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Table of Content

    30 April 2008, Volume 26 Issue 2
    论著
    Effect of Paeoniflorin on Secretion of TGF-β1 from Macrophages in Mice
    CHUDe-yong;LICong-lei;LIJing;LUOFei;ZHENGMei-juan;WUQiang;LUOQing-li;SHENJi-long*
    2008, 26(2):  1-85. 
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    Objective To explore the effect of paeoniflorin (PAE) on the production of transforming growth factor β1 (TGF-β1) from peritoneal macrophages(PMs) stimulated by soluble egg antigen (SEA) of Schistosoma japonicum. Methods SEA was prepared by trituration and added into culture plank, flask and dish containing PMs which were cultured for 24 h. TGF-β1 secreted from PMs was measured by ELISA. TGF-β1 mRNA and protein produced from PMs were evaluated by RT-PCR and Western blotting,respectively. SEA(10 mg/L) 5 ml was added into culture flask and dish containing PMs. PMs were cultured for 12 h, and PAE at different concentrations (0, 7.5, 15, 30, 60, 120 mg/L) was added into the culture flask and dish, and PMs were cultured consecutively for another 12 h and 24 h, respectively. TGF-β1 mRNA and protein from PMs stimulated by SEA were evaluated by RT-PCR and Western blotting, respectively. Results TGF-β1 (235.86±3.43 ng/L) was produced from PMs under stimulation of SEA at 10 mg/L, and the expression of TGF-β1 mRNA and protein in PMs were depressed significantly by PAE in a concentration-dependent manner (r=-0.827, P<0.01; r=-0.952, P<0.01, respectively). Conclusion PAE inhibits the production of TGF-β1 from PMs stimulated by SEA.
    Identification of Six Trichinella Isolates from China by Multiplex PCR
    CUIJing*;ZHAOGui-hua;WANGZhong-quan;JIANGPeng;NIUHong-tao
    2008, 26(2):  2-89. 
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    Objective To identify and classify six isolates of swine-originated Trichinella from China. Methods Five specific pairs of primers were synthesized based on DNA sequence of expansion segment V region and internal transcribed spacers (ITS1 and ITS2) of ribosomal DNA repeat from Trichinella. International reference strains of five Trichinella species [Trichinella spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4) and T. nelsoni(T7)] were used as control. Six swine Trichienlla isolates from Henan, Yunnan, Harbin, Tongjiang of Heilongjiang, Hubei and Tianjin were identified by multiplex PCR and its effecting factors of PCR amplification were observed. Results Electrophoresis results of multiplex PCR products of Trichinella larvae showed that the band (173 bp) of the six isolates was the same as T. spiralis(T1). The specific band (173 bp) was detected by multiplex PCR through amplification from issues of single T. spiralis larva, the larvae conserved in 80% ethanol for 6 months, the larvae stored in 10% formaldehyde, in 0.05% formaldehyde, 0.2% sodium azide or 0.05% merthiotate for 2 weeks,or fresh mouse muscle with larvae. Conclusion All the six swine Trichinella isolates are identified as T. spiralis (T1) by multiplex PCR.
    A Case-Control Study on the Risk Factors ofCerebral Schistosomiasis Japonica
    HUANGYue-e;WANGTian-ping*;ZHANGShi-qing;YAOYing-shui;WUWei-duo;CAOZhi-guo
    2008, 26(2):  3-94. 
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    Objective To explore the risk factors of cerebral schistosomiasis japonica. Methods A total of 37 confirmed cases,diagnosed during 1999-2004 in Anhui Province,were involved in the study as case group. Three controls for each case were selected from the general population either without schistosomiasis,with chronic schistosomiasis or with acute schistosomiasis. The relevant risk factors on the disease were analyzed. Results Four factors with labour intensity, physical, economical and nutritional conditions before illness between case group and normal control group, 7 factors including epilepsy history, hypersensitivity,intensity and times of infection between case group and acute schistosomiasis control group,10 factors including intensity of infection,times of infection and treatment between case group and chronic schistosomiasis control group, all showed statistical significance (p<0.05). Five factors showed statistical significance (p<0.05) between case group and the 3 control groups. Conclusion The intensity of schistosome infection and treatment history are the main risk factors for cerebral schistosomiasis,and the physcial and psychosocial conditions of the patients show some effect on the disease.
