Table of Content

30 December 2000, Volume 18 Issue 6

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论著

SEQUENCE ANALYSIS OF SSU rDNA VARIABLE REGIONS OF LEISHMANIAISOLATES FROM HILLY FOCI AND PLAIN FOCI OF CHINA

BULing-yi;HUXiao-su;JINGBao-qian;YITao-lin

2000, 18(6):  1-324. 

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　Objective] To analyze the sequence difference of the SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China. [Methods] Specific SSU rDNA fragments from nuclear DNA of five Leishmania species and isolates were amplified by PCR. The amplified DNA fragments were cloned into pGEM R-T Easy vector. The specific fragments were sequenced by the automated DNA sequencer. [Results] Sequence analysis showed that the amplified DNA fragments of five Leishmania species and isolates were all 392 bp in length, point mutations were located in the two unique sequence (UQ-Ⅰ and UQ-Ⅱ); L.d.SC10 and L.d.GS7 had two same point mutations in UQ-Ⅱ, only L.d.GS7 had one in UQ-Ⅰ; no insertion/deletion. [Conclusion] Sequence difference of the SSU rDNA variable region existed between Leishmania isolates from hilly foci and plain foci; The sequences of the SSU rDNA variable regions of L.d. SD2 isolate and L.infantum were identical.


ON MOLECULAR IDENTIFICATION AND TAXONOMIC STATUS OF ANOPHELES LESTERI AND ANOPHELES ANTHROPOPHAGUS IN CHINA (DIPTERA:CULICIDAE)

MAYa-jun;QUFeng-yi;CAOYu-cun;YANGBao-jin

2000, 18(6):  2-328. 

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　Objective] To clarify the taxonomic status of Anopheles lesteri and An.anthropophagus in China. [Methods] Using molecular identification (PCR assay and rDNA-ITS2 sequencing) to examine the field anopheline mosquito specimens from Liaoning and Shandong. According to the ITS2 sequences, molecular phylogenetic tree was made. [Results] According to the molecular identification, An.lesteri and An.anthropophagus were distributed both in Liaoning Province and Shandong Province. The length and GC content of rDNA-ITS2 sequence were 451 bp, 46 2% in An.lesteri (n=6), and 448 bp, 46 0% in An.anthropophagus (n=10), respectively. The ITS2 sequences from presentation sites were same in An.lesteri, while the intraspecies difference in An.anthropophagus was 0 88%. The specific difference between An.lesteri and An.anthropophagus was 25 7%. By analyzing molecular phylogenetic tree, the relationship between An.lesteri and An.sinensis, An.anthropophagus and An.liangshanensis was found to be closer. [Conclusion] According to the molecular identification, it was defined that An.lesteri and An.anthropophagus were sympatric independent species in China.


IMMUNE RESPONSE IN MICE INDUCED BY C TERMINAL ENCODING GENE OF PLASMODIUM FALCIPARUM HISTIDINE RICH PROTEIN 2

MIAOJun;LIXun;XUECai-fang;LIUZhong-xiang;WANGXian-feng;ZHENRong-fen

2000, 18(6):  3-332. 

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　Objective] To explore the humoral and cellular immune responses in mice to eukaryotic expression recombinant plasmid encoding histidine rich protein 2 (HRP-Ⅱ) of Plasmodium falciparum. [Methods] The start and stop codes were introduced into HRP-Ⅱ gene fragment, the reading frame and the position of start and stop codes in HRP-Ⅱ were identified by sequencing. HRP-Ⅱ fragment containing the start and stop codes was cloned into pcDNA3 1(-) to form pcDNA3 1(-)/HRP-Ⅱ. The BALB/c mice were immunized i.m. with the plasmids for 3 times in 3 weeks intervals. Two weeks after the last immunization, the sera and splenocytes were collected to investigate anti-HRP-Ⅱ antibodies by ELISA and the splenocytes proliferation response to HRP-Ⅱ. [Results] Sequence data show that the reading frame and the position of start and stop codes are correct. Restriction enzyme digestion indicated that the HRP-Ⅱ gene fragment containing start and stop codes was successfully cloned into pcDNA3 1(-). Mice raised significant anti-HRP-Ⅱ antibodies after pcDNA3 1(-)/HRP-Ⅱ immunization, and the splenocytes proliferated prominently when stimulated with HRP-Ⅱ protein. [Conclusion] Eukaryotic expression recombinant plasmid \{encoding\} HRP-Ⅱ gene can induce significantly humoral and cellular immune response in mice. HRP-Ⅱ gene may be a good candidate for P.falciparum blood-stage multiple DNA vaccine.


