Table of Content

30 June 2003, Volume 21 Issue 3

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论著

Efficacy of Naphthoquine, Artemisinine and a Combination of the Two Drugs in the Treatment of Falciparum Malaria

WANGJing-yan;SHANCheng-qi;FUDa-dong;SUNZhi-wei;DINGDe-ben

2003, 21(3):  2-133. 

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　Objective To compare the efficacy and safety of naphthoquine, artemisinine and a combination of the two drugs in the treatment of faciparum malaria. Methods Of 230 patients, 100 patients were treated with combined regime (Co-NQ), 100 patients were treated with naphthoquine (NQ) and 30 patients were treated with artemisinine (QHS). All patients were hospitalized for 7 days and followed up for 28 days. Results The mean fever clearance time for Co-NQ, NQ, and QHS was (17.5 ±12.3)h, (32.7±17.7)h and (18.1±9.7)h respectively; the mean parasite clearance time was (30.0±8. 8)h,(45.5±10.0)h and (29.1±6.0)h respectively; and the 28 days cure rate was 97.0% ,100.0% and 66.7% respectively. Conclusion The Co-NQ possesses benefits of both naphthoquine and artemisinine, acting rapidly, with a short course of only one dose and a high cure rate. The regime is well tolerated by patients.


Dynamic Observation on Ultrasonographic Image During Chemotherapy of Hepatic Cystic Echinococcosis and a Proposal for Ultrasonic Classification

CHAIJun-jie;Monhebat;Yerjian-SULITAN;JINGQiang;CHANGQing;ZHANGHe-ling

2003, 21(3):  3-139. 

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　Objective To submit a proposal for the classification of ultrasonographic image in patients with hepatic cystic echinococcosis, on the basis of characteristic and distribution of ultrasonic image of hydatid cysts in cases and systematic observation on dynamic changes of sonographic image under treatment with emulsion albendazole and in relation to clinical efficacy. Methods The ultrasonic image of 645 cysts in 497 cases with liver cystic echinococcosis was classified. The distribution of different types of image in patients was analyzed. A comparative analysis of correspondence between the sonographic type and clinical efficacy was made in association with the rule of changes in ultrasonic features during chemotherapy. Results The ultrasonographic image of hydatid cysts was divided into six types. The distribution of ultrasonic types in patients reflected the process of natural evolution of cysts in human bodies. The dynamic change of ultrasonic image during treatment was identical with the natural evolution process of hydatid cysts. Conclusion The proposal for ultrasonographic classification submitted reflects a long evolutionary process from growth to death of hydatid cysts in human body. The dynamic change of hydatid cysts during chemotherapy indicated that the effect of drug has accelerated this process. Therefore this classification may be applied in diagnosis and to judge the chemotherapeutic efficacy of cystic echinococcosis.


Expression of Nitric Oxide Synthase NOS mRNA in the Gonad of Oncomelania hupensis Cultured in Different Temperature

YANGKun;ZHOUXiao-nong;YUChuan-xin;YINXu-ren;HONGQing-biao;SUNLe-pingYANGGuo-jing;ZHANGYan-ping;HUANGYi-xin

2003, 21(3):  4-143. 

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　Objective To study the expression of nitric oxide synthase (NOS)mRNA in the gonad of Oncamelania hu-pensis in different temperature. Methods The snails were cultured at temperature of 0℃ , 15 ℃and 25℃for 1 month. The total RNA of each group was extracted with the RNA extraction kit. A pair of degenerate primers was designed from conserved regions of mammalian NOSs, and the expression of NOS mRNA in the gonad was measured by means of reverse transcription polymerase chain reaction (RT-PCR). Results The target genes of NOS were detected in the gonad of snails. The level of expression of snail NOS mRNA in 25℃ group and 0℃group was significantly higher than that in the control group( P <0.01), there was no significant difference for the expression products between 15℃group and control group (P>0.05). Conclusion The designation of primers of the snails was validated. The impact of temperature on the expression of snail NOS mRNA was determined, which suggested that NO plays an important role in the physiological and pathological modulation.


Cloning and Identification of the Gene Fragments of Paragonimus westermani

LINGJia-jian;HOUMin;LIUJian-nan;ZHANGZi-hao;ZHANGYao-juan

2003, 21(3):  5-146. 

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　Objective To screen and identify the recombinants from the cDNA library of the adult Paragonimus west-ermani (PwA) for immunodiagnosis and immunoprophylaxis. Methods PwA cDNA library was screened with the PwA antigen immunized rabbit sera(IRS) pre-absorbed by the extract of E. coli XLl-Blue. The recombinants from positive clones were amplified by PCR, sequenced and cut off by KpnI/BarnHI and, then sub-cloned into pRESETB vector. The fusion protein was expressed,analysed by SDS-PAGE and identified by Western blotting with immune rabbit serum against worm antigen of Paragonimus westerrnani. Results The inserted cDNA fragment from the positive clone Pw-2 was about 800 bp, which contained an open reading frame(ORF) encoding Pw pre-procathepsin L belonging to cysteinase family. Expression product of Pw-2 was a fusion protein of 32 kDa, which can be recognized by immune rabbit serum against worm antigen of Paragonimus 晈esterrnani. Conclusion A recombinant plasmid Pw-2 encodes Pw pre-procathepsin L is constructed.


Studies on Fractionation and Protective Immunity of the Membrane Antigen from Hepato-portal Juveniles of Schistosoma japonicum

WANGMin;YIXin-yuan;ZENGXian-fang;ZHOUDong-ming;ZHANGShun-ke;ZHANGJie;YUANShi-shan

2003, 21(3):  6-149. 

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　Objective To explore the immunological characteristics of the membrane antigen from hepato-portal juveniles of Schistosoma japonicum and its protective immunity against S. japonicum (Sj) in mice. Methods Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and enzyme-linked immune electro-transfer blot(EITB) methods were used to recognize the membrane antigens from hepato-portal schistosomula (SjHmAg) by infected rabbit sera (IRS) and normal rabbit sera (MRS). Kunming mice were immunized subcutaneously three times(0,2,4weeks) with SjHmAg. Each mouse was challenged with 40±1 cercariae. Six weeks later the mice were killed, worms and liver eggs were counted. Results 7 major protein bands appeared on SDS-PAGE. IRS mainly reacted specifically with SjHmAg of 23, 33 and 63 kDa. Compared with the control groups, the reduction rate of worms and eggs per gram liver in the experimental group were 16.2% and 54.4 % , respectively. Conclusion Different protein components from SjHmAg are obtained using SDS-PAGE, and the antigen can induce a protective immunity against Sj in mice.


Expression of IL-2 and TNF-a in the Liver and the Effect of Injection of These Cytokines on Liver Fibrosis in Mice Infected with Schistosoma japonicum

ZENGLing-lan;LUODuan-de;LIShu-li;LIUWei;HEYong-wen

2003, 21(3):  7-153. 

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　Objective To detect the expression of IL-2 and TNF-α in the liver at different period postinfection of Schis-tosoma japonicum and their effect on liver fibrosis after supplementary injection of these cytokines. Methods Mice were infected with schistosome cercariae and divided into 3 groups. Two groups were injected (ip) every other day with IL-2 and TNF-α respectively for consecutive 4 wk. The third group and an uninfected group of normal mice were regarded as control. The ABC immunohistochemistry and pathologic image multimedia quantification system were applied to detect activity of IL-2 and TNF-α. Results The level of IL-2 and TNF-α in the liver in infected but untreated group slowly decreased (from 8, 11, 14 to 18 wk). The supplementary injection of the cytokines at 6 wk postinfection in the two groups increased the cytokines significantly, the level of IL-2 or TNF-αwas higher at 1 - 8 wk after the last injection than that of both infected and uninfected control groups (P<0.01). The granulomatous inflammation and fibrosis in the livers of the two groups were slighter than that of the control. Conclusion At the 6th wk postinfection with egg deposition, exogenous supplementation with TNF-αor IL-2 induces enhanced expression of the two kinds of cytokines, corresponding to a diminished degree of the liver granulomatous inflammation and fibrosis.


Cloning and Sequence Analysis of a Novel Stage-Specific cDNA from Adult Trichinella spiralis

FUBao-quan;WANGFeng;WUXiu-ping;NIUTing-xian;LUQiang;LIUMing-yuan;PascalBoireau

2003, 21(3):  8-156. 

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　Objective To clone a stage-specific novel cDNA from 5 day-old adult worm (ADS) of Trichinella spiralis. Methods The cDNA library of AD5 was screened by an AD5 stage-specific cDNA probe labeled with digoxigenin (DIG). The positive clones were sequenced and analysed. Results The positive clone contained a cDNA insert of 1 132 bp in length with a full length open reading frame (ORF) of 1 032 bp. The cDNA encoded a polypeptide of 343 amino acid residues(aa) with a molecular weight of 35.1 kDa and an isoelectric point (IP) of 4.8. InterProScan analysis showed that the 117 - 120 aa (SGYG) was a glycosaminoglycan attachment site, 27- 86 aa was nematode cuticle collagen N-terminal domain and 153-228 aa was collagen repeat (G-x-y) domain. Signal PV2.0 analysis indicated that the region of 1-43 aa was a singal peptide. Blastn homology analysis in Genbank revealed that the cDNA had no obvious homology to any other known gene sequence. Blastp analysis revealed high homology to cuticle collagen with identities more than 40 % . Conclusion A novel ADS stage-specific cDNA encoding a full length ORF was cloned and sequence analysis showed this gene encoded cuticle collagen of Trichinella spiralis.


