Yong-hua Zhou, Ying-ying Yang, Xiao-lin Fan, Xue Sai
{"title":"[The role of Th17/Treg imbalance in liver fibrosis in schistosomiasis].","authors":"Yong-hua Zhou, Ying-ying Yang, Xiao-lin Fan, Xue Sai","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Liver fibrosis in schistosomiasis is a serious pathological consequence from immune reactions to schistosome infection. The progression of liver fibrosis depends on the state of immune response. Recent studies have found that Th17 and Treg cells are two subsets of CD4+T cells. The Th17 cells are mainly involved in inflammatory responses, while the Treg cells mainly mediate downregulation of the responses. Under normal conditions, the differentiations of the two subsets are inhibited by each other, and they function oppositely. The balance between Th17 and Treg cells, as well as the balance between them, play an important role in the maintenance of homeostasis and are involved in inflammatory responses, tissue trauma, fibrosis and development of many diseases. This paper reviews the role of Th17/Treg cells and their imbalance in liver fibrosis in schistosomiasis.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"89-92"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36417921","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiao-dong Zhan, Bin-bin Duan, Ning Tao, Chao-pin Li
{"title":"[The effect of Dermatophagoides pteronyssinus group 2 allergen T cell fusion epitope on STAT6 signaling in mice with asthma].","authors":"Xiao-dong Zhan, Bin-bin Duan, Ning Tao, Chao-pin Li","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the alterations of signal transducer and activator of transcription factor 6(STAT6) signaling in a mouse model of asthma receiving treatment with Dermatophagoides pteronyssinus group 2 allergen(Der p 2) T cell fusion epitope and the mechanisms of the specific immunotherapy.</p><p><strong>Methods: </strong>Thirty mice were randomly divided into three groups by the random number table method: the asthma group, the treatment group receiving immunotherapy with Der p 2 T cell fusion epitope, and the negative control group (PBS group)(n = 10 in each group). Mice in the asthma and the treatment groups received intraperitoneal (i. p.) injection of 100 μl Der p 2 solution [PBS containing 100 μg/ml Der p 2 and 2% Al(OH)3] on days 0,7 and 14, respetively, while mice in the PBS group received same volume of PBS containing 2% Al(OH)3. From day 21, 30-min steam inhalation of 0.5 μg/ml Der p 2 was applied to the asthma and treatment groups (once daily for 7 successive days), and the PBS group inhaled same volume of PBS. From day 25 to day 27, the mice in the treatment group received i. p. injection of 200 μl of Der p 2 T cell fusion epitope (100 μg/ml) while the PBS and the asthma groups received the same volume of PBS. Mice were sacrificed at 24 h after the last inhalation, the bronchoalveolar lavage fluid (BALF) collected, and the total protein was extracted from the lung tissue. The levels of IFN-γ, IL-4 and IL-13 in BALF were determined by ELISA. The expression of STAT6 and phosphorylated STAT6 (p-STAT6) in the lung tissue was detected by Western blotting. Data were analyzed with the one-way variance analysis (ANOVA) method.</p><p><strong>Results: </strong>The level of IFN-γ in the treatment group[(234.40 ± 24.46) pg/ml] was significantly higher than that in the asthma group[(155.80 ± 20.53) pg/ml](P < 0.01). The levels of IL-4 and IL-13 in the treatment group [(30.00 ± 5.50) pg/ml and (174.50 ± 25.99) pg/ml, respectively] were both significantly lower than those in the asthma group[(53.28 ± 8.26) pg/ml and (308.10 ± 28.32) pg/ml, respectively](P < 0.01). Similarly, the levels of STAT6 and p-STAT6 in the treatment group(0.803 ± 0.221 and 0.966 ± 0.323, respectively) were both significantly lower than those in the asthma group (1.669 ± 0.296 and 1.735 ± 0.298, respectively)(P < 0.01).</p><p><strong>Conclusion: </strong>The Der p 2 T cell fusion epitope may function through inhibiting STAT6 to treat asthma in mice.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"19-23"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36415125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guang Chen, Lei Liu, Fang-fang Wang, Sheng Bi, Lan Luo, Ju-xiang Su, Hui-ming Zhang, Lian-shun Cai, Zi-lin Gong
