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Table of Content

    30 April 2002, Volume 20 Issue 2
    论著
    Functional Characterization of the 5′ Proximal Flanking Fragment of Plasmodium falciparum GBP130 Gene
    WANGXian-feng*;MIAOJun;LIUZhong-xiang;LIXun;ZHENRong-fen;XUECai-fang
    2002, 20(2):  1-71. 
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     Objective To identify the putative regulation elements with strength- and stage- specificity in 5′ proximal flanking sequence of P.falciparum GBP130 gene. Methods Plasmids containing different deletions of upstream of the GBP 130 promoter were constructed. For strength-specific analysis, pGBPCATΔ2, pGBPΔ2/400 and pGBPΔ2/800 were electroporated into ring-stage P.f. respectively, and the expression level of CAT in each plasmid was detected by liquid scintillation counts(LSC). For stage-specific analysis, transfectants with pGBPΔ2/400 and pGBPΔ2/800 were harvested at 5 hours post-transfection(h), 15 h and 46 h respectively, and the CAT expression levels were detected. Results In strength-specific analysis, the expression level of CAT in pGBPΔ2/800 and pGBPCATΔ2 was similar, down-regulated significantly in pGBPΔ2/400. The CAT level showed significant difference between pGBPΔ2/400 and the control. In stage-specific analysis, the CAT level of pGBPΔ2/400 was higher than that of pGBPΔ2/800 at the time point of 5 h, and lower at 15 h and 46 h. Conclusion This strength-specific promoter activity was due to the difference of 5′UTR length:the longer the 5′UTR the higher the promoter strength, and two poly (dA∶dT) tracts in the proximal sequence could enhance the promoter activity. The length of 5′UTR regulated the promoter activity in a stage-specific manner. The shorter 5′UTR was functional at ring stage, while the longer one prompted transcription at trophozoite and schizont stage. The functional role of poly (dA∶dT) tracts in stage-specific regulation of GBP130 remains unclear.
    Cloning of a Species-specific Gene Fragment from Cryptosporidium parvum and the Development of Diagnostic PCR Primers
    TIANZong-cheng;ZHANGXi-chen;LIJian-hua;YINJi-gang;YANGJu;HEHong-xuan
    2002, 20(2):  2-75. 
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     Objective To develop a pair of diagnostic PCR primers for Cryptosporidium parvum . Methods A species-specific gene fragment of C.parvum was obtained through RAPD analysis. After the fragment was isolated,purified,cloned and sequenced,a pair of primers FF was designed and synthesised based on the sequence. With the primers, the anticipated fragment in size of 603 bp was amplified by PCR from 2 American strains and 4 Chinese strains of C.parvum . The samples of 35 rabbits feces and 55 human feces were detected by PCR with primers FF and 021,the latter was a species-specific diagnostic primer reported by Morgan. Results All six strains amplified by the primers FF showed same detection rate with 021. Sensitivity test indicated that DNA of 1 oocyst per gram of feces could be detected by the PCR. Conclusion The primers FF showed high specificity and sensitivity, and can be used for diagnosing Cryptosporidium parvum infection.
    Construction and Expression of an Eukaryotic Recombinant Plasmid from a Hybrid Gene of Antigen for Plasmodium falciparum
    YAOLang*LIYing-JieLIMing
    2002, 20(2):  3-78. 
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     Objective To construct an eukaryotic recombinant plasmid with HGFSP, a hybrid gene encoding the antigen epitopes of MSA1, MSA2, RESA and CSP in different developing stages of Plasmodium falciparum(P.f. ). Methods HGFSP was sub-cloned into an eukaryotic expression plasmid pcDNA3 from a prokaryotic recombinant plasmid pSK-HGFSP to construct the eukaryotic recombinant expression plasmid pc-HGFSP. The identified recombinant was then transfected into HepG2 cells with liposome-mediated method. The G418-selected positive cell clones were tested to identify the immunogenicity of HGFSP-expressing antigen. Results It was evidenced that HGFSP was correctly inserted into pcDNA3 by restriction enzymes map analysis. HGFSP-expressing antigen-specific fluorescent response was observed in pc-HGFSP-transfected HepG2 cells. The results of SDS-PAGE and Western-blotting showed that there was a 23 kD protein band, which can be specifically recognized by anti-sera of HGFSP-expressing protein in pc-HGFSP-transfected HepG2 cell lysis. Conclusion Pc-HGFSP, an eukaryotic recombinant plasmid encoding hybrid antigen epitopes of P.f ., was constructed successfully and the antigenicity of pc-HGFSP-expressing protein was confirmed.
    Study on Genomic Diversity among Different Isolates of Schistosoma japonicum in China
    QIUChi-ping;ChrisSpolsky;XIAMing-yi;CHENQin;TONGXiao-mei;GONGXin;GeorgeDavis;FENGZheng
    2002, 20(2):  4-82. 
