Table of Content

30 April 2009, Volume 27 Issue 2

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论著

Tissular and Subcellular Localization of the ATP Synthase B Subunit in Clonorchis sinensis

HUXu-chu;ZHOUHong-juan;HUFeng-yu;MAChang-ling;ZHAOJun-hong;HUANGCan;ZHENGXiao-ling;XUJin;YUXin-bing*

2009, 27(2):  1-101. 

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Objective To illustrate the distribution of ATP synthase b subunit in the tissue of Clonorchis sinensis adult and its subcellular mimical localization in HeLa cells. Methods With the antiserum against recombinant CsATP-synt_B protein raised from SD rats as primary antibody， paraffin sections of the adult of C. sinensis were processed by the method of fluorescent immunohistochemistry to observe the distribution of CsATP-synt_B protein in adult worm. According to the prediction by bioinformatics of the definite mitochondrial targeting sequence（MTS）and probable Bipartite nuclear localization signals（NLS_BP）in CsATP-synt_B sequence， recombinant pEGFP-N1 plasmids containing the intact and three defective CsATP-synt_B sequence with single defect of MTS or NLS_BP or double defect respectively were constructed. The recombinant plasmids and the control plasmid-pEGFP-N1， pEYFP-Mito and H2B-CFP， were transfected into the HeLa cells by LipofectamineTM 2000 reagent and the subcellular location of the GFP fusion protein was observed with confocal microscopy. Results The CsATP-synt_B protein appeared to distribute all over the adult worm， especially abundant on the acetabulum， ovary， vitellarium and tegument. The intact CsATP-synt_B was definitely expressed in mitochondria and/or nucleus of infected HeLa cells， whereas the MTS-deleted mutant only in cytoplasma and nucleus， the NLS_BP-deleted mutant in mitochondria and cytoplasm， and the double defect mutant only in cytoplasm. Conclusion The distribution of CsATP-synt_B in adult is accord with that of mitochondria， and mainly exits in the organsand the tissues of active energy metabolism. This study first predicted and confirmed that CsATP-synt_B can be expressed in the nucleus.


Comparative Proteomic Analysis of the Promastigotes and Amastigotes of Leishmania donovani

JINGBao-qian*;DENGShi-shan;ZHANGRen-gang;ZHANGJie

2009, 27(2):  2-106. 

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Objective To explore the protein profile and identify developmentally regulated proteins of the promastigotes and axenic amastigotes with comparative proteomics technique. Methods The total proteins of promas-tigotes and axenic amastigotes of Leishmania donovani SC6 strain were separated by two-dimensional electrophoresis（2-DE） in a broad pH range（3－10），and the gel was stained with Coomassie blue. The images were analyzed by PDQuest 1.0 software，and the major developmentally regulated proteins were identified by electrospray mass spectrometry. Results Approximately 700 protein spots were revealed in equivalent proteins of the promastigotes and axenic amastigotes separated by 2-DE，among which more than 90% protein spots showed equivalent quantity and distribution，with 6 proteins up-regulated and 3 proteins down-regulated in axenic amastigotes compared with promastigotes. Five of the 6 up-regulated proteins were with known function，respectively ascribed as Reiske iron-sulfur protein precursor，α-tubulin，peroxidoxin 1，dihydrolipoamide acetyltransferase precursor，and mannose-1-phosphate guanyltransferase. Two of the 3 down-regulated proteins were identified as heat shock protein 70 and β-tubulin. The functions of the developmentally regulated proteins were related to the carbohydrate/energy metabolism，stress response，or formation of cell membrane/cytoskeleton. Conclusion The findings demonstrate the differences in protein expression profiles between promastigotes and amastigotes.


Immunoreactivity of the Recombinant Protein of Echinococcus granulosus Antigen B

LVGuo-dong;LIUTao;LINRen-yong;WANGXing;WANGJun-hua;RENZhi-hui;WENHao;LUXiao-mei*

2009, 27(2):  3-110. 

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Objective To express the recombinant antigen B（rAgB）of Echinococcus granulosus（Eg）and investigate its immunoreactivity. Methods The rAgB gene fragments were inserted into pET41a（+）prokaryotic vector. The recombinant plasmid was transformed into E. coli BL21（DE3）and followed by expression of the protein induced by isopropyl β-D-1-thiogalactopyranoside（IPTG）. The protein was purified with sepharose 4B by affinity chromatography, and tested by SDS-PAGE electrophoresis. Its immunoreactivity was examined by Western blotting, and a rapid diagnosis kit for human echinococcosis was used as control. Results The constructed recombinant plasmid pET41a-rAgB was identified by PCR, digestion with restriction enzyme and sequencing. The recombinant rAgB-GST was about Mr 40 800 with a purity of 78.4%. Western blotting showed that the positive rate of rAgB-GST reacting with sera of cystic echinococcosis（CE）, alveolar echinococcosis（AE）, paragonimiasis westermani and clonorchiasis sinensis patients, and healthy persons is 79.2%（95/120）, 51.1%（23/45）, 0（0/32）, 0（0/20）, and 0（0/24）, respectively. Its overall sensitivity and specificity were 79.2%（95/120） and 81.0%（98/121）, respectively, slightly higher than the sensitivity（72.8%, 75/103） and specificity （76.9%, 30/39）of the rapid diagnosis kit for human echinococcosis. Conclusion The rAgB-GST recombinant protein is recognized by the sera of CE and AE patients, showing a proper immunoreactivity.


In Vitro Effect of Secnidazole Benzoate on Trichomonas vaginalis

ZHANGXiao-xiang*;ZHENGShan-shan;ZHUMei-mei

2009, 27(2):  4-114. 

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Objective To research the trichomonacidal effect of secnidazole benzoate in vitro. Methods Trichomonas vaginalis was cultured in liver extract medium in 96-well microplate. The culture suspension of Trichomonas vaginalis was divided into four groups: secnidazole benzoate, secnidazole, metronidazole and control, with medium as blank control. MTT colorimetric assay was applied to determine the inhibitory effect of secnidazole benzoate on the proliferation of Trichomonas vaginalis. The culture suspension was transfered into test tubes and divided into same groups to observe inhibitory effect by the classical microscopic counting method. Results After 24 h incubation, the proliferation of the parasites was concentration-dependent by secnidazole benzoate（t=9.02, P<0.01）at the concentration ranges from 0.15 μg/ml to 20.0 μg/ml with a relative inhibition rate（%）of 14.6，28.7，31.3，60.4，89.0，89.2，95.6，and 100.0 for MTT colorimetric assay, and 18.2，31.1，39.7，68.8，84.6，90.1，94.6，and 100.0 for counting method，respectively. In the period of 6-24 h incubation, the inhibition was in a time-dependent manner. The minimum sterilizing concentration and the minimum inhibitory concentration were 20 μg/ml and 0.15 μg/ml respectively. Conclusion Secnidazole benzoate shows a similar trichomonacidal effect to metronidazole and secnidazole.


Experimental Study on Recombinant Bla g 2（rBla g 2） in the Treatment of Allergic Asthma in Mice

SHENXiao-ying;ZHUQing-xian;LIUZhi-gang*;LIXiang-hui

2009, 27(2):  5-119. 

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Objective To study the therapeutic effect on murine allergic asthma with recombinant Bla g 2 （rBla g 2）allergen and its possible mechanism. Methods Eighteen BALB/C mice were randomly divided into three groups: normal control group（group A）, asthma model group（group B）, and recombinant protein rBla g 2 treatment group（group C）. Mice in groups B and C were subcutaneously immunized weekly with rBla g 2（50 mg）formulated in Al（OH）3 adjuvant for three weeks. Group A received only adjuvant emulsified with normal saline. Two weeks after the last inoculation, mice in group C were administered each with rBla g 2（100 mg）/dose, and groups A and B were given PBS. All the mice received eight doses at 2-day intervals. One week after the last immunotherapy, mice in groups B and C were intranasally challenged with 50 mg rBla g 2 daily for seven days, while mice in group A received PBS. Twenty-four hours after the challenge, the following items were examined: airway hyperresponsiveness of mice, total cellular score and cell classification in bronchoalveolar lavage fluid（BALF）, level of rBla g 2-specific IgE and IgG2a in serum, lung inflammation by HE stain, and Bcl-2 expression of eosinophils of lung by immunohistochemistry. Results Compared with group B, group C showed a decreased Penh value of airway hyperresponsiveness（P＜0.05）, reduced serum rBla g 2-specific IgE but increased IgG2a（P＜0.01）, and reduced Bcl-2 expression of eosinophils. Total cells［（24.60±15.08）×105/ml］and eosinophils［（22.20±3.76）×105/ml］ in BALF of group B significantly increased than those of group C［（14.30±4.95）×105/ml and（5.20±1.56）×105/ml，respectively］（P＜0.01）. The interstitial space surrounding the airway lumen was characterized by a denseｌｙ mixed cellular infiltrate, tissue edema and epithelium tissue damage in group B, while lung inflamma-tion of group C reduced considerably. Each test value of group C was substantially similar to that of group A. Conclu-sion The experiment shows proper immunotherapeutic efficacy of rBla g 2 in murine allergic asthma, which may possi-bly related to the apoptosis of eosinophils.

