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Cloning and expression in Escherichia coli of a new gene of schistosoma japonicum encoding casein kinase Ⅱ beta subunit

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Chinese Medical Journal 2004年第9期117卷 1321-1325页

作者：彭寨玉余新炳吴忠道徐劲吴德李孜 Center for Disease Control and Prevention of Dongguan City Dongguan 523006 Guangdong China

Background Nowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase Ⅱ beta subunit, of schistosoma japonicum (S. japonicum) and express it in Escherichia coli (***).Methods The ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3’RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into *** JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot. Results A 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase Ⅱ beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in *** *** recombinant protein could be recognized by the S. japonicum infected rabbit serum. Conclusion The full-length cDNA sequences encoding S. japonicum casein kinase Ⅱ beta subunit were firstly sequenced, cloned, and expressed in ***.

关键词： schistosoma japonicum casein kinase Ⅱ beta subunit cloning expression

DNA vaccine pCD-Sj32 and its efficacy of protective immunity against infection of schistosoma japonicum

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Chinese Medical Journal 1999年第8期112卷 5-8页

作者：石佑恩谢来平李传明韩家俊 Department of Parasitology Tongji Medical University Wuhan 430030 China Department of Immunology University of Alabama Briminghan al(USA)

Objective To study protective immunity afforded by murine immunization with DNA vaccine of schistosoma japonicum (S. japonicum) as measured by reduction in worm burden and host antibody, *** DNA vaccine pCD Sj32 was constructed. identificated and expressed. pCD Sj32 could induce substantial protective immunity against infection of S. japonicum in BALB/c mice. The best efficacy can be produced with one injection of 100?μg DNA into the quadriceps muscle, combined with challenge for 8 weeks after immunization. T lymphocyte subsets of CD 8 +, IL 2. TNF and IFN γ of experimental animal could play important roles in regulating immune functions of schistosomiasis. Results High titre of specific antibody IgG could be induced by vaccinated with pCD Sj32, and antibody can mediate macrophage to produce ADCC effects in vitro. Conclusion pCD Sj32 may represent a new approach to developing subunit vaccine.

关键词： schistosoma japonicum DNA vaccine protective immunity

Partial protection induced by phage library-selected peptides mimicking epitopes of schistosoma japonicum

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Chinese Medical Journal 2003年第1期116卷 138-141页

作者：欧阳理易新元Department of Pathogen Biology 曾宪芳Department of Pathogen Biology 周金春Department of Pathogen Biology 王庆林Department of Pathogen Biology Larry Mc Reynolds Department of Pathogen Biology New England Biolabs

Objective To obtain peptide mimicking epitopes of schistosoma japonicum (***) through screening of a phage peptide library and to test their potential for induction of protection. Methods *** infected sera from Microtus fortis (IMFS) and normal sera from Microtus fortis (NMFS) were used respectively to screen a 12-mers random peptide library by testing the reactivity of *** serum with the phagotopes. After three rounds of biopanning, the pooled phages were used to immunize mice, after which challenge infection was performed. Results Of 12 randomly picked clones, 10 clones selected using IMFS and 7 clones selected using NMFS were shown to be antigenic. Significant reduction in adult worms (22.6%) and a high reduction (68.9%) in liver eggs were achieved following immunization with phages screened with IMFS. However, no protection was elicited by those selected with NMFS. Conclusion The results show that the phagotopes are both antigenic and immunogenic, suggesting a potential use of phage displayed peptide as novel vaccines against S. japonicum.

关键词： peptide library epitopes schistosoma japonicum

Recombinant protein schistosoma japonicum-derived molecule attenuates dextran sulfate sodium-induced colitis by inhibiting miRNA-217-5p to alleviate apoptosis

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World Journal of Gastroenterology 2021年第46期27卷 7982-7994页

作者： Li-Chao Zhang Xiao-Ying Wu Rui-Bing Yang Fang Chen Jia-Hua Liu Yun-Yi Hu Zhong-Dao Wu Li-Fu Wang Xi Sun Department of Parasitology of Zhongshan School of Medicine Sun Yat-sen UniversityGuangzhou 510000Guangdong ProvinceChina Department of Gastroenterology Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhou 510630Guangdong ProvinceChina School of Medicine South China University of TechnologySouth China University of TechnologyGuangzhou 510000Guangdong ProvinceChina Key Laboratory of Tropical Disease Control Ministry of EducationSun Yat-sen UniversityGuangzhou 510080Guangdong ProvinceChina