PREPARATION AND CHARACTERIZATION OF McAbs AGAINST LACTATE DEHYDROGENASE OF PLASMODIUM FALCIPARUM~+

WUYing-song;DONGWen-qi;LIMing;GAOYang;BIHui-xiang;LIYing-jie

2000, 18(6):  4-335. 

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　Objective] To prepare and characterize the monoclonal antibodies(McAbs) against lactate dehydrogenase of the Plasmodium falciparum(LDHp). [Methods] BALB/c mice were immunized with purified recombinant LDHp and McAbs against LDHp were prepared according to the protocol of hybridoma technique. The McAbs were characterized by ELISA and Western blot analysis. [Results] Two McAbs against LDHp antigen were obtained. Both McAbs were IgG 2b . The titer of two McAbs(2A5,1H10) in the ascites was \{1∶25 600\} and \{1∶12 800\}, and in supernatant were \{1∶512\},\{1∶256\} respectively. The result of ELISA indicated that two McAbs reacted only with P.falciparum, and did not react with normal human red blood cells, P.vivax, Toxoplasma gondii, Schistosoma japonicum. It is recognized 33 kDa protein which was defined as LDHp by Western blot analysis. [Conclusion] Two hybridoma cell lines secreting high titer of McAbs against LDHp with high specificity were established.


EFFECT OF ARTEMETHER ON PHOSPHOGLUCOMUTASE, ALDOLASE, PHOSPHOGLYCERATE MUTASE AND ENOLASE OF SCHISTOSOMA JAPONICUM HARBORED IN MICE

ZHAIZili;YOUJi-qing;GUOHui-fang;JIAOPei-ying;MEIJing-yan;XIAOShu-hua

2000, 18(6):  5-338. 

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　Objective] To study the effect of artemether (Art) on phosphoglucomutase(GPM), aldolase(ALD), phosphoglycerate mutase(PGM) and enolase(ENO) of Schistosoma japonicum harbored in mice. [Methods] Mice infected with S.japonicum cercariae for 4～5 wk were treated ig with Art 100 mg/kg or 300 mg/kg and killed 24 h or 48 h after treatment for collection of worms. The activities of GPM, ALD, PGM and ENO in female and male worms were measured by the formation of NADPH or consumption of NADH. [Results] After the worms were exposed in vivo to Art 100 mg/kg for 24 h, the GPM, ALD, PGM and ENO activities in female worms were significantly decreased by 15%, 19%, 50% and 46%, respectively, while in male worms only the PGM and ENO activities were markedly decreased by 22% and 32%, repectively. Following exposure of the worms to Art 100 mg/kg for 48 h, the GPM and ALD activities in male worms were also significantly reduced by 21% and 18%, respectively, while the activities of GPM, ALD, PGM and ENO in female worms and those of PGM and ENO in male worms declined progressively with time. After the worms were exposed in vivo to Art 300 mg/kg for 24～48 h, all the activities of the above-mentioned enzymes in female and male worms declined significantly in a time-related pattern. [Conclusion] Art showed an apparently inhibitory effect on GPM, ALD, PGM and ENO in female schistosomes.