Conditional Inhibition of Hemozoin Formation by Chloroquine in vitro

SUNJun*;CHENBo;LONGYan-yu

2009, 27(2):  6-124. 

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Objective To study the characteristics of inhibition on hemozoin formation by chloroquine under in vitro condition. Methods Under different concentrations（0.5-2 mol/L）of sodium acetate（NaAc）and at the pH range of 4.0-5.0, chloroquine was tested for inhibition of β-hematin（hemozion）formation by using the HPIA（heme polymerization inhibitory activity）assay. The morphology of β-hematin crystals was determined by light microscopy. Ultraviolet spectrophotometry was employed to measure β-hematin content, and the size of β-hematin crystal was analyzed by X-ray diffraction（XRD）. Results Chloroquine exhibited varied effect on β-hematin formation, depending on pH value and Na+ concentration. When the NaAc concentration increased from 0.5 mol/L （pH 4.2）to 2 mol/L（pH 4.8）, the chloroquine inhibitory effect also increased. Results suggested that there exists a threshold pH, below which the β-hematin formation escalates and chloroquine inhibition declines, and at or above which chloroquine exerts a stronger inhibitory effect on β-hematin formation. With the increase of pH from 4.4 to 4.8, the crystallinity and the size of crystal changed from 6.93% and 357 Å to 6.32% and 264Å, respectively. When pH reached to 5, no more β-hematin formed. Chloroquine could reduce the crystallinity and crystal size of β-hematin at same pH value. Morphology analysis on the samples was consistent with the above results. Conclusion Chloroquine inhibits hemozoin formation only when the pH value is at or above threshold pH.


Establishment and Application of School-based Health Promotion and Intervention Model of Schistosomiasis in Lake-type Endemic Area

CHENHai-ying;HUGuang-han;*;SONGKuang-yu;XIONGZhi-wei;HUJia;YANGPing-yi;PENGGuo-hua;HUWei-chen;YUSong-shan;FUGuo-lan;LIUZhou-hua;QIJing-cheng;GEJun;WANBao-ping

2009, 27(2):  7-129. 

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Objective To establish an intervention model of school health promotion, and apply it in developing “schistosomiasis-free schools”. Methods At the pilot stage, all students of Henghu primary school and Banshan primary school in Xinjian County of Jiangxi Province were selected as experiment group and control group, respectively. A baseline survey covered knowledge and attitude on schistosomiasis control, water contact behaviors and Schistosoma japonium infection rate. Two health promotion intervention models, i.e. “information communication + training of protection skill + reward & punishment” （model A, 1993-1999） and “information communication + behavior participation + encouragement” （model B, 2000-2007）, were implemented in Henghu school. The effect of two models was compared by infection rate. At the application stage, all students of 8 schools in Xinjian County，Nanchang County，and Jinxian County were chosen for evaluation of the effectiveness of Model B with same methods and index. Results Before intervention there was no significant statistical difference on the passed rate of anti-schistosomiasis knowledge, correct rate of anti-schistoso-miasis attitude, frequency of infested water exposure and the infection rate between Henghu and Banshan schools（P＞0.05）. In Henghu school, the intervention showed significant effect on the scores of knowledge and attitude after one year （P＜0.01）, raised from 9.0% and 55.1% before intervention to 94.4% and 98.9% after intervention, respectively. The frequency of infested water exposure and the infection rate significantly decreased from 14.6% and 13.5% before intervention to 1.9% and 2.3%, respectively （P＜0.01）. In 2-7 years after intervention, there were only one or two schistosomiasis cases each year. At the application stage, no schistosomasis cases were found among Model B target population in two successive years after intervention. Conclusion The practice of Model B can be extended to other schools in endemic area to develop “schistosomiasis-free schools”.

