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Table of Content

    30 December 2009, Volume 27 Issue 6
    述评
    Malaria Situation in the People’s Republic of China in 2008
    ZHOUShui-sen;WANGYi;FANGWen;TANGLin-hua
    2009, 27(6):  1-457. 
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    In total, 26 873 malaria cases and 108 594 suspected cases with 23 deaths were reported by the annual case reporting system in 858 counties of 22 Provinces/Municipalities/Autonomous Regions (P/M/A) in 2008, and the annual incidence was 0.21/10 000. Through the internet reporting system 26 358 malaria cases were reported from 994 counties of 30 P/M/A. The number of malaria cases and the rank of P/M/A were basically in concordance in the two systems.
    Among the 858 counties with reported malaria cases, 11 counties with an incidence more than 10/10 000 distributed in Yunnan (3 counties), Hainan (2), Anhui (4), Guizhou (1) and Tibet (1). There were 76 counties in which the malaria incidence was between 1/10 000 and 10/10 000.
    1 034 falciparum malaria cases accounted for 3.8% of the total cases, of which 75.4% (780) were imported cases from 207 counties/cities of 17 P/M/A. Indigenous falciparum malaria was found in 27 counties/cities of Yunnan and Hainan Provinces, of which 19 counties/cities were in Yunnan (5 less than that of 2007), and 8 counties/cities in Hainan (also 5 less than that of 2007).
    Focal outbreaks occurred in 93 villages of 10 counties in Yunnan, Anhui, Zhejiang and Hubei Provinces. 266 malaria cases resulted from the outbreaks, which accounted for 1.0% of the total reported cases.
    Although a considerable decrease in malaria incidence contributed to the implementation of the National Malaria Control Program and the Global Fund Programme, Yunnan and Hainan Provinces were still the relatively high transmission areas. Yunnan ranked No.2 in the country in terms of the case number, while Hainan ranked down to No.3 by malaria incidence in 2008. 5 871 malaria cases were reported from the two provinces in 2008, accounting for 21.9% of the total reported cases in the country. There were 4 027 cases with 6 deaths reported from Yunnan, the incidence was 1.16/10 000, a reduction of 52.5% than that in the last year. Among the reported cases, 678 were falciparum malaria with 67% imported cases. The number of reported cases in Hainan was 1 844, with an incidence of 2.18/10 000, a reduction of 46.2% than the last year.
    In central China, the re-emergence of malaria was considerably rolled back in 2008, but Anhui Province was still the No. 1 in the country either in the number of malaria cases or the incidence. 13 476 malaria cases were reported from Anhui in 2008, accounting for 50.1% of the total cases in the country, with an incidence of 2.42/10 000, decreased by 51.6% in comparison to that of 2007. The number of reported cases in Henan Province was 3 044, decreased by 25.8% in incidence. Hubei Province reported 1 088 malaria cases with an incidence of 0.18/10 000, decreased by 41.9%. 668 cases were reported from Jiangsu Province, decreased by 28.9% in comparison to that of 2007.
    Cases reported from other P/M/A occupied about 10.1% of the total. Over a hundred cases were reported from each of Guizhou, Zhejiang, Shandong, Guangdong, Sichuan, and Shanghai. Less than 100 cases were reported in each of Guangxi, Fujian, Chongqing, Hunan, Jiangxi, Liaoning, Shaanxi, Shanxi, Gansu, and Tibet in 2008.
    In summary, the re-emergence of malaria has been basically rolled back through several years′ efforts, but malaria has still been an important problem of public health in China, especially in the southern and central parts. Yunnan and Hainan still faced a critical situation of malaria endemics with the persistence of Plasmodium falciparum, especially imported malaria in the border areas. In the central parts of the country, particularly in Anhui, the malaria incidence was pressing with the highest incidence and majority of malaria cases in 2008. In addition, provinces not covered by the Global Fund Program, such as Guizhou, Zhejiang and Shandong, are confronting an increasing malaria burden, which may become a new challenge to the National Malaria Control Program.
    论著
    Prokaryotic Expression, Identification and Histolocalization of the Taenia asiatica Enolase Gene
    DUWu-ying;DAIJia-ling;HUANGJiang*;HUXu-chu;XUJin;YUXin-bing;LIAOXing-jiang;LANGShu-yuan
    2009, 27(6):  2-462. 