{"title":"[The regulatory effect of dendritic cells on Th17 cell differentiation and function in mouse infected with Plasmodium yoelii].","authors":"Guang Chen, Lei Liu, Fang-fang Wang, Sheng Bi, Lan Luo, Ju-xiang Su, Hui-ming Zhang, Lian-shun Cai, Zi-lin Gong","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To investigate the regulatory effect of dendritic cells (DCs) on Th17 cell differentiation and function during mouse infection with Plasmodium yoelii 17XL strain (Py17XL) and the underlying mechanisms.</p><p><strong>Methods: </strong>Twelve female BALB/c mice were randomly assigned into the infection group (Py17XL), the TLR4 blocking group (Py17XL + TLR4), TLR9 blocking group (Py17XL + TLR9), and TLR4 and TLR9 combined blocking group (Py17XL + TLR4+TLR9)(n=3 in each group). Mice in the Py17XL + TLR4 or the Py17XL + TLR9 group received intraperitoneal injection of 10 μg anti-TLR4 or 50 μg anti-TLR9 antibody (both 0.4 ml) to block DCs function at one day before infection. The Py17XL group received same volume of PBS. All groups were then given intraperitoneal injection of 1×10(6) red blood cells (RBCs) infected with Py17XL. The RBC infection rate was calculated on days 0, 3 and 5 after infection, and spleen cell suspension was prepared, in which the CD11c+TLR9+ and Th17 cells were counted by flow cytometry. The levels of IFN-γ and IL-10 in supernatant of spleen cell culture were determined by ELISA.</p><p><strong>Results: </strong>Flow cytometry showed that DCs were successfully blocked. On day 5 after infection, 28%,29%, 31% and 16.3% mice showed parasitemia in the Py17XL group, the Py17XL + TLR4 group, the Py17XL + TLR9 group, and the Py17XL + TLR4 + TLR9 group, respectively, and on day 7, the proportions were 43.3%, 47.5%, 32.5% and 8%. Mice in the Py17XL group and the Py17XL + TLR4 group all died, while those in other groups began to die from day 6. There was a slow rise of parasitemia rate in the Py17XL + TLR9 group and the Py17XL + TLR4 + TLR9 group compared with the Py17XL group, with a significant extension of survival to days 11 and 15. Results of flow cytometry showed that the proportions of Th17 cells were 1.2% and 1.44% in the Py17XL + TLR9 group and the Py17XL + TLR4 + TLR9 group on day 5, both sighificantly decreased compared with the Py17XL group (1.9%)(P < 0.05, P < 0.01). ELISA revealed that the levels of IFN-γ and IL-10 on day 5 in the Py17XL + TLR4 + TLR9 group [(232.4 ± 15.5) pg/ml and(1791.2 ± 58.2) pg/ml, respectively] were significantly higher than those in the Py17XL group[(90.7 ± 50.1) pg/ml and (962.6 ± 409.0) pg/ml](P < 0.05, P < 0.01).</p><p><strong>Conclusions: </strong>The differentiation and function of Th17 cells are regulated by DCs during Py17XL infection. Blockade of DCs decreases parasitemia and extends lifetime of mice. Further studies are needed to clarify the exact mechanisms.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"8-12"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36417709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fang-zhen Xiao, Xiu-qing Peng, Guo-ying Xu, Yang Chen, Dai-hua Lin, Yan-qin Deng
{"title":"[Investigation and genetic identification on Babesia infection in rodents in some areas of Fujian Province].","authors":"Fang-zhen Xiao, Xiu-qing Peng, Guo-ying Xu, Yang Chen, Dai-hua Lin, Yan-qin Deng","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To explore the status of Babesia infection in rodents and the genetic characteristics of Babesia spp. in Fujian Province.</p><p><strong>Methods: </strong>Rodents were captured by the night trapping method in Shaowu, Qingliu, Shunchang, Yong’an, Changle and Youxi during 2014-2015. The rodent species was identified, and information on the time and place of capture, species and sex of rodents was recorded. Blood samples was collected, in which the fragment of 18S rRNA gene of Babesia spp. was amplified by PCR. The PCR products were sequenced and the phylogenetic tree was constructed for homology analysis. Data on positive rate were analyzed with Chi-square or Fisher exact test.</p><p><strong>Results: </strong>Two hundred and nine rats were captured, comprising of 71 domestic and 138 wild rats. The overall positive rate was 9.6%(20/209). The positive rate in domestic rats was 2.8%(2/71), including one Rattus norvebicus and one Rattus flavipectus. The positive rate in wild rats was 13.0%(18/138), including 13 Bandicota indica, one Rattus losea, 2 Rattus confucianus and 2 Rattus fulvescens. The positive rate was significantly higher in wild rats than in domestic rats (P < 0.05). The Youxi region had the highest positive rate(14.9%,13/87), followed by Yong’an(13.6%, 3/22), and no positive rat was found in Qingliu. The positive rate in the male rats was 7.9%(9/114), and that in the females was 11.6%(11/95). The positive rate was highest in adult rats (10.4%,18/173), followed by young ones (6.3%,2/32). No positive rat was found in old rats. There was no significant difference in positive rate among different regions, between male and female rats, or among different ages (P > 0.05). The sequences of PCR products had a 100% homology. The BLAST results revealed the species to be Babesia microti. The phylogenetic tree showed that the sample sequence was the most homologous with Babesia microti from Zhejiang Province(GenBank Accession No: JQ609305).</p><p><strong>Conclusion: </strong>There occurs Babesia microti infection in rats in part areas of Fujian Province. The positive rate was higher in wild rats than in domestic rats.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"63-7"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36419554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Li Li, