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Table of Content

    30 June 2010, Volume 28 Issue 3
    论著
    Effect of Schistosoma japonicum Mr 22 600 Particulated-antigen onDendritic Cells and CD4+CD25+ Regulatory T Cells
    SHILei;ZHOUYing;WANGYong*;LIANGYue-jin;ZHANGZhao-song
    2010, 28(3):  1-165. 
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    【Abstract】 Objective To study the role of Schistosoma japonicum Mr 22 600 particulated-antigen on dendritic cells (DCs) and CD4+CD25+ regulatory T cells. Methods In in vitro experiments, DCs were pulsed with Sepharose 4B coupling rSj22.6/26GST and soluble rSj22.6/26GST, respectively. The surface molecules of DCs were detected by flow cytometry and the function of DCs was detected by mixed lymphocyte reaction. For the reduction of CD4+CD25+ T cells, DCs pulsed with Sepharose 4B coupling rSj22.6/26GST and soluble rSj22.6/26GST, respectively, were cocultured with CD4+ T cells isolated from the spleen cells. The percentage of CD4+CD25+ Foxp3+ T cells in CD4+ T cells was detected by flow cytometry. In in vivo experiments, BALB/c mice were immunized with Sepharose 4B coupling rSj22.6/26GST, Freund′s adjuvant emulsified rSj22.6/26GST, rSj22.6/26GST, Sepharose 4B, Freund′s adjuvant and PBS, respectively. The percentage of DCs in draining lymph nodes and the percentage of CD4+CD25+Foxp3+ T cells in spleen cells were detected by flow cytometry. To analyze the inhibitory roles of CD4+CD25+ T cells on CD4+CD25- T cells, CD4+CD25+ T cells were separated from mice immunized with Sepharose 4B coupling rSj22.6/26GST and Freund′s adjuvant emulsified rSj22.6/26GST, respectively, and cocultured with CD4+CD25- T cells. The proliferation of cells was assessed by [3H] thymidine incorporation methodResults In vitro, the expression rate of the surface molecules of CD40, CD80 and CD86 on the soluble antigen pulsed DCs were (43.5±6.2)%, (37.7±0.1)%, and (71.4±1.4)%, respectively. But on the Sepharose 4B coupling antigen pulsed DCs, they were (31.2±5.4)%, (32.0±1.6)%, and (63.8±1.0)%, respectively, which suggested that the Sepharose 4B coupling rSj22.6/26GST had less stimulating roles on DCs maturation. Furthermore, addition of DCs pulsed with Sepharose 4B coupling rSj22.6/26GST caused the expanding of CD4+CD25+ T cells. In vivo, immunization of Sepharose 4B coupling rSj22.6/26GST increased the number of CD4+CD25+ T cells. CD4+CD25+ T cells separated from Sepharose 4B coupling rSj22.6/26GST immunized mice had stronger inhibitory ability (cpm 1 420±335), compared with that of mice immunized with soluble antigen (cpm 3 558±147). Conclusion In contrast to the Freund′s adjuvant emulsified antigen, immunization with Sepharose 4B coupling rSj22.6/26GST increases the number of CD4+CD25+ T cells, which showed stronger inhibition on the CD4+CD25- T cell proliferation, and the mechanism of which may be involved in DCs maturation.
    Treatment of Rats Infected with Clonorchis sinensis Using Clinical Administration Regimens of Tribendimidine, Praziquatnel and Artesunate
    XUEJian;XULi-li;QIANGHui-qin;ZHANGYong-nian;XIAOShu-hua*
    2010, 28(3):  2-171. 