Real-time PCR in Analyzing DNA Extraction from Cryptosporidium Oocysts

CHENSheng-xia;WULiang;SHENYu-juan;ZHANGQiu-xia;LITing-ting;JIANGXu-gan;XUYu-xin;CAOJian-ping*

2009, 27(2):  8-134. 

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Objective To compare the quality and quantity of DNA extracted from Cryptosporidium oocysts by different methods. Methods Cryptospordium oocysts were treated with different kinds of lysis buffers from USA Promega（Promega）and Shanghai Generay（Generay）commercial DNA extraction kits，2% Triton X-100 and 5% guanidine thiocyanate. The oocysts were then broken down by freeze-thawing，proteinase K and sonication. Genomic DNA was purified using the commercial kits or Chelex-100. Real-time PCR technique was used to determine the copies of Cryptosporidium oocyst wall protein（COWP）gene. The Promega commercial DNA extraction kit was used as control. Results The Promega kit resulted in a higher copy number of COWP gene[（6.45-9.86）×106]than that of Generay commercial DNA kit [（2.38-3.69）×106 ], 5% guanidine thiocyanate[（1.27-21.29）×105] or 2% Triton X-100 [（2.06-866.70）×103], respectively. The method of freeze-thawing plus proteinase K plus sonication provided the highest copy number of COWP gene. Conclusion The method of freeze-thawing + proteinase K + sonication is most effective. The effect of DNA extraction by Generay kit and 5% guanidine thiocyanate is similar to that of Promega kit.


实验研究

Identification of Calf-origin Cryptosporidium bovis Shanghai Isolate by Nested PCR

YUANZhong-ying;SHENYu-juan*;CAOJian-ping;LIUHui;CHENSheng-xia

2009, 27(2):  9-139. 

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Objective To identify a strain of Cryptosporidium in the feces of naturally infected calf in Shanghai. Methods Stool sample was examined by modified acid-fast staining. The size and morphology of the oocysts were micros-copically determined. Genomic DNA was extracted from the oocysts isolated from feces of a naturally Cryptosporidium-infected calf. According to the sequence of Cryptosporidium 18S rRNA gene, two pairs of primers were designed and syn-thesized. The PCR products was amplified by nested PCR and sequenced in double directions. Homology searches were done over the Web using the program Blast. Phylogenetic tree was constructed with NJ method by MEGA4.0 software. Results Oocysts of the Shanghai isolate were round or elliptical with a size of （5.6±0.49） μm ×（5.2±0.51） μm. Nested PCR resulted in fragments of approximately 810 bp, and the 18S rRNA nucleotide sequence had 100% identity with C. bovis from Brazil （GenBank Accession No: 151935628）. This isolate was clustered in the same clade with C. bovis from Brazil. It showed an identity of 99% with the sequences of C. bovis from Qinghai Ｐrovince of China, Mongolia, USA, and Tunisia. Conclusion The calf-origin Cryptosporidium derived from Shanghai has been identified as C. bovis.


Preliminary Application of PCR-based Assay for the Detection of Neospora caninum in Bovine Aborted Fetus

ANGChun-ren*;ZHAIYan-qing;ZHAOXing-chun;TANQiu-ju;CHENJia;CHENAi-hua;WANGYu

2009, 27(2):  10-143. 