    Genotyping and Polymorphism Analysis of Plasmodium falciparumIsolates by Nest-PCR and PCR-RFLP
    ZHANGQing-feng;HUJing-ying;YANGYue-xin;PANWei-qing*
    2008, 26(2):  4-98. 
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    Objective To analyze the efficiency and specificity of MSP2 alleles genotyping for Plasmodium falciparum isolates by Nest-PCR and PCR-RFLP. Methods MSP2 alleles from Plasmodium falciparum isolates of Yunnan and Hainan were genotyped by Nest-PCR and PCR-RFLP, respectively, and the efficiency and specificity of the two methods were analyzed. Results The conventional Nest-PCR method could detect 79.8% (166/208) alleles of MSP2,and 65.7% (65/99) for 3D7 family, but could not identify the type of any allele. While PCR-RFLP showed 25.3% higher genotyping efficiency than Nest-PCR. Moreover, this method could identify the allele types. Conclusion PCR-RFLP genotyping technique is more efficient and specific than conventional Nest-PCR, and it is a convenient tool in the study on molecular epidemiology of malaria.
    Current Prevalence of Clonorchis sinensis Infection in Endemic Areas of China
    FANGYue-yi*;CHENYing-dan;LIXue-ming;WUJun;ZHANGQi-ming;RUANCai-wen
    2008, 26(2):  5-103. 
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    Objective To investigate the current prevalence of Clonorchis sinensis infection in the known endemic areas of China. Methods According to different endemic level and river system, the lamination stochastic group sampling was carried out in 27 Provinces/Municipalities/Autonomous Regions (P/M/A) from 2002 to 2004. Eggs in stool were examined by modified Kato-Katz thick smears method. Results A total of 5 230 cases of Clonorchis infection was found from 217 829 sampled population of 27 P/M/A, and the mean prevalence was 2.40% by which the number of infected people was estimated to be 12.49 millions. The geometry mean worm egg density was 2 208 EPG(stool). Light, moderate and heavy infection occupied 78.93%, 17.40%, and 3.67% respectively. Infection was found in 19 out of 27 P/M/A investigated, with the highest prevalence in the provinces of Guangdong (16.42%, 2 278/13 876), Guangxi (9.75%, 1 365/13 990) and Heilongjiang (4.72%, 636/13 458). Males showed higher prevalence (2.94%, 3 267/111 262) than females (1.84%, 1 963/106 567). There was infection in all age groups,with the highest prevalence in the group of 50 to 59 years(9.16%). By occupations, more infections were found in businessmen (13.42%, 124/924),followed by factory workers ((7.9%, 298/3 773), and retirees and veterans (5.28%, 70/1 327). Adults showed heavier infection in some provinces where the main infection way was raw fish-eating,but in children in other provinces. For geographical distribution, the terrain in network of rivers and lakes showed higher prevalence (5.23%, 687/13 125) than the hilly terrain (2.34%, 1 111/47 552). Prevalence showed family-aggregation feature. Conclusion Clonorchiasis is widely endemic in China with a high prevalence in some provinces. Male adults have been more involved with a family aggregation.

    New Polymorphic Microsatellite Loci Identified Using Genomic Resource for Schistosoma japonicum
    YINMing-bo;ZHUGen-feng;ZHANGXiang-lin;XUBin;MOXiao-jin;SUNSheng-qiang;WANGSheng-yue;DavidBlair;HUWei;FENGZheng*
    2008, 26(2):  6-109. 