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    【Abstract】 Objective To evaluate the efficacy in treatment of Clonorchis sinensis-infected rats using the adminis-tration regimens of tribendimidine, artesunate and praziquantel applied in clinical treatment of clonorchiasis. Methods The doses of tribendimidine, artesunate and praziquantel used in clinical treatment of clonorchiasis were converted to the doses used in rats by the method of equal effective dose conversion among different animals, while the administration regimens of the drugs were designed basing on the regimens used in clinical trials. Thus, the following dose schedules were set up, i.e., tribendimidine 16 or 32 mg/(kg·d)×1, 2 or 3 d (bid), 8 or 16 mg/(kg·d)×3 d; artesunate 12 mg/(kg·d)×3 d(tid) and 16 mg/(kg·d)×3 d(bid); praziquantel 143 mg/(kg·d)×2 or 3 d (tid), 143 mg/(kg·d)×2 or 3 d(bid), 47.7 or 71.5 mg/(kg·d)×3 d. 151 rats were divided into 2 batches and each rat was infected orally with 50 metacercariae of C. sinensis. In the first batch of test, 79 rats were divided into 13 groups of 5-6 rats 5 weeks post-infection. Among them 6 groups were treated orally only with tribendimidine, artesunate or praziquantel, while other 7 groups were treated with tribendimidine combined with artesuante or praziquantel, or praziquatel combined with artesunate. The remaining 8 untreated rats served as control. In the second batch of test, 72 rats were divided into 13 groups of 5 rats. Among them, 7 and 6 groups were treated with tribendimidine and praziquantel, respectively, 6 weeks post-infection. The remaining 8 untreated rats served as control. Rats were sacrificed 14 days post-treatment, worms were recovered from the bile duct and the liver tissue. The mean worm reduction rate was calculated and compared among the groups by nonparametric method (Mann-Whitney test). Results In the first batch of test, the mean worm burdens in rats infected with C. sinensis and treated orally with tribendimidine 16 or 32 mg/(kg·d)×3 d(bid), praziquantel 143 mg/(kg·d)×3 d (tid), or 143 mg/(kg·d)×3 d (bid) were significantly lower than that of the control (P﹤0.01) with mean worm burden reductions of 94.2%-96.0%. No efficacy was seen when infected rats were treated orally with artesunate 12 mg/(kg·d)×3 d(tid). But in those treated with artesunate 16 mg/(kg·d)×3 d(bid), the mean worm burden was significantly lower than that of the control (P﹤0.05) with a mean worm reduction of 57.2%. In combined treatment, the infected rats treated with tribendimidine 16 or 32 mg/(kg·d)×3 d(bid) in combination with praziquantel 143 mg/(kg·d)×3 d(bid) or artesunate 16 mg/(kg·d)×3 d(bid), the difference of mean worm burden between each combined treatment group and control group was statistically significant (P﹤0.01) with mean worm reductions of 94.2%-99.4% which revealed that the worm reduction rate in combined treatment group was similar to the corresponding group treated with tribendimidine or praziquantel alone, but significantly higher than that of the group treated with artesunate alone. In infected rats treated with praziquantel 143 mg/(kg·d)×3 d(tid) plus artesunate 12 mg/(kg·d)×3 d(tid) or praziquantel 143 mg/(kg·d)×3 d(bid) plus artesunate 16 mg/(kg·d)×3 d(bid), the mean worm burden reductions were 93.6%-100%. In the second batch of test, the efficacy of tribendimidine obtained from infected rats treated with the drug 16 or 32 mg/(kg·d)×2 d(bid) and 3 d(bid), the difference of mean worm burdens between them was not statistically significant with mean worm reductions of 86.5%-95.1%. When rats were treated with tribendimidine 32 mg/(kg·d)×1 d(bid), the mean worm reduction was 73.0%, while the dose of the drug was given to the rats at 8 or 16 mg/kg daily for 3 days the mean worm reduction rates were 88.3%-92.6%. Treatment of praziquatnel 143 mg/(kg·d)×3 d (tid) resulted in a worm reduction of 96.9%, if the treatment course reduced to 2 d, the rate was 63.2%. Similar results were obtained in rats treated with praziquantel 143 mg/(kg·d)×2 d(bid) and 3 d(bid). Finally, administration of praziquantel at a daily dose of 47.7 or 71.5 mg/kg for 3 d exhibited no effect against C. sinensis. Conclusion When the dose schedules of tribendimidine, artesunate and praziquantel used in humans are converted to the doses for use in rats, tribendimidine and praziquantel exhibit satisfactory effect against C. sinensis, but artesunate shows no or less effect; the treatment course of tribendimidine can be reduced from 3 d to 2 d. Since tribendimidine and praziquantel used alone have endorsed high efficacy against C. sinensis in rats, combinations among the 3 drugs do not show better effect.