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Objective To establish a PCR diagnostic method based on Nc-5 gene of Neospora caninum，for being used to detect Neospora in brain tissues of bovine aborted fetus. Methods Specific primers were designed and synthesized based on the reported Nc-5 gene of N. caninum（GenBank Accession No. AY459289）. Using genomic DNA from N.caninum as templates, Nc-5 gene was amplified by PCR. The PCR product was cloned into pMD18-T vector, transformed into Escherichia coli JM109 and then sequenced. To evaluate the specificity of the PCR, genomic DNA of Theileria annulata，Babesia bovis，Toxoplasma gondii，Leishmania donovani and standard strain of N. caninum were used as a template in the PCR. For determining the detection limit of amplification procedure, PCR was run on a dilution series of genomic DNA from N. caninum（1.562 5-200 ng/ml）. Brain tissue samples of 32 aborted fetuses were detected by PCR-based assay, and 23 blood samples from mothers were tested by ELISA. Results The amplified DNA fragment （350 bp）had a high identity of 98% with the Nc-5 gene sequence of N. caninum（GenBank Accession No. AY459289）. The PCR was specific for N. caninum and allowed the detection of 3.125 pg DNA of the parasite, while no amplification occurred with the other four species of protozoa. PCR-based assay and ELISA showed a positive rate of 18.8%（6/32） and 17.4%（4/23）of the samples tested, respectively. Moreover，all the 4 antibody positive samples showed PCR positive. There is no significant difference between the two assays（P>0.05）. Conclusion PCR diagnostic method is promising in detecting Neospora infection in brain tissues of aborted bovine.

Influence of Baicalin on TNF-α and Soluble Intercellular Adhesion Molecule-1 in Rats Infected with Pneumocystis carinii

ZHOULing;ZHOUBi-ying*

2009, 27(2):  11-147. 

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Objective To explore the effect of baicalin on tumour necrosis factor-α （TNF-α） and soluble inter-cellular adhesion molecule-1 （sICAM-1） of immunosuppressed rats infected with Pneumocystis carinii. Methods Forty-nine SD rats were randomly divided into 7 groups: normal control （A）, immunosuppressed control （B）, SMZ/TMP control （C）, baicalin prevention （D）, low dose （E）, moderate dose （F） and high dose （G）. Rats of group D received six injections at three-day intervals with 3.5 mg dexamethasone sodium phosphate for 3 weeks, while groups B, C, E, F and G received same immunosuppressor but for 6 weeks. Rats in group D were given 100 mg/kg baicalin daily for 5 weeks, and groups C, E, F and G were given 250 mg/kg SMZ+50 mg/kg TMP, low dose ［100 mg/（kg?d）］, moderate dose ［200 mg/（kg?d）］, and high dose ［400 mg/（kg?d）］ baicalin daily for 2 weeks, respectively. At the end of 8th week after immunosppression, the contents of TNF-α and sICAM-1 in peripheral blood were detected by radioimmunoassay（RIA） and ELISA, respectively. The pathological change of lung was observed by lung imprint smear with gormori methenamine silver （GMS） staining and lung section with hematoxylin-eosin （HE） staining. Results The content of TNF-α in group D［（2.14±0.14） ng/ml］， group E ［（2.57±0.15） ng/ml］， group F ［（1.46±0.14） ng/ml］, group G ［（1.12±0.13） ng/ml］ andgroup C ［（1.59±0.14） ng/ml］ were higher than that of group A ［（0.70±0.21）ng/ml］（P<0.05, P<0.01）, and lower than group B ［（3.65±0.73） ng/ml］（P<0.01）. The content of sICAM-1 in group D ［（618.68±52.42） pg/ml］， group E ［（814.29±61.11） pg/ml］， group F ［（498.08±32.56） pg/ml］, group G ［（377.06±56.56） pg/ml］ and group C ［（386.95±44.98） pg/ml］ were lower than group B ［（1 247.39±288.57） pg/ml］（P<0.05）. Compared with immunosuppressed control group, there were less alveolar interstitial lymphocytes, foamy intra-alveolar exudate and inflammation of lung tissue in rats of drug treatment groups. Conclusion Baicalin can decrease the contents of TNF-α and sICAM-1， and alleviate inflammation in lung tissues of rats infected with Pneumocystis carinii.

Experimental Establishment of Life Cycle of Clonorchis sinensis

LIANGChi;HUXu-chu;LVZhi-yue;WUZhong-dao;YUXin-bing;*;XUJin;ZHENGHuan-qin;

2009, 27(2):  12-150. 