Effect of Daphnetin on SOD Activity and DNA Synthesis of Plasmodium falciparum in vitro

MULing-yun;WANGQin-mei;NIYi-chang

2003, 21(3):  9-159. 

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　Objective To investigate the effect of daphnetin on superoxide dismutase (SOD) activity and DNA synthesis in P. falciparum in vitro. Methods The effect of daphnetin, daphnetin-Fe complex and desferrioxamine B on SOD activity of P. falciparum (P. f) FOCI in vitro was determined with a SOD test-kit. The level of DNA synthesis of P. f synchronized cultured in vitro at various developmental stages after treatment of daphnetin or desferrioxamine B was assayed by fluorescein Hoechest 33258. Results The total SOD activity decreased by 60% after daphnetin treatment while it only decreased by 22% if treated with desferrioxamine B. No effect on SOD activity of P. f treated with daphnetin-Fe complex was observed. The level of DNA synthesis of P. f trophozoites in synchronized in vitro culture was significantly lower than that of the control. Conclusion Daphnetin lowered SOD activity and decreased DNA synthesis of P. f in vitro.


cDNA Cloning and Sequence Analysis of Major Allergen of Dermatophagoides farinae Derf2 in South China

HAOMin-qi;XUJun;ZHONGNan-shan

2003, 21(3):  10-163. 

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　Objective To clone and analyze the cDNA of major allergen Der f 2 of Dermatophagoides farinae in south China. Methods cDNA of Der f 2 was cloned by RT-PCR, screened and their sequences were analyzed. Results cDNAof Der f 2 was cloned. The sequence of the cloned Der f 2 was different with that published (D10448) in GenBank, with 87 additional nucleotides inserted into the 62th nucleotide of the original one. According to the original ORF, the deduced amino acids that were located prior or after the inserted 29 amino acid sequence showed no changes. Conclusion The cDNA of Der f 2 was cloned from Dermatophagoides farinae and its sequence showed significant difference with that reported in the GenBank.


Application of Multivariate Regression in Analyzing Factors of Schistosomiasis Japonica Transmission in Poyang Lake

WUWei-ping;LINDan-dan;HUFei;GUANYa-yi;WangYan-An;ZHUHong-qing;CAOChun-li;CHENHong-gen

2003, 21(3):  11-166. 

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　Objective To analyze factors affecting transmission of schistosomiasis japonica in Poyang Lake. Methods Successive surveillance data for at least three years from 1992 to 1998 at the schistosomiasis endemic administrative villages surrounding Poyang Lake were collected, including the egg positive rate by stool examination, investment on treatment of patients and mollusciciding in snail habitat, risk areas, bovine infection rate. Data on rainfall and temperature were also obtained from the relevant agencies. Step-wised regression method was employed to analyze the data. Results The regression model established is statistically significant, R2 equals 0. 735, P < 0. 01. The accepted variables affecting the transmission of the disease were natural logarithm of human infection rate in last year, risk areas, infection rate of bovine, investment values of niclosamide per risk area and value of praziquantel administered per infection rate. Conclusion Chemotherapy and mollusciciding effectively reduced schistosomiasis transmission in Poyang lake region, while the infection rate, risk areas, bovine infection rate still drive the transmission.


Experimental study on Hybridoma Cells Agglutination Test for Diagnosis of Schistosomiasis Japonica in Mice

LIUWen-qi;LIYong-long;AndreasRuppel

2003, 21(3):  12-169. 

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　Objective To detect the infection of Schistosoma japonicum in mice with a novel test based on agglutination of hybridoma cells and to study the mechanism of the hybridoma cells agglutination. Methods The procedure was developed with a murine cell line H226 producing a monoclonal antibody specific to schistosome 31/32 kDa antigen and sera collected from mice infected with different numbers (10,30,50) of S. japonicum cercariae in different period. Immunofluorescent test was carried out with the hybridoma cells and schistosome-infected sera. Results The circulating antigen was detected by the test as early as 2 weeks after a heavy infection and all mice showed positive results in the test by 5 weeks after infection. The titers of antigen rose along with the lime post infection, and the tilers of sera from heavy infection were statistically higher than that from the mice receiving a lower number of cercariae. Specific yellowish green fluorescence appeared on the membrane of the hybridoma cells; no signal was detected inside. Conclusion Hybridoma cell agglutination test (HCAT) may become useful to diagnose schistosomiasis.