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    Objective To express enolase gene of Taenia asiatica, investigate the immunoreactivity of the recombinant TaENO pretein, and immuno-histolocalize the presence of the recombinant TaENO in adults of T. asiatica. Methods The gene encoding enolase of T. asiaticaTaENO) was cloned by high throughput sequencing from the cDNA library of adult T. asiatica. The coding region of TaENO was amplified by PCR, and cloned into a prokaryotic expression vector pET-30a(+). The recombinant plasmid was transformed into E. coli BL-21/DE3 and followed by expression of the protein induced by IPTG. The protein was purified by Ni-IDA affinity chromatography, and tested by SDS-PAGE. Its immunoreactivity was examined by Western blotting. The mice were immunized subcutaneously with purified TaENO formulated in Freund’s adjuvant. Serum samples were collected and analyzed for specific antibodies by ELISA. The localization of TaENO in adult worms was demonstrated by immunofluorescent technique. Results The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant TaENO was about Mr 47 000 with a concentration of 0.37 mg/ml. It was recognized by antisera of SD rats immunized with TaENO, sera of taeniasis patients and sera of infected swine. The immunofluorescence assay revealed that TaENO immune serum located in the tegument of T. asiatica adult. Conclusion The TaENO gene has been expressed with immunoreactivity, and the recombinant TaENO is immunolocalized in the tegument of T. asiatica adult.
    Inhibition of Plasmodium yoelii Circumsporozoite Protein on the Activation of Nuclear Transcription Factor in Hepatoma Cells Stimulated by TNF-α
    DINGYan;CHENJi-de;ZHOUTao-li;FUYong;PENGXiao-hong;XUWen-yue*
    2009, 27(6):  3-466. 
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    Objective To investigate on the effect of Plasmodium yoelii(BY265 strain)circumsporozoite protein (CSP)on the activation of nuclear transcription factor κB(NF-κB)in hepatoma cells(HepG2)stimulated by TNF-α. Methods Entire coding sequence of CSP was reverse transcribed and amplified by RT-PCR with sporozoite total RNA as template,then cloned into pFLAG-CMV8 for construction of the recombinant plasmid pFLAG-CMV8-CSP. Indirect immunofluorenscence staining with rabbit anti-csp was applied to verify the expression and distribution of FLAG-CSP fusion protein in HepG2. Three groups were established for the experiment: group A with HepG2 transfected by pFLAG-CMV8 as negative control, group B with HepG2 transfected by pFLAG-CMV8 and stimulated by 100 ng/ml TNF-α, and group C with HepG2 transfected by pFLAG-CMV8-CSP and stimulated by 100 ng/ml TNF-α. Dual-luciferase assay and EMSA were performed to detect the nuclear translocation and activation of NF-κB, to observe if pFLAG-CMV8-CSP suppressed the activation of NF-κB in HepG2 stimulated by TNF-α. Result The expression of pFLAG-CMV8-CSP was localized on cytoplasm of HepG2. The activity ratio of firefly luciferase to Renilla luciferase in group C(0.228±0.029) was significantly lower than both groups A(0.438±0.085)and B(0.571±0.030) (P<0.05). EMSA showed that the band in group C was significantly weaker than group B. Conclusion Plasmodium yoelii CSP localizes in the cytoplasm of HepG2 and inhibits the activation and nuclear translocation of NF-κB in HepG2 stimulated by TNF-α.
    Epidemiological Survey on Clonorchiasis sinensis in an Endemic Area of South Hunan Province
    DUANJi-hui;TANGXiao-yu;WANGQiao-zhi;TANGYang;ZHANGZong-si;LIZheng-xiang;LIUAi-hua;WUYan-jun;CHENWen-hua;HUANGQi-rong
    2009, 27(6):  4-471. 