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     Objective To study genomic diversity among the isolates of Schistosoma japonicum in China. Methods Schistosome adults were collected from different endemic areas. Mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome C oxidase 1 (CO1) gene fragments of the worms were amplified by PCR, cloned into plasmid, and finally sequenced. Program MEGA was used for sequence analysis. Results The ND1 and CO1 genes amplified from different geographic strains were 476 bp and 1 033 bp, respectively. There are two distinct haplotypes,type Ⅰ and Ⅱ, for both ND1 and CO1 nucleotide sequences. The differences between the two types were 4.0% and 3.4%. They are considered to be two different genetypes by the phylogenetic analysis. In individual mitochondrial gene, type Ⅰ of ND1 was fixedly accompanied with type Ⅰ of CO1 gene, and type Ⅱ of ND1 with type Ⅱ of CO1. Conclusion There are two different genetypes of ND1 and CO1 genes of S.japonicum in China.
    In vitro Antimalarial Effect of Daphnetin Relating to Its Iron-chelating Activity
    MULing-yun;WANGQin-mei;NIYi-chang**
    2002, 20(2):  5-85. 
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     Objective To investigate the in vitro antimalarial effect of daphnetin relating to its iron-chelating activity. Methods Schizontocidal activity of daphnetin and desferrioxamine B was tested through an in vitro assay based on the routine in vitro cultivation of Plasmodium faciparum FCC1 strain. The iron-chelating ability of each was measured by the fluorescent probe calcein. Results Daphnetin exhibited a modest iron-chelating ability compared with the powerful iron-chelator desferrioxamine B. In vitro test at the range of 0-12 μmol/L daphnetin showed a dose-dependent schizontocidal activity which could be inhibited if mixed with Fe 2+ in a ratio of 2∶1. Conclusion The dose-dependent antimalarial activity of daphnetin is related to its iron-chelating activity.
    Sequence Analysis of rDNA-LSU Gene of Orientobilharzia turkestanicum from Mainland of China
    ZHANGGuang-jun;CHENQin;QIUChi-ping;XIAMing-yi**
    2002, 20(2):  6-89. 
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     Objective To classify the taxonomic status of O.cheni in relation to O.turkestanicum var. tuberculata from the mainland of China by comparing their nucleotide sequences of nuclear ribosomal partial large subunit gene (LSU). Methods The genomic DNA of adult worms were extracted by the GNT-K method. The target gene was amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid PCR-blunt (Invitrogen). Recombinant plasmids were amplified in E.coli , extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of O. turkestanicum was retrieved from GenBank and aligned with our data in BioEdit. Results The nucleotide sequences of LSU between O.turkestanicum var. tuberculata and O.cheni was 100% identical, and 99.99% identical between O.turkestanicum var. tuberculata and O.turkestanicum . Conclusion This study demonstrated high similarity in LSU nucleotide sequences, and the results do not support O.cheni as an independent species. O.cheni may be a synonym of O.turkestanicum var. tuberculata , and O.turkestanicum var. tuberculata is probably also a synonym of O.turkestanicum .
    Study on Low Density Lipoprotein Receptor of Adult Schistosoma japonicum
    WANGLiang*;LIZhuo-ya;WANGYi;HEAi;XIEMin-hui;ZHANXi-mei
    2002, 20(2):  7-93. 
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     Objective To provide a theoretical basis for the study of vaccines against Schistosoma japonicum , the receptor for human LDL in the tegument of adult Schistosoma japonicum was investigated. Methods Proteins existed in adult Schistosoma japonicum membrane were extracted by Triton X-100 and purified through reverse-phase high performance liquid chromatography (HPLC),and the main protein peaks were then collected separately. 125 I-LDL of human plasma as the ligand, through the radioautography and radioligand binding assay, the protein which can bind human serum 125 I-LDL specifically was identified. The molecular weight and IEF were detected by SDS-PAGE. Results According to the radioautography and radioligand binding assay, a protein with retention time of 10.5 min was proved to be able to bind human serum 125 I-LDL specifically. SDS-PAGE revealed that the molecular weight of the purified protein is 60-65 kDa, and its IEF is 6.7. Conclusion LDL binding protein may exist on the surface of both male and female adult Schistosoma japonicum with the function of obtaining cholesterol from host circulating system.
    Assessment of Therapeutic Efficacy of Chloroquine Against Falciparum Malaria in an Outbreak Area in Xishuangbanna,Yunnan
    ZHANGZai-xing;LIChong-zhen;HUANGGuo-zhen;YANGYa-mingZHOUSheng;SUNXiao-dong;LILi
    2002, 20(2):  8-97. 
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    Objective To investigate the failure of treatment with chloroquine in Yunnan in order to help formulate adequate antimalarial drug policy. Methods A World Health Organization 28-day in vivo test on therapeutic response for uncomplicated falciparum malaria in area with low or moderate transmission was adopted. Patients of age ≥ 6 months old were admitted without limitation in density of parasitaemia and body temperature. Clinical and parasitological observation was conducted for patients on day 0, 1, 2, 3, 4, 7, 14, 21, 28 and 35. Results Of 62 patients identified as malaria cases infected by Plasmodium falciparum only, Plasmodium vivax only or by both species, 52 cases infected by Plasmodium falciparum only were included in the study. The overall treatment failure rate was 40.7%, with early treatment failure (ETF) rate of 1.8% and late treatment failure rate (LTF) of 38.9%. Conclusion The treatment failure rate was much higher than the rate of 25% recommended by WHO. It is suggested that use of single chloroquine should be stopped in the treatment of falciparum malaria cases in such area. No relationship was found between the failure rate and the density of malaria parasites.
    Study on Number and Mature Rate of Eggs in Gravid Proglottids of Taenia solium
    MAYun-xiang;SUYun-pu;YANQiu-ye;HELi-jun;YANXu-xia;CHENJian-she;FENGShi-xian;LIUHui
    2002, 20(2):  9-100. 
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     Objective To investigate the number and mature rate of eggs in gravid proglottids of Taenia solium . Methods Ten worms of Taenia solium , expelled from patients, were detected. Eggs were collected from the last 10 gravid proglottids of each worm. Results & Conclusion The egg number in each mature proglottids varied from 3 900 to 126 520, and the mean number was 28 332. The mature rate of eggs was from 7.00% to 36.00% with an average of 29.12%, which was lower than that in proglottids naturally excreted with feces. With suitable temperature and humidity, the proglottids developed continually after excreted out of host body. Two to three days later, the mature rate of their eggs increased to 85%-90%.
    Sensitivity of Oncomelania Snail to Niclosamide in China
    DAIJian-rong;ZHOUXiao-nong;LIANGYou-sheng;ZHANGYan-ping;JIANGYu-ji;XIWei-ping;HUANGYi-xin;CHENChang;HUANGMing-xi;ZHUYin-chang
    2002, 20(2):  10-105. 
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     Objective To understand the variation in response of Oncomelania hupensis to niclosamide. Methods Snails were collected from 37 sampling areas distributed in 10 provinces (municipalities) using random environmental sampling methods in accordance with the different types and categories of snail habitats. In laboratory the snails were immersed in solutions of niclosamide for 24 and 48 hours at 25℃. Results 1.0 mg/L niclosamide showed 100% killing effect on snails in 24 hours. The LC 50 concentrations for snails immersed for 24 hours ranged from 0.0320 to 0.1689 mg/L with a mean value of 0.0920 mg/L. 0.5 mg/L niclosamide showed 100% killing effect on snails in 48 hours. The LC 50 values for snails immersed for 48 hours ranged between 0.0299 and 0.1114 mg/L with a mean of 0.0627 mg/L. There is a significant difference in snail sensitivity to niclosamide between sampling areas. Conclusion The sensitivity to niclosamide varied in snails from different sampling fields, but the chemical in a concentration of 1.0 mg/L showed 100% effect of killing snails, which is consistent to the manual of schistosomiasis control.

    Prevalence Survey on Cyclospora cayetanensis and Cryptosporidium ssp. in Diarrhea Cases in Yunnan Province
    ZHANGBing-xiang;YUHui;ZHANGLi-li;TAOHong;LIYan-zhong;LIYing;CAOZhi-kuan;BAIZhi-ming;HEYong-qing
    2002, 20(2):  11-108. 
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     Objective To investigate the prevalence and distribution characteristics of Cyclospora cayetanensis and Cryptosporidium ssp. infection in diarrhea cases of Yunnan Province. Methods To collect fresh faeces from diarrhea cases in 7 counties/cities, examine the specimens by direct smear with iodine staining and modified acid-fast staining. Results The infection rate of C. cayetanensis and Cryptosporidium ssp. was 3.97% and 5.29%, respectively. The infection rate of the two pathogenic coccidians was as high as 10.64% and 8.51% in preschool children. C. cayetanensis was found in 3 counties and Cryptosporidium in 6 counties. Conclusion Both C. cayetanensis and Cryptosporidium ssp. are prevalent in Yunnan Province with the latter distributed more widely, and the two pathogens are more prevalent in preschool children.
    临床研究
    Evaluation of Gd-DTPA enhanced MR in the Diagnosis of The Degeneration Stage of Cerebral Cysticercosis
    ZENGFei-yan;WANGChang-xin
    2002, 20(2):  12-111. 
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     Objective To detect the value of qualitative and orientating diagnosis in the degeneration stage of cerebral cysticercosis with Gd-DTPA enhanced MR. Methods Sixty-nine cases of cerebral cysticercosis were diagnosed by enhanced MR as degeneration stage, and confirmed by immunological examination and/or by surgery. MR plain scanning was conducted for the same cases. Results The plain scanning showed single or multiple lesions with long T1 and long T2 signals, and the enhanced scanning showed nodular or annular lesions. The diameter of the lesion after enhanced scan was not more than 22mm with an average value of 8.1mm. Some cases showed single lesion on plain scanning but showed multiple lesions after enhanced scan. Conclusion The enhanced MR shows more typical features of the degeneration stage cerebral cysticercosis. It can define the number, position and range of the lesions, and can improve the accuracy of differential diagnosis, and therefore be of importance in formulating treatment scheme and prognosis.