BACKGROUND Inflammatory bowel disease(IBD)affects millions of people worldwide and has emerged as a growing problem in industrialized *** lack of therapeutic targets has limited the treatment of *** found that parasitic nematode infections can ameliorate clinical and experimental *** previous study found that rSj16,a 16-kDa secreted protein of schistosoma japonicum produced by Escherichia coli,has protective effects on dextran sulfate sodium(DSS)-induced colitis in *** is an important factor in the pathogenesis of ***,it is not clear whether the effect of rSj16 on colitis is related to *** To investigate whether the protective effects of rSj16 on colitis is related to apoptosis and its *** In-vivo,colitis was induced by *** severity of colitis was *** was used to detect the changes of apoptosis-related genes in colon tissues.Q-PCR was used to detect the changes of miRNA-217-5p and ***-vitro,WB was used to detect the changes of apoptosis-related genes in intestinal epithelial *** staining and flow cytometry were used to detect cell *** rSj16 attenuates clinical activity in DSS-induced colitis *** staining and WB results showed that apoptosis was increased in colon tissue after treatment with DSS,and the apoptosis of colon tissue was significantly reduced after treatment with *** with normal mice,the expression of miR-217-5p was increased in colon tissue of DSS-induced colitis *** addition,the miR-217-5p target gene hnf1b was decreased after administration of *** treatment with rSj16,the expression of miR-217-5p was decreased and the expression of HNF1B was increased compared with the DSS-treated *** Etoposide was used in combination with miR-217-5p mimic on MODE-K cells,the expression of cleaved-Caspase-3 and Bax was increased,and Bcl-2 was decreased compared with only Etoposide treatment,the expression of HNF1B was signif

关键词： schistosoma japonicum rSj16 Inflammatory bowel disease Apoptosis miRNA-217-5p

Real-time PCR diagnosis of schistosoma japonicum in low transmission areas of China

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Infectious Diseases of Poverty 2018年第1期7卷 87-97页

作者： Pei He Catherine A.Gordon Gail M.Williams Yuesheng Li Yuanyuan Wang Junjian Hu Darren J.Gray Allen G.Ross Donald Harn Donald P.McManus Hunan Institute of Parasitic Diseases YueyangPeople’s Republic of China Molecular Parasitology Laboratory Infectious Diseases DivisionQIMR Berghofer Medical Research InstituteBrisbaneAustralia Discipline of Epidemiology and Biostatistics School of Population HealthUniversity of QueenslandBrisbaneAustralia Research School of Population Health The Australian National UniversityCanberraAustralia Menzies Health Institute Queensland Griffith UniversityGold CoastQLDAustralia University of Georgia College of Veterinary MedicineAthensGAUSA.

Background:Schistosomiasis in the People’s Republic of China(PRC)can be traced back to *** the past 60 years,the Chinese government has made great efforts to control this persistent disease with elimination slated by 2020 through the implementation of a comprehensive control *** strategy aims to reduce the role of bovines and humans as sources of infection as a pre-requisite for elimination through transmission *** goal of elimination will be achievable only by the implementation of a sustainable surveillance and control system,with sensitive diagnosis a key feature so that the true disease burden is not *** used diagnostics lack the necessary sensitivity to accurately determine the prevalence of schistosoma japonicum infection in areas with low infection *** is of critical importance to find and treat people and to identify animals with low-level infections if the National Control Programme for China is to achieve schistosomiasis ***:We evaluated a real-time polymerase chain reaction(qPCR)assay using 633 human stool samples collected from five villages in Hunan,Anhui,Hubei,and Jiangxi provinces,and 182 bovine(70 cattle and 112 buffalo)stool samples obtained from four villages in Hunan,Anhui,and Jiangxi provinces in the *** stool samples were subjected to the miracidium hatching test(MHT,a diagnostic procedure used in the National Schistosomiasis Control Programme)and the qPCR *** positive by MHT were subjected to either the Kato-Katz technique for humans,or the formalin-ethyl acetate sedimentation-digestion(FEA-SD)procedure for bovines,to determine infection ***:The qPCR assay exhibited a high level of sensitivity in the detection of *** *** both the human and bovine samples,a significantly higher prevalence was determined using the qPCR assay(11.06%humans,24.73%bovines)than with the MHT(0.93%humans,7.69%bovines).The animal contamination

关键词： Schistosomiasis schistosoma japonicum China Real-time PCR Miracidium hatching test Kato-Katz FEA-SD Bovine Human

Assessment of morbidity due to schistosoma japonicum infection in China

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Infectious Diseases of Poverty 2014年第1期3卷 40-55页

作者： Ming-gang Chen National Institute of Parasitic Diseases Chinese Center for Disease Control and PreventionShanghaiPeople’s Republic of China WHO Collaborative Center for Malaria Schistosomiasis and FilariasisShanghaiPeople’s Republic of China

This paper presents a historical assessment of morbidity due to the schistosoma japonicum infection in *** to the socio-economic situation,which did not allow for a control program to be implemented until the early 1950s,morbidity was serious and mortality was high before *** on a few investigations and published papers,it can be said that the disease caused millions of deaths,and destroyed numerous families and *** the 1950s,there has been a national control program,intensive control and prevention work has been carried out,and consequently the disease is being *** present,both the prevalence and the morbidity of the disease have been decreasing *** morbidity of the three phases of the disease is outlined in this *** higher morbidity is seen in the acute and advanced phases of the *** four major forms of advanced schistosomiasis i.e.,ascites,megalosplenia,dwarfism,and colonic tumoroid proliferation,are outlined with their characteristic clinical presentations;their proportions are different during various periods of the national control *** schistosomiasis and the relationship between the *** infection and colorectal cancer are also ***-transmission schistosomiasis is briefly discussed(which can happen even if the disease reaches the criteria of elimination,and the infection and transmission have stopped,but yet it still develops).The problem of mammalian reservoir hosts of *** makes the epidemiology and control of schistosomiasis in China even more complicated and arduous,and the control progress in animal reservoirs is briefly presented.