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    Objective To make an epidemiological investigation on clonorchiasis sinensis and relevant factors in the south of Hunan Province. Methods One village from each of Lengshuitan District and Qiyang County was selected for the survey from November to December in 2006. Stool samples from villagers were collected and examined by modified Kato-Katz method. Questionnairing was performed for relevant knowledge and attitude among residents. The infection rate in animal reservoirs and intermediate hosts were detected. Results A total of 586 cases with Clonorchis sinensis infection were found from 777 people with a prevalence of 75.4%. The average egg density was 451 eggs per gram(EPG) feces. Light, moderate and heavy infections occupied 85.5%(501/586), 14.0%(82/586),and 0.5%(3/586)respectively. Prevalence in males and females was 76.9% (316/411) and 73.8% (270/366) respectively with no significant difference(χ2=1.013, P>0.05). Infections were found in all age groups, with the highest prevalence in the group of 70 to 79 years(85.7%, 30/35). By occupations, the prevalence was 82.5%(447/542)in peasants, 79.3%(42/53)in doctors, 73.7%(28/38)in teachers, and 73.5%(25/34)in local cadres. The infection rate was 17.4%(29/167) and 7.4%(2/27)in Parafossarulus striatulus and Alocinma longicornis, and 69.2%(9/13)and 5.3%(1/19) in Carassius auratus and Cyprinus carpios respectively. Adult worms were found in all 3 dogs dissected. Over 80% inhabitants did not know that this disease can be acquired by eating raw fish. 95.6%(153/160)of the farmers and 56.7%(349/616)of the students had a history of eating raw fish. The water was contaminated with C. sinensis eggs by using untreated feces as fertilizer for farming and by scrubbing pail latrines in the ponds. Conclusion The prevalence of clonorchiasis in human population is high in Lengshuitan District and Qiyang County of Hunan Province. Eating raw fish and using untreated feces as fertilizer are the most important epidemiological factors of the disease.
    Fleas on Small Mammals in the Surrounding Area of Erhai Lake
    DONGWen-ge;GUOXian-guo*;MENXing-yuan;GONGZheng-da;WUDian;ZHANGZheng-kun;ZHANGLi-yun
    2009, 27(6):  5-475. 
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    Objective To investigate the distribution pattern,species diversity and community structure of fleas on small mammals in the surrounding area of Erhai Lake, and the relationship between fleas and their hosts. Methods Different geographical areas surrounding the Erhai Lake in Yunnan were selected as investigated spots. Small mammals were captured with baited cages. The cage-traps were examined and re-baited each morning. All fleas on the hosts were collected and identified. The richness (S), evenness (J′), diversity index (H′), dominance index (C′), total ectoparasite infestation rate (Rpt), total ectoparasite infestation index (Ipt), and constituent ratio (Cr) were used to measure the community structure. Results Althgether, 3 303 small mammals and 3 243 fleas were collected. From the 21 species of small mammal hosts, 13 species of fleas were identified. In southern area of the Lake, the species richness (21 species of small mammals & 12 species of fleas) was highest among the three selected areas. Seventeen species of small mammals and 8 species of fleas were found in eastern area, and only 13 species of small mammals and 7 species of fleas found in the west. This implied the probable influences of ecological environments on the fleas and their corresponding hosts. The community structure of fleas on small mammals was complex. The species diversity, species composition, community structure and distribution pattern of fleas were simultaneously influenced by the hosts' body surface microenvironment and the macroenvironment (habitat). Conclusion The fleas are commonly distributed in small mammals in the areas and their communities are related to host species and the habitats.
    Immune Protection of Tegument Protein rSj29 against Schistosoma japonicum in Mice
    CHENHong;FUZhi-qiang;CHENLei;QIUChun-hui;FUGuang-wei;LIYe;SHAODong-hua;FENGXin-gang*;LINJiao-jiao
    2009, 27(6):  6-482. 
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    Objective To clone, express and characterize a tegument protein gene of Schistosoma japonicum (Sj29), and investigate the immune protection of the recombinant protein against S. japonicum in mice. Methods The gene coding for Sj29 protein was amplified by PCR, and the sequence was analyzed by bioinformatics tools. Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+). The recombinant plasmid was transformed into E. coli BL21(DE3)and induced the recombinant with IPTG. The recombinant protein(rSj29)was purified by His-binding-resin affinity chromatography and characterized by Western blotting. Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval)respectively with 100 μl recombinant rSj29(0.1 mg/ml), adjuvant or PBS. At the 15th day after the final inoculation, each mouse was challenged by 40±2 cercariae of S. japonicum. At the 53th day after infection, the mice were sacrificed to obtain the number of adult worms, number of eggs in liver and feces. Serum samples were collected at pre-immunization and certain time after immunization, and were analyzed for IgG by ELISA. The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique. mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR. Results A 576 bp Sj29 gene fragment was obtained. The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body. The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice. The number of adult worms(15.4±5.9), number of hepatic eggs(40 143.3±2 995.9)and number of fecal eggs(3 803.9±110.9)in re-combinant protein group were significantly higher than those of PBS control group (20±3.4, 49 318.1±6 648.3, 5 238.1±303.5, respectively)(P<0.05). There was a high level of specific IgG against rSj29 (maximum dilution 1 ∶ 32 000)in recombinant protein group. Immunohistochemical analysis showed the Sj29 protein expressed on the surface of different stages of S. japonicum. mRNA level of Sj29 was the highest at the 32nd day post-infection. Conclusion The recombinant protein rSj29 induces certain degree of protective immunity in mice.