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    Objective To identify new microsatellite loci from genome sequence database for the study of polymorphicsm of Schistosoma japonicum. Methods Schistosoma japonicum isolates were obtained from seven endemic sites in China: Tongling and Guichi counties of Anhui Province, Duchang county of Jiangxi Province, Changde and Yueyang Cities of Hunan Province, Shashi City of Hubei Province, Xichang City of Sichuan Province. In order to study the genetic variance, genomic DNAs of 96 individual adult worms were screened against 17 new Schistosoma japonicum microsatellites and the raw data were analyzed by GenMapper 4.0. Furthermore, the varieties of alleles were inverstigated using GenAlEx 6 and genetic distances within a subpopulation (GenClone) and among populations(UPGMA, MEGA 3.1) were analyzed. Results High levels of polymorphism were found between and within population samples, and significant genetic diversity was observed among the seven subpopulations.Within Jiangxi population, most genetic distances (17 loci) among samples range from 25 to 32, indicating a significant genetic diversity. There are three clusters among the seven populations: Jiangxi, Tonglin, Shashi and Changde population, with the genetics distances ranging from 0.017 8 to 0.036 3; Guichi and Yueyang population belong to another cluster, with the genetic distance of 0.024 7; However, Xichang population is an unique group. Its genetic distances to other populations are notable with a range from 0.019 2 to 0.069 3. Conclusion The 17 new polymorphic microsatellites identified may be used as suitable markers for the study on population genetics of Schistosoma japonicum and the genetic variance of the worms seems to be complicated.
    Cloning and Expression of PbTIP Analogous Gene from Plasmodium berghei ANKA and Preparation of its Polyclonal Antibody
    LIYing-hui;ZHAOYa*;LIUZhong-xiang;HUANGYu-xiao;MaoZhang-xiang;XUECai-fang
    2008, 26(2):  7-112. 
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    Objective To clone and express a novel protein analogous to TIP(T cell immunomodulatory protein)of Plasmodium berghei ANKA and prepare its polyclonal antibody. Methods The PbTIP encoding nucleotide sequence was searched from the Plasmodium berghei genomic database and amplified by PCR. The gene was sub-cloned into prokaryotic expression vector pGEX4T-1 and expressed in E.coli BL2l (DE3). After induction by IPTG, the expression of PbTIP-GST fusion protein was characterized by SDS-PAGE and Western blotting. The inclusion bodies of GST-PbTIP fusion protein were injected into BALB/c mouse. Anti-sera were identified by indirect fluorescent antibody test and western blotting. Results The PbTIP-GST fusion protein was successfully expressed in the form of inclusion bodies, by controlling the temperature and concentration of IPTG. Anti-PbTIP-GST sera were acquired with high titer. The sera specifically recognized the PbTIP with a band of 60 000 in P.berghei infected erythrocyte protein. Conclusion PbTIP/GST fusion protein and polyclonal antibody have been obtained.
    实验研究
    Studies on Immunomodulation Effect of Recombinant Sj16 from Schistosoma japonicum on Inflammation Response of Host
    SUNXi;ZHOUHe-jun;LVZhi-yue;ZHANGShuang-min;HUShao-min;ZHENGHuan-qin;FENGMing-zhao;WUZhong-dao;*
    2008, 26(2):  8-118. 
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    Objective To study the immunomodulation effect of the recombinationt Sj16 from Schistosoma japonicum(reSj16) on inflammation response of host. Methods reSj16 expressed in pGEX-4T-1 was purified by GST purification kit and detected by sodium dodecyl sulfate-polyacrylamide gel electropheresis (SDS-PAGE) and mass spectra (MS). The immunomodulation effect of reSj16 was observed by means of dimethylbenzene inducing mouse ear edema model, carrageenan inducing rat voix pedis swell model and acetic acid inducing mouse experiment peritonitis model. Results The soluble protein of reSj16 was obtained and identified by SDS-PAGE and MS. reSj16 1.0,5.0,10.0 μg/kg evidently suppressed the mouse ear edema induced by dimethyl-benzene, significantly mitigated the rat voix pedis swelling induced by carrageenan, and remarkably suppressed the increase of the capillary permeability of abdominal cavity in experiment peritonitis mouse model. Conclusion The results further prove that Sj16 may be a potential immunosuppressive molecule and may have a notable effect on immunomo-dulation.
    Protective Immunity and Antibody Response ofRats Infected with Trichinella spiralis
    Fu-ShiQUAN*;Haeng-SookLEE;Myung-SookCHUNG;Sung-WeonCHO;Kyoung-HwanJOO;Joon-SangLEE;Han-JongRIM
    2008, 26(2):  9-123. 