    Screening and Characterization of Peptides Specifically Binding to the Schistosomulum Tegument of Schistosoma japonicum
    WEIQi;LIUYan;ZENGQing-ren;CHENYu-xiao*;ZHOUJun;LIUBi-yuan;ZOUYan;YANGSheng-hui;CAILi-ting;LILi-xin;LANLing-mei
    2010, 28(3):  3-175. 
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    【Abstract】 Objective To screen and analyze the peptides in 12 phage-display peptide library specifically binding to the schistosomulum tegument of Schistosoma japonicum. Methods A 12 phage-display peptide library was screened with the S. japonicum schistosomula as the target cells for biopanning by degrees, positive clones picked randomly were deduced by DNA sequencing. According the sequencing result, immunohistochemical staining was performed to determine the specificity of the phages to the tegument. To test their targeting efficacy, the interested phage clones were infused back to the mice infected with S. japonicum, mice were sacrificed 2.5 hours later, and the phage distribution in the liver and the tegument of schistosomula was appraised, respectively. Results After 3 rounds of biopanning, the phage recovery rate increased from 0.77×10-8 to 0.75×10-5, indicating that the phage library was successfully enriched in the tegument of schistosomula. Seventy-five percent (15/20) of the analyzed sequences were identical with a sequence of QHPRIRKOOOOO. The immunohistochemical stainings showed this sequence specifically binding to the tegument. In vivo titering displayed that this sequence selectively targeted the tegument. Conclusion The peptide of QHPRIRKOOOOO specifically binds to the schistosomulum tegument.

    Protective Immunity of Cs-Rho GTPase RecombinantProtein Against Clonorchis sinensis Infection
    XIEHong-yan;HUXu-chu;XUJin;YUXin-bing*
    2010, 28(3):  4-179. 
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    【Abstract】 Objective To study the protective immunity induced by recombinant vaccination of Cs-Rho GTPase of Clonorchis sinensis(Cs). Methods 20 SD-rats(8 weeks) were divided into two groups: A(recombinant protein experiment group) and B (PBS control group). Rats in group A were immunized with l ml protein of Cs-Rho GTPase (90 μg/ml) and 1 ml Freund′s complete adjuvant through back and vola. 2 week later, the rats were given 1 ml protein of Cs-Rho GTPase (90 μg/ml) and 1 ml Freund′s incomplete adjuvant, followed by 1 ml protein of Cs-Rho GTPase (90 μg/ml) through intraperitoneal injection at 4, 7, 11 week after the first immunization. Rats in group B were given PBS in the same way as group A. All rats were challenged each with 50 Clonorchis sinensis metacercariae after the last immunization. 21 d later, fecal samples were collected from all rats for examining eggs (number of eggs per gram feces, EPG) in every 3-5 d. When eggs were found, the rats were sacrificed and worms were collected. IgG、 IgG1 and IgG2a in sera were detected by ELISA before every immunization. Mean number of worms and eggs, and antibody level in the experiment group were calculated and statistically compared with the controls. Results The mean number of worms and EPG were (9.2±9.9) and (956.8±1 062.5) respectively in group A, which were significantly lower than those of group B [(23.25±15.75) and (3 062.5±2 501.8) respectively] (P<0.05). The absorbency values of serum IgG (0.1, 0.45, 0.65, 0.6, 0.65), IgG1 (0.1, 0.45, 1.1, 1.0, 1.1), and IgG2a (0.1, 0.7, 1.1, 1.1, 1.1) before every immunization in group A were significantly higher than those of group B(almost always 0.1)(P<0.05). Conclusion Recombinant vaccination of Cs-Rho GTPase induces partial protective immunity against Clonorchis sinensis infection in rats.
    Cloning, Expression and Identification of EndothelialDifferentiation-Related Factor-1 Gene of Schistosoma japonicum
    DENGWang-ping;FENGZheng;XUBin;KONGJuan;HUWei*
    2010, 28(3):  5-184. 
