteins, SGTP1 and SGTP4, in the tegument of Schistosoma japonicum . METHODS: The rapidly frozen fixation technique and ultracryomicrotomy were adopted for preparing ultrathin cryosections of S.japonicum . Anti-SGTP1 and anti-SGTP4 antibodies were used to localize the corresponding antigens in the tegument of adult S.japonicum by immunocytochemical technique. RESULTS: SGTP1 was localized on the basal membrane of the tegument and its infoldings, SGTP4 was localized on the apical membrane of the tegument and its invaginations of S.japonicum . CONCLUSION: The same localization for SGTP 1 and SGTP4 in the tegument of S.japonicum and S. mansoni exhibited apparent homology between SGTPs of the two schistosomes.


ESTABLISHMENT OF IMMUNOGLOBULIN M(IgM)\|IMMUNOSORBENT AGGLUTINATION ASSAY (ISAGA) FOR DIAGNOSIS OF TOXOPLASMOSIS

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1999, 17(4):  11-227. 

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AIM∶To establish an immunosorbent agglutination assay (ISAGA)for detection of IgM antibodies against Toxoplasma gondii. METHODS∶In the ISAGA,wells of microtiter plates were coated with anti\|human IgM antibodies and sealed with 1% bovine serum albumin.After the test sera were added and incubated,the plates were washed, T.gondii tachyzoite antigen suspension was added, and incubated overnight at 37℃. The ISAGA results were evaluated by compari...

ISOLATION AND QUANTITATION OF CHLOROQUINE-BINDING PROTEINS IN PLASMODIUM BERGHEI *

WANGQinmei;WANGMingjie;CHANGHuiling;YANGBin

1999, 17(4):  12-230. 

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　AIM: To isolate and quantitate chloroquine (CQ)-binding proteins in both CQ-sensitive (CS) AND CQ-resistant (CR) Plasmodium berghei . METHODS: P.berghei infected mice were each given ig CQ 400 mg/kg, respectively, 3 h later, the proteins of the parasites collected were separated using Ultrogel R AcA 34 gel column , followed by CQ extraction , the CQ concentration was determined by HPLC (extra standard). RESULTS: There were 58 protein peaks in the CS, all of which were bound with CQ,fraction Nos. 64～86 (peak 9) being the predominant, accounting for 16.4% of the total amount of CQ-binding protein. In the CR there were 40 protein peaks, among which 32 were bound with CQ, the predominant fraction Nos.60～83 (peaks 6,7),had a higher CQ-binding ability than CS. CONCLUSION: The distribution and amount of each CQ-binding protein peak in both CS and CR strains of P.berghei were demonstrated, exhibiting differences in the ability of CQ-binding not only between CS and CR strains but also within the same strain.


简报

EFFECT OF ARTEMETHER ON NUCLEOSIDE UPTAKE AND NUCLEIC ACID CONTENT IN SCHISTOSOMA JAPONICUM *

ZHAIZili;MEIJingyan;YOUJiqing;YAOMinyi;XIAOShuhua

1999, 17(4):  13-234. 

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　AIM: To observe the effect of artemether (Art) on nucleoside uptake and nucleic acid content in Schistosoma japonicum . METHODS: RNA and DNA contents of both male and female worms harbored in mice treated intragastrically (ig) with Art 300 mg/kg for 24 h or 48 h were determined, respectively. After in vivo drug treatment , the schistosomes recovered were in vitro maintained in drug-free medium containing [ 3H]adenosine, [5- 3H] uridine or [methyl- 3H]thymidine at a final concentration of 37 MBq/L or 74 MBq/L for 2 h or 4 h, the tritiated nucleoside uptake and incorporation into nucleic acid of schistosomes were measured. RESULTS: The RNA and DNA contents of female worms recovered from the host 48 h after dosing were markedly decreased by 51.6% and 23.5%, respectively,while the RNA content of male worms showed 42.4% reduction. When the above-mentioned schistosomes were in vitro exposed to the tritiated nucleoside for 2 h or 4 h, apparent decrease in tritiated nucleoside uptake with reduction rates of 35.2%～50.1% was seen in female worms. The incorporation of [methyl- 3H]thymidine into the female worm DNA 2 h after incubation was reduced by 71.4% while the incorporation of [ 3H]adenosine into the female worm RNA and DNA 4 h after incubation was reduced by 65.2% and 50.0%,respectively. CONCLUSION: Art exhibited an apparent effect on the nucleic acid metabolism in schistosomes, especially in female worms.


论著

EVALUATION OF IMMUNOCHROMATOGRAPHIC TEST IN THE DIAGNOSIS OF PLASMODIUM FALCIPARUM AND PLASMODIUM VIVAX

ZHENGXiang;TANGLinhua;XUYongxiang;MENGFeng;ZHUWeidong;GUZhengcheng;QIANHuilin

1999, 17(4):  14-236. 

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　AIM: To evaluate the effectiveness of immunochromatographic test(ICT) in detecting Plasmodium falciparum and Plasmodium vivax in malaria endemic areas. METHODS: ICT was used to detect P.falciparum and P.vivax among patients with fever in the outpatient clinics by comparason with thick blood smear method.RESULTS: The sensitivity of ICT to detect P.falciparum and P.vivax was 96.7% and 90.4%,respectively. The specificity of ICT was 98.6%,and the coincidence rate was 94.7%. There is no cross reaction between P.falciparum and P.vivax . CONCLUSION: ICT could detect P.falciparum and P.vivax simutaneously,being more rapid and simple than blood smear method.


实验研究

PROTECTIVE EFFECTS OF LEISHMANIAL ANTIGENS AGAINST LEISHMANIA INFANTUM INFECTION IN LAGURUS LAGURUS *

CHAIJunjie;KwangPooCHANG;ZUOXinping;YanLeiHouYanyanZhangSongRuziguliJIANGWeiZhangLanying

1999, 17(4):  15-240. 

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　AIM: : To determine the protective effect of leishmanial surface antigens against experimental visceral leishmaniasis in Lagurus lagurus caused by Leishmania infantum .METHODS: Recombinant surface protein (rGP63) and lipophosphoglycan (LPG) of Leishmania were used with Corinebacterium parum vaccine as ajuvant to immunize Lagurus lagurus against a challenge with virulent strain of Leishmania infantum . The efficacy of immunoprotection was observed.RESULTS: When challenged with up to 2×10 7 promastigotes, the number of LD on the liver printing sections in the rGP63+LPG+CP immunised animals was significantly decreased ,the parasite reduction rate being 89.79%. LPG+CP gave a parasite reduction rate of 60.6% and rGP63/β-galactosidase fusion protein +CP showed a parasite reduction rate of 42.45%.Purified rGP63 showed no protection. Immunization with rGP63+LPG+CP followed by challenge inifection with 1×10 6,5×10 6 and 1×10 7 promastigotes also showed significantly reduced infection rates.CONCLUSION: A combination of rGP63+LPG+CP antigens could provide significant immunoprotection against L.infantum challenge in L.lagurus .


防治经验

FIELD APPLICATION OF ORAL ARTESUNATE FOR PREVENTING SCHISTOSOMA JAPONICUM INFECTION *

XUMingsheng;ZHANGShiqing;LISiwen;WANGTianping;CHENJiran;OUNeng;FANGGuoren;WANGQizhi;LIJuntian;ZHANGXinsheng

1999, 17(4):  16-243. 

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　AIM: To assess the preventive effect of oral artesunate against S.japonicum infection. METHODS:Residents in two pilots in the schistosomiasis endemic regions, 562 cases in Yanghe pilot,Wangjiang County and 218 cases in Shashan pilot,Guichi City,Anhui Province, were selected for this study. The residents were divided into two groups.Group Ⅰ received artesunate 6 mg/kg once every 2 weeks for 4 times 2 wk after contacting with infested water from July to September in 1997.Group Ⅱ received the same dosage of placebo at the corresponding times.Four weeks after the last administration,stool examination using hatching method and Katos method was conducted to evaluate the effect.RESULTS: In Yanghe pilot,2 cases were hatching positive in the artesunate-treated group with an infection rate of 0.7%(2/273), while 11 cases were stool positive in placebo group with an infection rate of 3.8%( 11/289)and a mean EPG of 26.40±1.49.In Shashan pilot,all cases in artesunate-treated group were stool negative, while 7 cases were stool positive in the placebo group with an infection rate of 6.3%(7/111),and a mean EPG of 14.23±2.14.The protection rate of artesunate was 80.9% and 100%,respectively,in the two pilots. CONCLUSION:Artesunate can protect the residents from S.japonicum infection effectively.