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    Objective To study the protective immunity and antibody (IgG, IgG1 and IgG2a) response against adult and larva infection of T.spiralis Korean isolate in rats. Methods Forty-six rats were randomly divided into 7 groups. Group A (A1,A2,10 rats) was used for the determination of protective efficacy from adult stage infection, group B (B1,B2, 14 rats) was for the protective efficacy from muscle larva stage infection, group C (C1, C2, 17 rats) was for challenge control, and group D (5 rats) served as normal control. Rats in groups A, B and C were infected with 1 000 T.spiralis muscle larvae, and the infected rats were treated with flubendazole (20 mg/kg, 10 d) at day 7 (A1, A2) and at day 30 (B1, B2). Rats in groups A and B were re-infected with 500 T.spiralis muscle larvae at day 10 after treatment. Rats in groups A1 and B1 were killed at day 7 and day 30 to inspect the reduction of adult worms in the intestines. Rats in groups A2 and B2 were killed at day 30 to detect the reduction of muscle larvae in diaphragms. Rats in groups C and D were killed at the same time, and all rats were bled at the same time. Specific anti-Trichinella IgG, IgG1 and IgG2a were detected by ELISA. Results Adult stage infection induced protective efficacy by 100% against adult stage and by 99.96% against larva stage. Larval stage infection induced protective efficacy by 99.92% against adult stage and 99.89 % against muscle larvae. Anti-muscle stage larval ES Ag (IgG 3.0, IgG1 2.2, IgG2a 0.8) and anti-adult crude Ag antibodies (IgG 1.9, IgG1 0.8, IgG2a 0.3) significantly increased in the muscle larval stage infection compared to normal control (IgG 0.5, IgG1 0.1, IgG2a 0.1) and adult stage infection (IgG 0.5, IgG1 0.09, IgG2a 0.09) (P<0.01). Higher specific IgG1 antibody (IgG1 2.2) in larva stage infection was shown than specific IgG2a antibody response(IgG2a 0.8)(P<0.01). Conclusion Protective immunity against both adult and larva worms has been induced from adult and muscle larva stage infections of T.spiralis.
    Construction and Expression of Recombinant Baculovirus with Schistosoma japonicum SjPP Gene
    YAOLi-xiao;TAOLi-hong;YANGGuan-zhen;WUXiang-fu;CAIYou-min;LINJiao-jiao*
    2008, 26(2):  10-127. 
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    Objective To express the soluble recombinant Schistosoma japonicum SjPP proteins in TN5B1-4 cells. Methods The total RNA was extracted from adult worms of Schistosoma japonicum. The whole coding sequence of SjPP gene was synthesized by RT-PCR and cloned into donor plasmid. The recombinant donor pFastBac-SjPP was transformed into E.coli DH10Bac forming Bacmid-SjPP which was transfected into insect cell with cational lipofectin. The fusion protein SjPP was analyzed with SDS-PAGE and Western blotting. Results The infective recombinant baculovirus Bacmid-SjPP was obtained and SjPP protein was expressed in insect cells. Conclusion The recombinant protein SjPP has been expressed in insect TN5B1-4 cells with proper antigenicity.
    现场研究
    A Newly Identified Endemic Area of Visceral Leishmaniasis in Minfeng County of South Xinjiang Ⅰ. Epidemiological Survey
    YisilayinOSMAN;JINChang-fa;ZUOXin-ping;GUDeng-an;LANQin-xian;TONGSu-xiang;LIXiong;ZHANGYi;RuziguliJUMAHUN;NIUXin-ling;Abulimiti;Muhetaer;Abuduwaili;Kuerban
    2008, 26(2):  11-131. 
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    Objective To investigate the epidemiological status of visceral leishmaniasis in Minfeng county, a newly identified endemic area in south Xinjiang, China. Methods Based on a hint of possible existence of patients, a retrospective survey was carried out house by house in Andier Township of the county to find cases with suspected signs/symptoms of the disease in the past 20 years including those died. Meanwhile, a survey on current status was conducted, including physical examination(liver and spleen palpation) to those under 15 years-old,leishmanin skin test and rk39 immunochromatographic strip test for part of the residents. Results The investigation covered 313 local residents and revealed no case with present illness but 60 cases with a history of suspected signs/symptoms including 13 deaths, traced back as visceral leishmaniasis. Leishmanin skin test was performed in 171 people with a positive rate of 99.4% and 28 out of 29 suspected cases showed positive (96.6%). rk39 immunochromatographic strip test was conducted in 197 people with a positive rate of 10.2% (20/197) and 19.4% (6/31) in those with a history of suspected signs/symptoms. Conclusion The epidemiological investigation indicates that the Andier Township of Minfeng County is an endemic area of visceral leishmaniasis.
    A Newly Identified Endemic Area of Visceral Leishmaniasis in Minfeng County of South Xinjiang Ⅱ. Investigation on Phlebotomine Vectors
    JINChang-fa*;ZUOXin-ping;GUDeng-an;Yisilayin·OSMAN;LANQin-xian;ZHANGYi;TONGShu-xiang;LIXiong;Ruziguli·ZHUMAHONG;Abulimiti;Muhetaer;Abuduwaili;Kuerban;NIUXin-ling
    2008, 26(2):  12-135. 
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    Objective To investigate the transmitting phlebotomine sandfly in Minfeng County,a newly-identified endemic area of visceral leishmaniasis in the south of Talim Pendi of Xinjiang. Methods Sandflies were collected using routine methods in and around the Yatonggusi village of Andier Township. The sandflies were identified to get their composition. Sandfly density was calculated following an observation at a given spot and time-period,and their appearance was recorded at night-time and day-time. Sandflies were dissected to analyze the gonotrophic cycle and to find infection of promastigotes. Results Phlebotomus wui was identified as the transmitting vector of visceral leishmaniasis in Andier Township,which occupied 99.2% of the sandflies collected. The first and second ten-day period of June was the first peak of its seasonal distribution. Analysis of the gonotrophic cycle revealed that Ph. wui was an exophilic species and appeared nocturnally for feeding with preference to human blood. Natural infection with promastigotes was found in 2 sandflies. Conclusion Phlebotomus wui is the transmitting vector for visceral leishmaniasis in the newly found endemic area of south Xinjiang.
    综述
    Human Serum Trypanosome Lytic Factor and Serum Resistance-Associated Protein of Trypanosome
    LIZhi;LUNZhao-rong*
    2008, 26(2):  13-140,. 
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    Human African trypanosomiasis is caused by the infection of Trypanosoma brucei gambiense or T.b. rhodesiense, while another morphologically identical subspecies, T.b.brucei, and other closely related species, T.equiperdum and T.evansi, are considered not infectious to human. This is highly related to the trypanosome lytic factor (TLF) found in normal human serum (NHS) and the serum resistance-associated (SRA) protein of trypanosomes infectious to human. We reviewed the research progress in TLF and its role in trypanosome lysis as well as the mechanism of SRA against the TLF.
    Research Progress on Chloroquine Resistance in Plasmodium falciparum and Resistance Reversal Agent
    QUANHong;TANGLin-hua*
    2008, 26(2):  14-145. 
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    Emergence and broad spread of chloroquine resistance urge human beings to change drug policy in malaria control and to find more effective new drugs. Nevertheless, chloroquine is still used in the treatment of falciparum malaria in some poor endemic regions due to economic and development reasons. It should be of great significance to understand the mechanism of chloroquine resistance and find the way to reverse it in order to bring chloroquine with high efficacy and low cost back to the first line of the combat to malaria. Advent and development of resistance reversal agents provide a new clue for this purpose. When used together with chloroquine, it can partly restore the efficacy of chloroquine in resistant Plasmodium falciparum. The article summarizes the research progress on chloroquine resistance in P. falciparum and resistance reversers.
    Diagnosis and Treatment of Major Protozoal Infections among Acquired Immune Deficiency Syndrome Patients
    SHENYin-zhong;LUHong-zhou*
    2008, 26(2):  15-148. 
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    Protozoal infection is one of the most important opportunistic infections among patients with acquired immune deficiency syndrome(AIDS). In order to enhance the knowledge of protozoal infections in AIDS, the current status of diagnosis and treatment of toxoplasmic encephalitis, cryptosporidiosis, microsporidiosis and isosporiasis was reviewed in this paper.