    Cloning, Expression and Immunogenicity Analysis of Cysteine Proteinase 3 of Trichomonas vaginalis
    JIAWan-zhong;LIZhi;ZHAOLiang;NIEFang-fang;LUNZhao-rong*
    2009, 27(6):  7-487. 
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    Objective To study the humoral antibody response of mice to the recombinant Trichomonas vaginalis cysteine proteinase(TvCP3)in order to investigate the function of the proteinase and its application in diagnosis. Methods T. vaginalis cysteine proteinase gene 3(TvCP3)was cloned by using PCR, and was inserted into the expression vector pET28b. The recombinant plasmid pET28b-TvCP3 or pET28b-TvCP3C(a matured and pre-matured enzyme fragment)was then transformed to Escherichia coli BL21(DE3). The recombinant protein was expressed in E. coli,purified by IMAC(immobilized metal affinity chromatography), and was used to immune BALB/c mice. The mice were divided into groups TvCP3, TvCP3C and control, 6 in each group. The first and second injections for each mouse were administered with 25 μg purified TvCPs which was emulsified with an equal volume of Freund’s complete and incomplete adjuvants, respectively. Two more injections were done using 12.5 μg purified antigens without adjuvants, with 2 weeks interval between the first three injections and one week interval between the 3rd and 4th injections. The murine serum samples were detected one week post the 4th injection. The specific IgG antibody in the serum against the recombinant protein was evaluated by ELISA and Western blotting. Results The expression level of TvCP3 and TvCP3C reached to more than 25% of the total amount of proteins expressed by the bacteria respectively, and the purity in both of them was more than 80% after purified by cobalt-based IMAC resin. ELISA showed that both purified recombinant TvCP3 and TvCP3C induced a high titer of antibodies in the immunized mice(1/204 800 and 1/102 400, respectively). Western blotting analysis indicated that the antibodies reacted with a specific band of the whole T. vaginalis antigens or soluble fractions from T. vaginalis cultures. Conclusion The recombinant TvCP3 and TvCP3C proteins have been highly expressed in E. coli BL21(DE3)and the expressed products can induce high level of antibodies in BALB/c mice, which recognized a specific band of proteins from T. vaginalis soluble fractions and cultures in vitro.
    Recombinant Plasmodium yoelii Expressing Green Fluorescent Protein in Erythrocytic and Mosquito Stages
    FUYong;DINGYan;ZHOUTao-li;CHENJi-de;PENGXiao-hong;XUWen-yue*
    2009, 27(6):  8-491. 
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    Objective To generate recombinant Plasmodium yoelii BY265 strain which can express green fluorescent protein (GFP) in erythrocytic and mosquito stages. Methods Recombinant plasmid containing P. berghei ssu-rrna and GFP genes were linearized by SacⅡ. The linearized plasmid was introduced into the erythrocytic stage of P. yoelii by electroporation. Kunming mice were infected with the recombinant parasites. After 24-30 hours post-infection, mice were treated with 70 μg/ml of pyrimethamine intraperitoneally for 5-6 d, and tail vein blood was then collected to make smear for parasite count. The parasites were examined by PCR. Anopheles stephensi mosquitoes were used to bite mice infected with the recombinant parasite. At the 7th and 16th day after the bite, oocyst development in mosquitoes was observed by fluorescence microscopy. Results The recombinant parasites expressed GFP in erythrocytic stage, and the GFP and ssu-rrna genes were amplified by PCR in the recombinant parasites. The mosquito infection experiment showed a normal development of the recombinant parasites. Conclusion Transgenic P. yoelii BY265 strain has been established with stable expression of GFP in both erythrocytic and mosquito stages.
    实验研究
    Immunogenicity of p55 Gene Fragment from Pneumocystis carinii
    CHENJin-ling;DUANYi-nong*;WANGJian-xin;ZHUDan-dan;QINYong-wei
    2009, 27(6):  9-497. 