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    【Abstract】 Objective To clone and express endothelial differentiation-elated factor(SjEDF)- gene of Schistosoma japonicum, analyze its immunogenicity and the stage-pecific expression at different developmental stages of S. japonicum. Methods Total RNA were extracted from eggs, cercariae, schistosomula and adult worms. The housekeeping gene SjActin was selected as the internal reference. According to the open reading frame for SjEDF- gene (GenBank accession number: AY336498), a pair of primers were designed to amplify the SjEDF- gene which was subc-oned into pET-8a vector. The recombinant plasmid SjEDF-/pET-8a was transformed into E. coli BL21 and induced with IPTG for expression. The recombinant protein was purified with Ni-TA resin. The immune rabbit sera was prepared by immunizing New Zealand white rabbits with purified recombinant SjEDF- protein. Western blotting was used to analyze the immunogenicity and the expression level of SjEDF- at the different developmental stages. Results The SjEDF- gene was detected with a band of 405 bp in eggs, schistosomula, female and male worms. The recombinant protein (rSjEDF-) was expressed as inclusion bodies (Mr 20 000). Western blotting analysis showed that the purified rSjEDF- protein was recognized by pooled sera of infected rabbits. The target protein was detected only in schistosomulum and adult worms. Conclusion The recombinant protein (rSjEDF-) shows certain immunogenicity, and is detected only in schistosomula and adult worms.
    Protective Efficacy Induced by Dendritic Cells Pulsed with GST in Combination with CpG Oligodeoxynucleotide against chistosoma japonicum Infection
    LIXiao-hong;CAOJian-ping;TANGLin-hua*;XUYu-xin;LIUShu-xian;WANGSheng-jun;CHENGJing
    2010, 28(3):  6-189. 
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    【Abstract】 Objective To study the protective effects on the infection of Schistosoma japonicum in C57BL/6 mice induced by dendritic cells DCs pulsed with GST in combination with CpG oligodeoxynucleotide. Methods GST was purified and used to stimulate DC2.4 cell line. Antigen loading was analyzed by immunofluorescence method. Thirty-five C57BL/6 mice were divided into seven groups(5 mice per group). Mice in groups A, B, C, D and E were immunized subcutaneously with DCs, DCs treated with PSA, DCs pulsed with GST, DCs stimulated with GST+CpG ODN, DCs stimulated with CpG ODN, respectively. For the above five groups, each mouse received 100 μl cell suspension at the density of 107/ml subcutaneously for three times at 2-week intervals. Each mouse of group F was immunized subcutaneously with 50 μg GST formulated in complete Freund′s adjuvant first, and 50 μg, 10 μg GST respectively in incomplete Freund′s adjuvant for the last two doses. Group G received PBS and served as control. Serum samples were collected 10 days after the final immunization, and were analyzed for specific antibodies by ELISA. At two weeks after the final immunization, each mouse were challenged by 30±1 cercariae of S. japonicum. Six weeks after infection the mice were sacrificed, and number of worms was counted. Results Light green fluorescence was observed in dendritic cells under the fluoroscope after pulsing with GST which indicated the protein loaded dendritic cells. The IgG level in groups C, D and F was 0.555 2±0.078 9, 0.715 0±0.052 3, and 2.127 0±0.411 5, respectively, all higher than that of group G(P<0.05). The worm reduction rate of group D was 53.3%, followed by group F (24.0%) and group C (21.3%). There was no significantly difference in the worm reduction rate between group D and groups F and C(P>0.05). Conclusion Dendritic cells pulsed with GST in combination with CpG oligodeoxynucleotide induce significant immunoprotection against the infection of Schistosoma japonicum.
    Expression of the Genes of Adenosine Deaminase, C-lectin andSerpin in the Salivary Gland of Aedes albopictus
    WUJia-hong;CHENGJin-zhi;CHENLu;SUNYu
    2010, 28(3):  7-193. 