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Objective To establish and maintain the life cycle of Clonorchis sinensis in laboratory. Methods Adult worms and eggs of Clonorchis sinensis were collected from naturally infected cats. Eggs were ingested by freshwater snails in aquarium. When the cercariae were released from infected snails, they invaded into freshwater fishes. From the 30th day on after the release of cercariae, the infection rate and metacercariae density in freshwater fishes were determined. Results After 95 days the infected snails began shedding cercariae in a temperature range of 24.3 -37.2 ℃, and no cercariae were found under 20 ℃. The infection rate in the snails Parafossarulus striatulus and Alocinma longicornis was 12.5% and 18.0%, respectively. Metacercariae were found in fish at 30 days after cercariae infection, and matured metacercariae were detected in 45 days. The number of metacercariae per gram of fish meat in Pseudorasbora parva, Ctenopharyngodon idellus, Rhodeus sinensis, Hypophthalmichthys nobilis, Cirrhinus molitorella, Carassius auratus, Cyprinus carpio and Oreochromis niloticus was 1 792, 16, 8, 6, 5, 4, 4, and 2, respectively. Rats and cats were fed with metacercariae from fish to receive adult worms. Conclusion Life cycle of Clonorchis sinensis has been established and maintained in the laboratory.


现场研究

Distribution of Food-borne Parasitic Diseases and Dietary Habits in Human Population in Guangxi

LIXue-ming;OUYANGYi;YANGYi-chao;LINRui;XUHong-bo;XIEZu-ying;LIShu-lin;SHANGShao-ming

2009, 27(2):  13-155. 

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Objective To investigate the distribution of common food-borne parasitic diseases and dietary habits in Guangxi. Methods In the survey from 2002 to 2005，questionnairing，fecal examination，ELISA or skin test were applied in the endemic areas of clonorchiasis，taeniasis，cysticercosis，trichinelliasis or paragonimiasis in Guangxi Zhuang Autonomous Region. 13 990 residents at 27 sites in 9 counties were examined for clonorchiasis，22 772 residents at 22 sites in 10 counties for taeniasis，9 638 residents at 20 sites in 10 counties for trichinelliasis and 9 347 residents at 18 sites in 6 counties for paragonimiasis. Results Among 109 counties, residents in 64 counties（59.8%）were found with local habit of eating raw meat, covering Han and other 6 ethnic groups（Zhuang，Yao，Miao，Maonan, Dong and Buyi）. Various animal food such as fish，pork，pork liver，swine blood，beef，ox blood，chicken blood，goat blood，crab，shrimp，clamworm，snail，oyster，and duck meat were consumed in raw with or without seasonings，or by ways of hot-pot，pickle，dipping in boiling water or wine-soaked. Food-borne diseases such as clonorchaisis，taeniasis，cysticercosis，trichinelliasis and paragonimiasis were prevalent in the Region. The survey showed that clonorchiasis is prevalent mainly in the river basins of Yongjiang, Zuojiang and Youjiang in the middle-west and southern part of the Region with a prevalence of 9.8%，teaniasis and cysticercosis in the north and middle-north such as Rongshui prevalence of 43.3% and 2.1%respectively，Bingyang(2.1% and 1.0%) and Tianlin (1.6% and 1.1%)counties. Sporadical cases of trichinelliasis were found in the counties of Debao，Baise，Luzhai and Nandan with positive rate 9.8%，6.4%，5.3% and 3.1% by serological test. Skin test on paragonimiasis among residents in sampled areas revealed a positive rate of 2.6% and the cases mainly distributed in Rongshui, Longsheng and Quanzhou counties. Conclusion Consumption of raw or undercooked meat is popular in the local residents especially the ethnics, which is an important factor in the prevalence of food-borne parasitic diseases.


Effect of Synchronous Chemotherapy for Residents and Livestock against Schistosomiasis Japonica for 12 Years in Jingzhou City

HELiang-cai;ZENGwei;WANGJia-song*;YUANMei-zhi;DONGJuan;FUZheng-yin

2009, 27(2):  14-160. 