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    Objective To evaluate immunogenicity of the recombinant protein GST-p55/570 of Pneumocystis carinii. Methods The fusion protein GST-p55/570 was expressed from the prokaryotic expression plasmid pGEX-570, and purified by using glutathione-agarose. The expressed product was analyzed by SDS-PAGE. Thirty-three mice were randomly divided into three groups, immunized with GST-p55/570, GST and PBS, respectively. Each group was immunized for four times at 2 week intervals. At the 7th day after final immunization, spleen was removed to obtain single cell suspension. Proliferation ability of lymphocytes was determined by MTT. Serum samples were collected at pre-immunizaton and two weeks after each immunization. Antibody level in sera of mice was determined by ELISA. The immune response to the recombinant GST-p55/570 recognized by sera of immunized mice was examined by Western blotting. Results The expressed fusion protein GST-p55/570 showed a Mr 47 000. Compared with GST group (1.134 5±0.073 5) or PBS group (1.124 8±0.041 6), a higher stimulation index (2.063 0±0.160 2) was revealed in GST-p55/570-immunized mice(P<0.01). At the 14th to 49th day after immunization, the antibody titer against GST-p55/570 in the immunized group was significantly higher than that of GST or PBS groups (P<0.01). Western blotting indicated that the fusion protein (GST-p55/570) had specific immune response to positive serum. Conclusion The fusion protein GST-p55/570 elicits significant humoral and cellular immune responses.
    Field Evaluation of Gold-Immunochromatographic Assay for Diagnosis of Vivax Malaria
    WANGJun-yun;WANGJian-jun;SHIFeng;XUXian;YANGYue-tao;GAOChun-hua;ZHENGXiang;GEJun;TANGLin-hua
    2009, 27(6):  10-502. 
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    Objective To evaluate a gold-immunochromatographic assay(GICA) for malaria diagnosis in an endemic area of vivax malaria. Methods Blood samples were collected from febrile patients in 5 township-hospitals of Mengcheng County, Anhui Province, between September and October 2008. The samples were examined by GICA and microscopy under double blind condition and the results were compared. Results Among 292 blood samples, 181 were found P. vivax-positive by microscopy, and 163 were positive by GICA. Altogether, the coincidence of the two methods stood for 92.8%(271/292), including 108 negatives and 163 positives. 21 samples with discrepancy covered 18 microscopy positive but GICA negative, 3 microscopy negative but GICA positive. The GICA positive rate in patients with a parasitaemia of >1 000 parasites/μl,100-1 000 parasites/μl,and <100 parasites/μl was 93.5%(115/123)、86.0%(43/50), and 62.5%(5/8), respectively. Conclusion GICA is a useful diagnosis method for endemic area of vivax malaria.
    Isolation of Microsatellite DNA and the Polymorphic Locus Screeningfrom Phlebotomus chinensis (Diptera ∶ Psychodidae)
    ZHANGLi;FANYong;MAYa-jun*
    2009, 27(6):  11-507. 
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    Objective To isolate the microsatellite DNA of Phlebotomus chinensis and screen the polymorphic loci. Methods Genomic DNA fragments were hybridized with biotinylated oligonucleotide probes(AAT)17,(GA)25, (CCT)17,and(TG)18. The hybridized fragments were captured with Vectrex Avidin D and enriched by centrifugal ultrafiltration with ultra-4-column ultrafiltrate. The target fragments were amplified, cloned and sequenced. The suitable microsatellite loci were chosen in Ph. chinensis’s library to establish PCR amplification assay. The polymorphism screening was conducted by PAGE with Ph. chinensis field specimens. Results The enrichment protocol used in this study was efficient, with a percentage of recombinant clone as 78.6%. There were 118 microsatellite sequences in library, the GenBank accession numbers were from FJ919812 to FJ919932(except GenBank accession numbers FJ919833, FJ919836,and FJ919869). There were 72 typical microsatellite sequences occupying 61.0% and the rest were 46 nontypical microsatellite sequences in the library. Twenty-two loci were chosen to polymorphism screening and PAGE showed that 14 loci were polymorphic. The loci of dinucleotide repeat were more polymorphic than those of trinucleotide and polynucleotide repeat. Conclusion The microsatellite-containing library of Ph. chinensis has been constructed with 118 sequences, and 14 new polymorphic microsatellite loci are reported.