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    【Abstract】 Objective To express the genes of adenosine deaminase(ADA), C-lectin and serpin (serine protease inhibitor) in the salivary gland of Aedes albopictus. Methods Total RNA was extracted respectively from salivary glands of unfed (group SG) and engorged adult female Ae. albopictus mosquitoes (group BSG), female carcasses without head and salivary gland (group C), and male bodies without heads but with salivary glands (group M). After the primers for the genes of ADA, C-lectin and serpin were designed respectively according to the reported Ae. albopictus gene sequences in GenBank, real-time fluorescent quantitative RT-PCR was performed to detect expression level of these genes in different tissues of Ae. albopictus using β-actin as internal reference. Results The mRNA expression level of ADA gene in the salivary glands from unfed adult female mosquitoes (group SG) was 545 and 123 times higher than those of female carcasses without head and salivary gland (group C) and male bodies without heads but with salivary glands (group M) (P<0.01). In group SG, C-lectin was 3 929 and 4 973 times higher than that in group C and M (P<0.01). High level of mRNA coding for serpin was detected in group SG, being 1 911 and 2 978 times higher than that in group C and M (P<0.01). There was no significant difference in ADA, C-lectin and serpin mRNA levels between unfed and engorged salivary glands (P>0.05). Conclusion ADA gene can be expressed in various mosquito tissues, but higher in salivary glands. The genes of C-lectin and serpin have been highly expressed specifically in salivary gland of female mosquito.
    Detection of Anisakid Nematodes by an SYBR Green Ⅰ Real-time PCR
    ZHANGShao-lei;WANGHe-lan;LIUJiang;NIFang;XUShi-san;LUODa-min*
    2010, 28(3):  8-199. 
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    【Abstract】 Objective To establish an SYBR Green Ⅰ real-time quantitative PCR method for the detection of anisakid nematodes with zoonotic potential from Taiwan Strait. Methods Anisakid larvae of six species (Anisakis sim-plex, A. physeteris, Raphidascaris trichiuri, Contracaecum aduncum, C. muraenesoxi, and Contracaecum sp., a predominant species in fishes in the strait) were obtained from the guts of marine fishes and identified chiefly based on the morphol-ogical features. The ITS-2 rDNA sequences from the larvae were amplified by PCR using universal primers, then cloned and bidirectionally sequenced. According to these sequences, six specific forward primers were designed and synthesized. Specificity was determined by a series of conventional PCR respectively, the ITS-2 sequences amplified above were cloned into T vector which was subsequently transformed into E. coli DH5α. Following extraction and identification, the positive recombinant plasmid was used as quantitative template to generate standard curve and melt curve. Sensitivity and reprod-ucibility were determined. Results All the 6 standard curves established by the recombinant plasmids showed adequate linear relationship between threshold cycle(Ct) and template concentration. Melt curves were specific and all the 6 corr-elation coefficients were above 0.998. In the reproducibility test, the coefficients of variation (cv) of Ct values for detection of the 6 nematodes ranged between 0.18% and 2.80%, and the cv of the inter-assay ranged between 0.55% and 1.94%. The sensitivity of the real-time PCR was 1×102 copies/μl, about 100 times higher than the conventional PCR assays. The real-time quantitative PCR detection needed only 3.5 hours from the sample treatment to result report. Conclusion An SYBR Green Ⅰ fluorescent quantitative PCR has been developed for detecting anisakid nematodes with adequate sensitivity and specificity.
    An Indirect ELISA for the Detection of Babesia caballiin Equine Animals
    GONGZhen-li;LIUGuang-yuan*;XIEJun-ren;CHAIHui-ping;ZHANGLi-yan;LIZhi-xin;TIANZhan-chen;WANGLu;LIUJian-gang
    2010, 28(3):  9-204. 