CLONING AND SEQUENCE ANALYSIS OF RESA GENE FRAGMENT OF PLASMODIUM FALCIPARUM ISOLATE FCC1/HN

LIXue-rong;YUXin-bing;SHANZhi-xin;MAChang-ling

2000, 18(6):  6-342. 

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　Objective] To determine the nucleotide sequence of the 3′-termal of the RESA gene Plasmodium falciparum isolate FCC1/HN, and find out the differences of the sequences of RESA gene among isolate FCC1/HN,FC27,NF7 and Palo Alto. [Methods] 3′-terminal fragment of RESA gene of P.falciparum isolate FCC1/HN was amplified by PCR method, then was cloned into pMD18-T vector. The recombinant was screened and identified by BamHI+XhoI and PCR technique. The nucleotide sequence of the 3′-terminal of the RESA gene was determined by the dideoxy chain termination method. DNASTAR and BLAST software were used to compare and analyze the RESA gene sequences among the different isolates. [Results] The 3′-termal fragment of the RESA gene with about 846 bp was specifically amplified by PCR, the recombinant pMD18-T-RESA was successfully constructed. Different degrees of diversity of the RESA gene sequences were found among P.falciparum isolates FCC1/HN、FC27,NF7 and Palo Alto. [Conclusion] There were differences in the sequences of RESA gene among the P.falciparum isolate FCC1/HN and three other isolates (FC27,NF7 and Palto alto).


AMPLIFICATION, CLONING AND EXPRESSION OF A GENE ENCODING HEXOSE TRANSPORTER OF PLASMODIUM FALCIPARUM

WEIQuan-de;YUXin-bing;YELing;XUJin

2000, 18(6):  7-346. 

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　Objective] To amplify,clone and express of a gene encoding hexose transporter of Plasmodium falcipuram(PfHT1) from Southern China isolate FCC1/HN for studing the immune of recombinant which protective from malaria parasite infection. [Methods] Cultivation of P.falciparum isolate FCC1/HN in vitro; extraction of genomic DNA from FCC1/HN using the alkali specific cleavage method; PCR amplification of PfHT1 and cloning into eukaryotic expression vector, pEGFPN3. The recombinant was introduced into mammalian cells, HEPG2 by using liposome-mediated transfection. [Results] The gene encoding PfHT1 was specifically amplified from the genomic DNA of P.falciparum isolate FCC1/HN. The size of amplified fragment was 1 516 base pair. The eukaryotic expression recombinant, pN3-HT1 , was constructed and expressed steadily in the hepatocarcinoma cell lines, HEPG2. [Conclusion] The gene encoding PfHT1 was successfully amplified and cloned. The pN3-HT1/HEPG2 cell line was built for expressing fusion protein of GFP-HT1.


实验报道

ANALYSIS OF TRACE ELEMENTS IN LIVER, SPLEEN AND BRAIN OF RATS INFECTED WITH TOXOPLASMA GONDII

GENGZhi-hui;SHIYi;FANGYan-qiu;LIShu-hong;LIULi

2000, 18(6):  8-349. 

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　Objective] To study the level of five trace elements(Fe 2+ 、Cu 2+ 、Zn 2+ 、Ca 2+ 、Mg 2+ )in the liver, spleen and brain of the rats infected with Toxoplasma gondii. [Methods] 20 rats were randomly and equally divided into two groups: normal group and experimental group. On the 64th day after injection of 1.5×106T.gondii/2 ml, all of the rats were killed and the atomic absorption method were used. [Results] Compared with the normal group, the level of Cu 2+ in liver in experimental group was significantly reduced(P<0 01), and the amount of Fe 2+ of sick rats were higher than that of the normal(P<0 01). No differences in the content of Zn 2+ 、Ca 2+ 、Mg 2+ between the experimental and control group were determined; the level of Fe 2+ 、Cu 2+ in spleen in experimental group was significantly altered(P<0 01). The amount of Mg 2+ of sick rats were higher than that of the normal (P<0 01), and no difference on the content of Zn 2+ 、Ca 2+ between the experimental and control group were estimated;the level of Fe 2+ 、Cu 2+ 、Mg 2+ in brain in experimental group was significantly reduced(P<0 01).The amount of Ca 2+ of sick rats were higher than that of the normal(P<0 02).No difference on the content of Zn 2+ between the experimental and control group were estimated. [Conclusion] T.gondii infection might cause changes in trace elements in the liver,spleen and brain of rats.