    现场研究
    The First National Survey on Natural Nidi of Angiostrongylus cantonensis in China
    ZHANGYi;LVShan;YANGKun;LIUHe-xiang;HULing;LILi-sha;DENGZhuo-hui;ZHANGHong-man;HUXi-min;YAOLi-nong;ZENGXiao-jun;LIZheng-xiang;CHENZhao0;WANGLi-ying0;ZHOUXiao-nong*
    2009, 27(6):  12-512. 
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    Objective To reveal the natural distribution of Angiostrongylus cantonensis in the mainland of China. Methods The potential areas of A. cantonensis and its main intermediate host Pomacea canaliculata were predicted based on degree-day models using GIS technique. A grid sampling was performed on the prediction map and 5% grids were randomly sampled. A total of 55 sampled sites were selected for the survey on A. cantonensis and its hosts in September-October of 2006. Results Nineteen Provinces/Municipalities/Autonomous Regions were found as potential habitats for P. canaliculata in the mainland of China. It was then confirmed that the snails distributed in the provinces of Fujian, Jiangxi, Zhejiang, Hunan, Guangdong, Guangxi, Hainan, and Yunnan. Higher prevalence of A. cantonensis in P. canaliculata was detected in Jianou of Fujian (36.6%), Xingguo of Jiangxi (19.9%), Rui′an of Zhejiang (16.0%), Rucheng of Hunan (5.0%), Huazhou of Guangdong(6.3%), Shangsi of Guangxi (39.1%) and Wuzhishan of Hainan (25.0%). Conclusion Natural nidi of A. cantonensis have been found in seven provinces where natural infection in P. canaliculata has been detected.
    综述
    Research Progress on the Efficacy, Metabolism and Bioavailability of Mebendazole in Hydatid Disease
    LIUCong-shan;ZHANGHao-bing*
    2009, 27(6):  13-519. 
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    Mebendazole is currently used in the treatment of hydatid disease. Its poor absorption from the gastrointestinal tract and low bioavailability aroused further research on new formulations of mebendazole to increase the bioavailability and improve the therapeutic efficacy. This review summarizes the recent research progress.
    Research Progress on Molecular Identification and Biologic Behavior of Taenia saginata in Western China
    BAOHuai-en*;MOURong
    2009, 27(6):  14-526. 
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    This article reviews the epidemiological investigation of taeniasis saginata in 10 counties of 7 provinces/autonomous regions in western China. The morphological observation of adult worms, molecular identification of 10 isolates of the worms, experimental infection on pigs and cattle with Taenia saginata and T. asiatica, observation on development and biology behavior of cysticercus, and pathological changes in the intermediate host pig and cattle revealed that T. asiatica is a new species, instead of a subspecies of T. saginata.
    Chicken Egg Yolk Immunoglobulin and its Application in Medicine
    CAIYu-chun;CHENJia-xu*
    2009, 27(6):  15-530. 
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    Egg yolk immunoglobulin(IgY)is a major immunoglobulin from birds, amphibians and reptiles. IgY does not activate mammalian complement system, and does not bind to mammalian rheumatoid factors, protein A or G. In addition, it has a high yield, and is easy to extract and purify. With these advantages,IgY has been widely used in veterinary science, functional food and bio-products, and proved potential application for disease control and treatment. This paper reviews biological characteristics of IgY, its extraction, purification, and application in immunodiagnosis and treatment.
    研究简报
    Acaricidal Activity of Clove Bud Oil against Dermatophagoides farinae (Acari ∶ Pyroglyphidae)
    LIJing;WUHai-qiang;LIUZhi-gang*
    2009, 27(6):  16-493,. 
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    Volatile oil from the clove bud was extracted by petroleum ether using Soxhlet Extractor. The acaricidal activity was examined using direct contact and vapour phase toxicity bioassays. In a filter paper contact toxicity bio-assay, at 2.5 h after treatment, clove bud oil at a dose of 12.20 μg/cm2 killed all dust mites. As judged by 24-h LD50 values, potent fumigant action was observed with clove bud oil (12.20 μg/cm2), showing an adequate acaricidal activity against indoor Dermatophagoides farinae.
    Retrospective Analysis of Renal Echinococcosis Confirmed by Surgery and Pathology: 3 Cases
    TIANXiao;YINXiao-ping;ZHOUHuan;LIANGGuang-lu
    2009, 27(6):  17-499,. 