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Objective To evaluate the anti-schistosomiasis effect of World Bank loan for schistosomiasis control project 1992-2001 in Jingzhou City, Hubei Province, and observe the endemic situation after two years of the end of the project. Methods By a retrospective longitudinal survey，information from annual reports was collected, which included disease screening, treatment and extended chemotherapy for human population and livestock, the distribution of Oncomelania hupensis and health education information from eight counties in Jinzhou prefecture from 1992 to 2003. Statistical u test was used for phased retrospective dynamic analysis. Results In the year 2001, schistosomiasis patients（77 009 cases）were 45.2% less than that of 1992（140 438 cases）, The number of acute and advanced cases（63 and 1 032 cases, respectively）was 74.2% and 43.9% less than those of 1992（244 and 1 841 cases, respectively）. The average infection rate among residents was also reduced from 7.8% of 1992 to 4.7%（u=28.864, P＜0.01）. Number of infected cattle in 2001（3 454）was 57.0% less than that of 1992（8 035）, and the prevalence in cattle was reduced from 7.9% of 1992 to 3.7%，decreased by 53.2%（u=9.338, P＜0.01）. Snail-ridden area in 2001 was 19 368 hectares, 23.7% less than that of 1992（25 395 ha）. In 2003, two years after end of the project, number of acute schistosome infections was 100 cases, 59.0% less than that of 1992, but 58.7% more than that of 2001. The average infection rate in humans increased to 8.4% in 2003, 7.7% and 78.7% more than that of 1992 and 2001, respectively（u=6.453、34.804, P＜0.01）. The average infection rate in cattle increased to 6.5% in 2003, 17.7% less than that of 1992, but 75.6% more than that of 2001（u=3.061、5.894，P＜0.01）. Snail-ridden areas also increased in 2003 with 29.3% and 69.6% more than that of 1992 and 2001, respectively. Conclusion Synchronous chemotherapy for residents and livestock can reduce the prevalence of schistosomiasis and control its transmission effectively in several year period, but the disease may re-emerge after chemotherapy stopped.

综述

Research Progress on the Technology of Microsporidian Detection

WANGYong-bin;LIUJi-ping*

2009, 27(2):  15-166. 

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The microsporidia are obligate intracellular eukaryotic parasites. They have been paid more attention as being the emerging pathogen of human， so it is important to control microsporidiosis using fast and precise detecting technology. In order to provide a reference for controlling microsporidian infection effectively， this paper reviews the progress of studying on the detecting technology from the microscopic staining methods， immunological and molecular biology.


GIS/RS/GPS Application in the Study of Relationship between Natural Factors and Malaria Transmission

ZHANGShao-sen;ZHOUShui-sen*

2009, 27(2):  16-170. 

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Malaria transmission is highly related to the natural factors such as temperature, rainfall and topography. Recently, the GIS/RS/GPS techniques were developed based on computer science and spatial analysis technology, which were widely used in data collection and analysis, establishment of mathematics model. This paper reviews the applications and development of GIS/RS/GPS techniques in the studies of relationship between natural factors and malaria transmission.


Research Progress on Honeybee Microsporidia

WANGQiang;ZHOUTing*;DAIPing-li;WUYan-yan;LIUFeng

2009, 27(2):  17-174. 

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This paper summarized the new hotspots in honeybee microsporidia, covering the progress on taxonomic status, infection mechanism, diagnosis method, epidemiological regularity and control.


研究简报

Sensitivity of Plasmodium vivax to Chloroquine in Laza City, Myanmar

LIANGGui-liang;SUNXiao-dong;WANGJian;ZHANGZai-xing;*

2009, 27(2):  18-176. 

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Fourty-eight patients infected by Plasmodium vivax were randomly divided into chloroquine regimen groups A and B. Each case in group A received a total dosage of 1 200 mg（base） over a 3-day period （600 mg on the first day then 300 mg daily）. Cases in group B each received total dosage of 1 500 mg （base） in 3 days （750 mg on the first day then 375 mg daily）. Body temperature and blood smears were examined on days 0, 1, 2, 3, 7, 14, 21 and 28, respectively. All patients were negative for asexual parasites within 3 days. The cure rate on day 28 was 100%. Results showed that P. vivax is sensitive to chloroquine in Laza, Myanmar, and chloroquine can still be used for the imported vivax malaria cases at the China-Myanmar border.