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    【Abstract】 Objective To clone and express BC48 gene of Babesia caballi, and to establish an indirect ELISA for the diagnosis of B. caballi in equine animals. Method The genomic DNA of B. caballi was extracted from the infected donkey blood. BC48 gene was amplified by PCR. The PCR product was cloned into expression plasmid pET-28a, and expressed in E. coli BL21 with IPTG induction. The recombinant protein was purified by Ni-NTA affinity chro-matography and was used as a diagnostic antigen to establish an indirect ELISA. The reaction conditions of the indirect ELISA were optimized. Specificity and sensitivity of this method were evaluated. Result BC48 gene of B. caballi was 1 272 bp. The recombinant protein was expressed in E. coli BL21 as a soluble protein with a molecular weight of about Mr 46 000 under induction of IPTG. The concentration of purified protein was 12.98 mg/ml. The best conditions were obtained for the ELISA when the antigen concentration was 65 μg/ml with the serum dilution of 1 ∶ 80. The protein specifically reacted with serum from donkey infected by B. caballi, but did not react with serum from donkey infected by Theileria equi (B. equi). Both ELISA and microscopy were applied to examine 17 donkeys in the field, 3 were positive by ELISA and 2 were found parasite-positive, respectively. Conclusion The indirect ELISA method may be used to detect B. caballi infection in equine animals.
    Sequence Analysis of Theileria annulata Surface Protein in Chinese Isolates
    DANGZhi-sheng;LUOJian-xun;LIUAi-hong;BAIQi;GUANGui-uan;LIUZhi-jie;MAMi-ling;LIYou-quan;GAOJin-liang;SUGIMOTOChihiro;YINHong*
    2010, 28(3):  10-209. 
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    【Abstract】 Objective To study the TaSP polymorphism in three Chinese isolates of Theileria annulata. Methods The isolates from Inner Mongolia Autonomous Region, Ningxia Hui Autonomous Region and Xinjiang Uygur Autonomous Region were cultured in RPMI 1640 medium. TaSP gene was amplified from genomic DNA extracted from schizonts using polymerase chain reaction (PCR) and sequenced. Its amino acid sequence comparison was carried out with Clustal W2 multiple sequence alignment program. Molecular component and motif prediction were performed with online servers. Results The comparison of TaSP amino acid sequences of the three isolates showed that the central region (aa position 38-161) predicted to be the highly immunogenetic domain was polymorphic both in size and amino acid sequence, while the N-terminal (first 37 aa) and C-terminal (last 154 aa) parts were strongly conserved. Phylogenetic analysis and percentage identity revealed that the Chinese isolates were closely related to the isolates from Turkey, but quite different from those of India, Morocco and Tunisia. More importantly, variability was noticed among Chinese isolates, which caused both the location and number′s differences of motif (casein kinase II phosphorylation sites) among three TaSP sequences. Conclusion TaSP polymorphism exists in the Chinese isolates of T. annulata.
    实验研究
    Molluscicidal Effect of Endophyte LL3026 from Buddleia lindleyanaagainst Oncomelania hupensis
    HANBang-xing;CHENJun*;HAOLei;ZHOUXiao-kun;HANFang-an;LILong-gen
    2010, 28(3):  11-213. 
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    【Abstract】 Objective To research the molluscicidal effect, active components, thermal stability and light stab-ility of endophyte LL3026 (Colletotrichum sp.) from Buddleia lindleyana. Methods The molluscicidal effect of LL3026 against Oncomelania hupensis was determined as referring to the WHO guidelines for laboratory molluscicidal test, and the control experiments were performed with 1 mg/L niclosamide or dechlorinated tap water. The active components from LL3026 were extracted by different polar solvents. The thermal stability and light stability of its extracellular moiety was examined at different temperature (30-100 ℃), different time (30-150 min) and different illumination time (1-9 d). Results Immersion test showed that the LC50 values for the LL3026 broth were 50.11, 3.43, and 1.55 mg/L for 24, 48, and 72 h, respectively. The ether extract of LL3026 broth showed the best molluscicidal activity compared with other fractions. Treated with 25 mg/L ether extract for 24, 48, and 72 h, the mortality of O. hupensis was 100%. The molluscicidal activity of LL3026 broth had no change at 80 ℃ for 120 min, and the snail mortality was 100%. A 48 h exposure to LL3026 broth which placed in an artificial climate box with 3 600 lx illumination for 9 d resulted in 86.7% snail mortality. Conclusion The fractions extracted from endophyte LL3026 isolated from B. lindleyana shows molluscicidal effect to O. hupensis.