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    A retrospective study was carried out to analyze 3 cases of renal echinococcosis confirmed by surgery and pathology in the hospital during 1999-2008. The patients were examined by 64-CT plain scan and multiple period enhancement scan. Among them, 2 were multi-daughter-cyst hydatid disease, and 1 appeared as solitary one. Two cases were once misdiagnosed as renal cyst and renal hamartoma, and the other was diagnosed correctly before operation. CT signscanning combined with epidemiological information can make an accurate diagnosis for renal echinococcosis.
    Prokaryotic Expression of Bm86 Gene of Boophilus microplus and Optimization of the Expression Condition
    MAMi-ling;GUANGui-quan;LIYou-quan;LIUAi-hong;RENQiao-yun;NIUQing-li;YINHong;LUOJian-xun*
    2009, 27(6):  18-533. 
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    A pair of specific primers was designed based on the reported Bm86 gene of Boophilus microplus,the Bm86 gene was cloned by PCR using the plasmid pMD18-T-Bm86 as templates,and subcloned into the prokaryotic plasmid pGEX-4T-1. The recombined plasmid was transformed into E. coli BL21(DE3) and followed by expression of the protein induced by different concentration of IPTG for different time. SDS-PAGE showed that the recombinant plasmid pGEX-4T-1/Bm86 expressed a fusion protein Bm86-GST (Mr 94 000) after being induced with IPTG. High level expression of Bm86-GST was found at 1 mmol/L IPTG condition after incubation for 8 h at 37 ℃, and the expression level of the recombinant Bm86-GST reached up to 29% of total E. coli proteins. Western blotting analysis showed that the recombinant Bm86-GST was recognized by the rabbit anti-B. microplus positive serum.
    Construction of DNA Vaccine pcDNA3.1(+)/Tetraspanin 2-A against Schistosoma japonicum and its Immuno-protective Effect in Mice
    ZHANGPeng;ZHANGWei-na;RENCui-ping;LIUMiao;SHENJi-jia*
    2009, 27(6):  19-536. 
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    Tetraspanin 2-A (SjTsp2-A) gene was amplified by PCR. pcDNA3.1(+)/SjTsp2-A recombinant plasmids were constructed and transformed into E. coli DH5α. Twenty four BALB/c mice were randomly divided into pcDNA3.1(+)/SjTsp2-A group (A), pcDNA3.1(+)/SjGST group (B) and pcDNA3.1(+) group (C). Each mouse was injected through musculus quadriceps femoris by three times (two weeks interval) respectively with 100 μg pcDNA3.1(+)/SjTsp2-A, pcDNA3.1(+)/SjGST, or pcDNA3.1(+). At two weeks after the final inoculation, mice were each challenged by 40±2 cercariae of S. japonicum. Forty-five days after infection, all mice were sacrificed, the number of worms collected and eggs in liver tisssue was counted. Anti-pcDNA3.1(+)/SjTsp2-A antibody was detected by ELISA and protein expression in quadriceps muscle by immunohistochemical staining. The worm reduction rate (44.4%) and egg reduction rate (28.4%) of group A was higher than those of group B and C (P<0.05), but no significant difference between groups B (3.9%, 19.3%) and C. Higher antibody titer (1 ∶ 25 600) was detected in sera of group A. Immunohistochemistry analysis showed an expression of specific antigens in quadriceps muscles of groups A and B. The DNA candidate vaccine induces partial protective immunity against S. japonicum in BALB/c mice.
    Seroepidemiological Survey on Echinococcosis in Primary School Pupils of Liyang City of Jiangsu Province
    HUANGLi-zhong
    2009, 27(6):  20-537. 
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    Three towns with reported echinococcosis cases and five randomly selected towns were chosen and Echinococcus infection in pupils (7-12 years old) was investigated serologically in 2008. The sero-positive rate (IgG) was 0.9%(25/2 768). There was no significant statistical difference on the IgG positive rate between rural and urban areas (χ2=2.82, P>0.05), males and females (χ2=0.32, P>0.05).
    病例报告
    An Indigenous Case of Plasmodium malariae Infection in Tengchong County of Yunnan Province
    WANGJia-zhi;LIUHui;LIXin-he;FANDa-hong;YINXue-mei
    2009, 27(6):  21-482. 
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