Cloning and Sequence Analysis of the egG1Y162 Gene of Echinococcus granulosus

CAOChun-bao;MAXiu-min;DINGJian-bing;*;JIAHai-ying;MAMUTY?Wulamu;MAHai-mei;WENHao

2009, 27(2):  19-179. 

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A pair of primers （egG1Y162） were designed according to the nucleotide sequence of Echinococcus multilocularis emY162 antigen gene. Using genomic DNA and cDNA from protoscoleces and adult worms of E. granulosus as templates, PCR was performed with the primers to obtain fragments of egG1Y162 gene. PUCm-T/egG1Y162 rec-ombinant plasmids and PUCm-T/egY162 cDNA recombinant plasmids were constructed and identified by PCR, digestion with restriction enzyme and sequencing. The egG1Y162 antigen gene was amplified in protoscoleces and adult worms of E. granulosus. The size of the egG1Y162 gene was 1 648 bp and cDNA was 459 bp, and GenBank accession numbers were AB458258 and AB458259, respectively.


Routine Blood Test and Bone Marrow Examination in Visceral Leishmaniasis Patients in Kashi Prefecture

ZHANGHui-qin;YUHong-Bin;WANGHuan

2009, 27(2):  20-181. 

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The examinations of blood and bone marrow in 125 patients with visceral leishmaniasis revealed that seven routine parameters of the blood samples were below the lower limit of normal in most cases and 64.8% of the cases showed complications with iron deficiency anemia （IDA） and cell maturation disorders. Therefore, iron deficiency anemia and significant reduction of blood components can be used as indicators on the severity of Leishmania infection.


Epidemiological Investigation on an Outbreak of Amoebic Dysentery in Jiangshan City of Zhejiang Province

MAOLi-xiong;ZHENGJian-jun*;WANGXin-ming;YAOLi-nong;CHENGHua-liang;ZHUQiu-guo

2009, 27(2):  21-183. 

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The investigation was carried out from 15 May to 10 June 2006 among diarrhea patients of two schools and periphery residents in Hecun Town, Jiangshan City. Stool samples were examined for Entamoeba histolytica. Water samples were taken for microbial analysis. 31 cases with E. histolytica were found，with 74.2% （23 cases） of students and preschool children. 9 cases were found in Liuyi kindergarten and 8 cases in Hecun central primary school, with a prevalence of 7.4% and 0.65%， respectively. Among 594 asymptomatic close contactors, 9 cases （1.5%） were carriers of cysts. Of the 31 cases, 22 were found with no habit of handwashing before eating or after defecation, and 14 cases had a close contact to the patients. No amoebic cysts or trophozoites were found from 12 water samples collected from schools or patient′s houses, but the Esherichia coli level exceeded the national standard in 7 samples.


Morphological Comparison of Two Species of Theileria in Sheep

LIYou-quan;GUANGui-quan;MAMi-ling;LUOJian-xun;YINHong*

2009, 27(2):  22-186. 

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Two splenectomized sheep were infected respectively with Theileria luwenshuni and T. uilenbergi, 2 spec-ies identified by PCR. When piroplasms were found in blood smears from ears of the sheep, morphological observation on the Theileria spp. was carried out by optical microscopy. By Giemsa staining, the cytoplasm exhibited in slight blue and nucleus in purple. The two Theileria species displayed various shapes, but pyriform-shaped, round-shaped and needle-shap-ed parasites appeared in every stage of the infection. In fact, there is no significant morphological difference between the two species.


Field Investigation of Sparganosis in Frogs in Huaxi Area

MAOJia-zhi;ZHOUBi-jun;WANGKai-gong;WENMing*

2009, 27(2):  23-188. 

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Frogs were caught from 4 towns in Huaxi of Guiyang and dissected. The collected spargana were used to infect young dogs for species identification. Results showed that the wild frogs were identified as Rana nigromaculata, and the infection rate was 16% （131/818） with an average intensity infection of 3.44 per frog, The tapeworm obtained from an infected dog was specified as Spirometra mansoni.