防治经验

LONGITUDINAL OBSERVATION ON THE CONTROL OF INTESTINAL HELMINTHIASIS

WENLi-yong;XIAZhao-hua;YAOShang-ying;YANGJi-shun;CHENGGuo-qiang;SUYing-long;SONGChang-cun

2000, 18(6):  9-353. 

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　Objective] To search suitable measure for rapid control intestinal helminthiasis and long-term strengthen efficacy. [Methods] The treatment was taken in egg-positive population of intestinal helminthiasis in 1986～1988. The treatment was carried out only in the selected population in 1989～1992. No measure was taken in 1993～2000. [Results] (1) The prevalence rate of hookworm, Ascaris and Trichuris decreased to 3 2%,37 3% and 3 5% respectively after administration of albendazole twice a year for 3 years. (2) The prevalence rate of hookworm continued to decrease to 0 5% after treatment on selected population. (3) The prevalence rate and the intensity of hookworm has been less than 1% and 10/LPG for 8 years. No hookworm larvae had been isolated from the soil.[Conclusion] The hookworm transmission was effectively controlled in the study site.


SURVEILLANCE OF FILARIASIS IN SOME VULNERABLE AREAS IN GUANGXI AFTER FILARIASIS ELIMINATION

XIEZu-ying;PANShi-xian;LUXian-gang;MAIFu-zhen;LIAONing;HEWei-tao;YANGXiao-chun;YANGYi-chao

2000, 18(6):  10-355. 

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　Objective] To explore the measures for continuing surveillance of filariasis. [Methods] Selecting some vulnerable spots for focal surveillance, double-slide biood sampling for microfilaria examination, dissecting vectors for detecting the mosquitoes infected with filarial larva, using IFAT for detecting antifilarial antibody. [Results] 27 938 persons were examined for microfilaria and 4 454 mosquitoes were dissected for filaria larva, all were negative. 3 606 serum samples were examined for antifilarial antibody average positive rate was 1 35%(0 39%～4 97%). [Conclusion] The results of surveillance showed that the achievement of filariasis control in Guangxi after filariasis elimination is consolidated.


临床研究

CLINICAL ANALYSIS OF 36 CASES WITH AMEBIC LIVER ABSCESS

QINShu-lin;WANGAi-xia;SHENGRui-yuan;LIUZheng-yin

2000, 18(6):  11-358. 

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　Objective] To investigate the clinical features of amebic liver abscess, the causes of misdignosis and the effect of medical and surgical therapy on patient′s prognosis. [Methods] The clinical features of 36 patients with amebic liver abscess admitted from 1982 to 1997 in our hospital were retrospectively reviewed. [Results] The major clinical manifestations were: abdominal pain (86 1%), fever (86 1%),hepatomegaly with tenderness (83 3%) and right intercostal tenderness(58 3%). Leukocytosis was observed in 61 1%, and increased of ESR in 88 5% (23/28). Serologies against Entamoeba histolytica were noted in 92 6%. Ultrasonography showed single lesions in 75% and right-lobe involvement in 75%. All patients were treated with metronidazole and 27 patients received treatment with needle aspiration or draining at the same time. After treatment, 10 patients were cured, 25 patients were improved significantly and effective rate was 97 2%. One patient died of hepatic failure. [Conclusion] Medical therapy alone was excellent for small abscesses, while percutaneous needle aspiration or draining was a successful approach in patients with large abscesses.