关键词： schistosoma japonicum Schistosomiasis Morbidity China

Pharmacological and immunological effects of praziquantel against schistosoma japonicum:a scoping review of experimental studies

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Infectious Diseases of Poverty 2018年第1期7卷 98-112页

作者： Shu-Hua Xiao Jun Sun Ming-Gang Chen National Institute of Parasitic Diseases Chinese Center for Disease Control and PreventionKey Laboratory of Parasite and Vector BiologyMinistry of HealthWHO Collaborating Centre for Tropical DiseasesShanghai 200025People’s Republic of China Institute for Infectious Disease and Vaccine Development Tongji University School of MedicineShanghai 200092People’s Republic of China

Background:Chemotherapy for schistosomiasis has been around for 100 *** the past century,great efforts have been made to develop new antischistosomal drugs from antimonials to nonantimonials,and some of these have been used extensively in clinical *** the exception of a few drugs,such as oxamniquine and metrifonate,most of the antischistosomals developed in the pre-praziquantel period have variable limitations with respect to safety and *** oxamniquine and metrifonate have been used for schistosomiasis control,they are only effective against schistosoma mansoni and ***,***,praziquantel is the only drug used for treatment of all five species of human *** this review,the pharmacological and immunological effects of praziquantel against *** are summarized and *** text:From the end of the 1970s until the 2000s,scientists have conducted a series of experimental studies on the effects of praziquantel against *** have included examining its unique pharmacological action on schistosomes,the characteristics in susceptibility of the different developmental stages of schistosomes to the drug,the relationship between plasma concentration of the drug and efficacy,the impact of host factors on cidal action of the drug,prevention and early treatment of schistosomal infection,as well as praziquantel-resistant ***:The effects of praziquantel against ***,as elucidated by the experimental studies that are reviewed in this paper,may have some reference significance for the development of new antischistosomals.

关键词： Praziquantel Pharmacological action Immunological action schistosoma japonicum Experimental study Review

Effect of RNA interference on WD101 gene of schistosoma japonicum

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Asian Pacific Journal of Tropical Medicine 2018年第2期11卷 136-140页

作者： Peng Zhang Gang Feng Wei-Na Zhang Ying-Ying Zhang Chun-Sheng Liu Department of Clinical Laboratory Yijishan Hospital of Wanan Medical College

Objective: To study the effect of RNA interference(RNAi) on WD101 gene and its effect on the expression of WD101 mRNA and protein in schistosoma ***: Doublestranded RNA(dsRNA) WD101 gene and control gene(lacZ) were generated by in vitrotranscription and transfected into mechanically transformed *** total RNA and protein were isolated simultaneously using TRIzol *** expression levels of mRNA and the protein were determined by quantitative real-time PCR(qPCR) and Western blotting, *** injected dsRNA-electroporated schistosomula into BALB/c mouse six weeks, the male and female reproductive organs were observed and measured under the confocal laser scanning ***: After 1, 3 and 5 d of RNAi, WD101 mRNA level was decreased by 15%, 39%, and 58% in experiment group compared to that in control group; meanwhile, WD101 protein level was decreased by 11%, 28%, and 43% in experiment group compared to that in control *** were significantly more sperms in testicular lobes in experiment group than that in control group, while there were no significant differences in terms of ovary and vitelline glands between two ***: The ds WD101-RNAi can effectively induce suppression of WD101 gene expression at both mRNA and protein ***101 gene might be a reproduction-related gene in schistosoma japonicum.

关键词： schistosoma japonicum RNA interference WD101

Systems biomedical analysis of schistosoma japonicum

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Frontiers of Medicine 2010年第2期4卷 157-165页

作者： Ze-Guang HAN Shanghai-MOST Key Laboratory for Disease and Health Genomics Chinese National Human Genome Center at ShanghaiShanghai 201203China Rui-Jin Hospital Shanghai Jiao Tong University School of MedicineShanghai 200025China

Human schistosomiasis,caused mainly by three principal species including schistosoma japonicum,***,and ***,remains a major public health concern *** is prevalent in southern China,being a major disease risk for 66 million *** bloodﬂuke has a complex life cycle for survival:as a free-living form in fresh water and as a parasite in the snail intermediate and vertebrate deﬁnitive ***-based biomedical analyses,including genomic,transcriptomic,proteomic and metabonomic approaches,have been performed on the *** comprehensive investigations have not only char-acterized the genomic features but also chartered gene and protein expression proﬁles across genders and develop-mental *** integration of the huge information will lay a global and solid foundation for the molecular architecture of the biology,pathogenesis,and host-parasite interactions of the human bloodﬂuke,which will facilitate the development of a new antischistosomal vaccine and drugs as well as diagnostic markers for the treatment and control of schistosomiasis.

关键词： schistosoma japonicum systems biomedical analysis