实验报道

Cloning and Sequence Analysis of Eg95 cDNA from Different Stages of Echinococcus granulosus in Xinjiang

LINRen-yong;DINGJian-bing;WENHao;ZHANGWen-bao;LIJun;LUXiao-mei

2003, 21(3):  13-172. 

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　Objective To study expression and sequence differences of Echinoaxcus granulosus 95(Eg95) antigen cD-NA from different stages of protoscolex, oncosphere and adult worm of E. granulosus from Xinjiang Uighur Aut. Reg. Methods In accordance with the sequence of Eg95 antigen cDNA, the primers of Eg95 were designed. Eg95 antigen cDNAs were amplified by PCR from protoscolex, oncosphere and adult worm cDNA libraries of E. granulosus, respectively and were cloned into pUCm-T plasmid, and sequenced. The sequences were analyzed by DNAman and GenBank/BLAST biosoftware. Results PCR results showed that Eg95 antigen cDNA was amplified from three stages of E. granulosus cDNA libraries. Sequencing analysis indicated that the Eg95 cDNA length was 402 bp, same as the reported data in GenBank. Conclusion The Eg95 antigen cDNA was expressed in the different life-cycle stages of E. granulosus in Xinjiang and there was no nucleic acid sequence difference of Eg95 antigen among the protoscolex, oncosphere and adult worm of E. granulosus.


Expression of Matrix Metalloproteinase 2 and 9 in Liver Tissue of Patients with Schistosomiasis Japonica

TAOJun;CAIWei-min;CHENJia-lin;ZHANGYan-ping;WENGHong-lei;LIURong-hua

2003, 21(3):  14-175. 

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　Objective To explore the change of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9 (MMP-9) expression in liver tissue of the patients with schistosomiasis japonica. Methods Liver biopsy materials were examined pathomorphologically in 26 patients with schistosomiasis in advanced stage and 5 cases without the disease as control. The expression of MMP-2 , MMP-9 and C-Ⅳwere studied by immunohistochemistry, and result was analyzed by picture quantitative analysis technique. Results Immnoreactive MMP-2 was mainly expressed in the plasma, the membrane of hepato-cytes and hepatic sinusoids, it was also found in myofibroblast cells and endothelial cells of blood vessel. Immnoreactive MMP-9 was observed in hepatic stellate cells, sinusoids and myofibroblast cells, sometimes it was also seen in the plasma of hepato-cytes and epithelial cells of bile duct. The expression of MMP-2, MMP-9 and C-IV was significandy stronger in patients with advanced schistosomiasis than that of the control(P<0.05). The expression of MMP-2 was relevant to the inflammation grade and stage of liver fibrosis(P<0.01). The expression of MMP-9 did not show significant change(P>0.05) and the expression of C-Ⅳ was consistent with that of MMP-2. Conclusion The study indicates that increased expression of MMP-2 participated in the initiation and development process of liver fibrosis, MMP-9 was related to the initiation of liver fibrosis and pathological change existed in hepatic sinusoids in advanced schistosomiasis japonica.


Preliminary Study on Primary Culture of Cells from Oncpmelania hupensis

PENG-Yan;JIANGMing-sen;ZHONGQin-ping;GUIJian-fang;DONGHui-fen

2003, 21(3):  15-178. 

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　[Abstract] Objective To study the primary culture of cells from Oncomelania hupensis. Methods After washed in the sterile solution with antibiotic, the Oncomelania hupensis snails and their eggs were dissected. The soft tissue, liver, mantle and the embryo were collected and torn up respectively. Above tissues except mantle were digested by a mixture containing equal volumes of 0.25% trypsin and 0.02% EDTA for several hours at 4 1C . The cells after digestion were inoculated. Meanwhile, the tissue of mantle were inoculated by moist system method. All cells were cultured separately in medium 1/2 RP-MI1640 containing 20% calf serum and antibiotics (penicillin G 100 lU/ml, streptomycin 100 fig/ml) at pH 7.2 - 7.4 and temperature of 27 t - 28 "C . Results Affter the embryonic tissue was digested by tryspin/EDTA mixture, lots of dissociated cells were obtained. Some of the cells began to adhere to the culture flask surface after cultured for 5 days. The adhering cells were flat and polygonal, about( 15-20 x 12-15)fun in diameter. But most of the cells were still suspending in the media. The suspending cells were round, usually about 8-12 fim in diameter with a few reaching 30 - 35 /um in diameter. These cells grew well and could be subcultured. Conclusion Embryonic cells from Oncomelania hupensis can be primarily cultured and subcul-tured.