    Recent Investigation and Treatment of 1 060 AdvancedSchistosomiasis Cases in Zhejiang Province
    ZHANGJian-feng;WENLi-yong*;ZHUMing-dong;YANXiao-lan;CHENWen;LILi;LINLi-jun;YULi-ling
    2010, 28(3):  12-217. 
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    【Abstract】 Objective To understand the current status of advanced schistosomiasis patients, deliver medical treatment, and improve the case management in Zhejiang Province. Methods The cases previously diagnosed or suspected as advanced schistosomiasis in the province were the subjects of the investigation. Questionnairing (demographical information, disease history, etc.), clinical examination (ascites syndrome, abdominal palpation), laboratory examination (blood biochemistry, anti-Schistosoma japonicum antibody, eggs), and ultrasonography of the abdomen were used confirming the diagnosis. Treatment was given to the patients. Results There were 1 060 advanced schistosomiasis patients in 32 counties of 7 prefectures in the province. Majority of them distributed in water network regions, and lived with poor economic conditions. The average age of the patients was (66.3±9.3), with 89.3% ranged from 50 to 80 years old. Clinically 71.3% of the cases were with splenomegaly, 27.6% with ascites, 0.9% cases of multiple granuloma in the colon and 0.2% cases with dwarfism. 1 023 patients (96.5%) had received medical treatments. 69.4% of the cases had serious complications with advanced schistosomiasis and 52.5% had concurrently disorders in other systems. 71.1% of the patients had subjective symptoms and 65.2% had hepatosplenomegaly with hepatic fibrosis and dysfunction. The serum positive rate of antiS. japonicum antibody was 15.7%. Stool hatching test and microscopy revealed no eggs in fecal samples, but 24 out of 38 cases were found metamorphic eggs by rectal biopsy. Three years′ medical treatment improved the clinical conditions in 74.3% of the cases. Conclusion Situation of the advanced schistosomiasis patients is quite critical. Treatment and care are urgently needed especially for those aged 50 to 80 years old.

    综述
    Progress in Development of New AntischistosomalDrugs in Recent Years
    XIAOShu-hua
    2010, 28(3):  13-225. 
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    【Abstract】 It is estimated that 200 million people have suffered the infection with schistosomes in the world. The fact that treatment of schistosomiasis only relies on a single drug praziquantel does not adapt to the demand. Although praziquantel exhibits excellent therapeutic efficacy, it shows no preventive action to the infection. Therefore, development of new anti-schistsomal drugs has been received serious attention. In this paper the recent development of new anti-schistosomal drugs such as oxadiazole-2-oxides, mefloquine, etc. has been reviewed, and the process of their development and anti-schistosomal properties recounted.
    Polysaccharide and Molecular Pathogenesis of Plasmodium falciparum
    ZHANGYan;YINJi-gang*;CHENQi-jun
    2010, 28(3):  14-230. 
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    【Abstract】 In the interaction of Plasmodium falciparum with human cells, sporozoite adheres to the receptor of the liver endothelial cell, then invades to liver. Merozoite binds to the surface of red blood cells, and invades to erythrocyte. The adhesion of membrane protein of the infected erythrocytes to the surface molecules of vascular endothelial cell in the vital organs leads to the obstruction of blood circulation eventually. The adhesion is mediated by interaction between parasite derived ligands and the negative charged polysaccharides on the surface of host cells. This review is to discuss the molecular mechanisms in the host-parasite interactions.
    研究简报
    Protein Analysis of Trichomonas vaginalis Treated by Pulsatilla chinensis In vitro
    YANYan;GAOXing-zheng
    2010, 28(3):  15-233. 
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    【Abstract】 Water extract of Pulsatilla chinensis (PWE) (0.625 mg/ml) was added to the medium for the culture of Trichomonas vaginalis. After 2 h and 4 h treatment, the content and component of soluble proteins in the parasites were analyzed by SDS-PAGE. A total of 28 protein bands was demonstrated. After treated with PWE for 2 h, the content of 5 protein bands increased and that of 4 bands decreased significantly. Four hours later,a new protein band appeared in the electrophoretogram, and the content of 5 bands increased and that of other 5 bands decreased.