    研究简报
    Analysis of Karyotype and Preparation of C-band and G-band in Schistosoma japonicum
    HUANGLi-hong;ZHUGEHong-xiang*;ZHOUXia;LUXiang-bin;CHENMing-zhong;ZHANGHui-qin
    2008, 26(2):  16-151. 
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    Chromosomes of Schistosoma japonicum were prepared by usual air drying method, C-band and G-band were made by modified BSG method and enzyme digestion method respectively. Results showed that the karyotype of S. japonicun was 4m+6Sm+4St+2sex chromosome and the C-band formula was 2n= 5CI++4CI++3CI+2CT+2CT+.
    Polymorphism in the Partial Mitochondrial nad4 Gene of Schistosoma japonicum
    LIUWei;LIUYi;LINRui-qing;DAIRong-si;CHENGTian-yin;ZHUXing-quan*
    2008, 26(2):  17-153. 
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    The partial NADH dehydrogenase subunit 4 (pnad4) fragment was amplified by polymerase chain reaction (PCR) from Schistosoma japonicum collected from 4 areas in Hunan province, and the amplicons were cloned and sequenced. A 480 bp sequence was obtained and the nucleotide difference between samples from the four locations were 0.4%-1.3%. Low level of sequence variability between and within different populations of S.japonicum was detected. The results are worthwhile for further study on the population genetic structures of S.japonicum.
    Surgical Treatment of Patients with Cerebral Schistosomiasis
    WUMing-can*;CHENShi-jie;LUOGuo-cai;XUXian-ping;ZHANGZhi-wen;MINJie;YAOYuan
    2008, 26(2):  18-155. 
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    Clinical data from 48 cases with cerebral schistosomiasis, who received surgery, were analyzed retrospectively. Surgical treatment was performed when the patient had the following conditions: mass focus confirmed by CT scanning, cranial hypertension, or ineffective drug therapy, or indistinguishable from glioma by iconographic diagnosis. Treatment of praziquantel was given when the patients got improved. Among the 48 patients with surgical treatment, 35 cases recovered, 8 showed an exacerbation of hemiparalysis, 5 had hemianesthesia, and epilepsy occurred in 2 cases, no cases died during or after the operation. Surgery combined with praziquantel therapy has been an effective way for the treatment of cerebral schistosomiasis.
    In vitro Effect of Allitridium on the Ultrastructure of Acanthamoeba castellanii
    WANGYue-hua;ZHENGShan-zi*;LIShun-yu;CUIChun-quan
    2008, 26(2):  19-157. 
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    Acanthamoeba castellanii(T4)was cultured with different concentrations of allitridium for 24 hours, and examined by transmission electron microscope. The results showed that the ultrastructure of Acanthamoeba trophozoites was destroyed gently at concentration of 50 μg/ml allitridium and seriously destroyed under the concentration of 500 μg/ml, indicating that allitridium is effective in destroying Acanthamoeba.
    FTA/PCR Detection for Cryptosporidium Added in Beverage
    LIXiao-hong*;YANDong-li
    2008, 26(2):  20-159. 
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    An established FTA/PCR protocol was applied to detect Cryptosporidium oocysts which were added in beverage, and proved to be as sensitive as 1.0 oocyst/ml detected within 6 hours. The study indicates that this technique is sensitive, time-saving and easy to perform.
    Improved Preparation of Pure Alive Eggs of Schistosoma japonicum
    WANGYan-hai;PENGHong-juan*;GUJin-bao
    2008, 26(2):  21-封三. 
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    To prepare a large amount of pure alive Schistosoma japonicum eggs, rabbit was infected with 2000 cercariae and its liver was taken aseptically 38-45 days after infection and homogenized. The homogenate was screened through different sieves(60, 120, 200, 300, 360 meshes per inch respectively), and washed with 1.2% NaCl. The eggs and leftover were then digested with 0.25% trypsin for 2 hours, sieved over 360 meshes per inch and washed with RPMI 1640 medium. The collected eggs reached to (95.1±6.4)% of live eggs, with a high efficiency.