    The Accessibility for Anthelmintic Administration by DifferentSupply Patterns in Mass Deworming
    ZHANGQian;LIHua-zhong*
    2010, 28(3):  16-236. 
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    【Abstract】 Questionnairing was performed in selected fields. Data were collected including the number of people who were getting/taking medicines, reasons for not getting/taking medicines, and a comparison was made on the rates of getting/taking drugs among three patterns of medicine supply. In the mode of drug-receiving at public expense, the rates of getting drugs and taking drugs were 98.4% and 94.1% respectively in Hainan province, 93.2% and 90.9% in Guizhou province. In the mode of drug-delivery at public expense, the rates of getting drugs and taking drugs were 97.5% and 96.4% in Hainan province, 98.8% and 96.5% in Hunan province. In the mode of drug-purchase at people′s own expense, the rates of getting drugs and taking drugs were 96.8% and 95.6% in Guizhou province, 94.4% and 91.5% in Sichuan province. The reasons for not taking drugs included forgetting, drug-losing or unwell condition. Three patterns of drug supply can get satisfactory results through careful arrangement. The government can choose an appropriate pattern suited to local conditions.
    Clinical Analysis of 16 AIDS Patients Complicated withToxoplasmic Encephalitis
    ZHAOJian-rong;ZHANGSha;DUJian-ping;WANGShu;WANGDa-yong
    2010, 28(3):  17-238. 
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    【Abstract】 Sixteen AIDS patients complicated with toxoplasmic encephalitis (TE) were retrospectively analyzed between August 2008 to August 2009 with a mean age of (37.0±11.6) years. The most common clinical symptoms were headache (68.8%, 11/16) and fever (62.5%, 10/16), and 6 with Babinski sign (37.5%). 81.3%(13/16)were with CD4+ cells<200/mm3. Both sera and CSF showed 62.5% (10/16) TOXO-IgG positive by ELISA. CT and MRI scan demonstrated bilateral and multiple lesions with marked peripheral edema effect, and an enhanced scanning showed small finger ring as the major feature. 15 patients got improved by either oral sulphadiazine tablets or sulphadiazine tablets plus clindamycin capsule, 10 cases received combined HAART treatment, and 1 case died with septic shock.
    Construction of Full-length cDNA Library ofErythrocytic Stage Plasmodium vivax
    FANGQiang;XIAHui;YUANYuan-ying;CAOJun;WANGXue-mei;QIWen-juan;GAOQi*
    2010, 28(3):  18-封三. 
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    【Abstract】 Blood samples were collected from vivax malaria patients without antimalarial drug therapy. After filtration through Plasmodipur filter to remove white blood cells, Plasmodium vivax-infected RBCs were enriched by Percoll. Total RNA of P. vivax in red blood cells was isolated. A full-length cDNA library of erythrocytic stage P. vivax was constructed by the SMART cDNA library construction kit. The volume and recombinant rate of the library were evaluated. The inserted fragments were identified by PCR amplification. The titer of cDNA library was 1.14×106. The length of inserted fragment ranged from 900 to 2 500 bp, and the recombination efficiency accounted for 97.2%.
    病例报告
    Two cases of suspected cystic echinococcosis in spermatic cord
    LVSheng-fang;MAOXue-ying;KongHai-chao
    2010, 28(3):  20-171. 
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    A case of amebic cerebral abscess postmortem diagnosed
    WANGWei-wei;LIFei-hong;ZHAOYing-hai;DAIShi-zhong
    2010, 28(3):  20-189. 
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    Liposomal amphotericin B cured two cases of visceral leishmaniasis tolerant to sodium antimony gluconate
    YUANQiao-rong
    2010, 28(3):  21-213. 
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    Cerebral infection of sparganosis:a case report
    HUANGChun-chen;LILin-xin;DONGQiang;CAOWen-jie*
    2010, 28(3):  22-233. 
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    Schistosome eggs in cystic lymph node
    XIANGJin-jian;TIANFu*;LIMing-zhong;JIANGXue-feng;DENGShi-hong;HUXiao-miao;ZHANGBo-tao;LUOFei
    2010, 28(3):  23-236. 
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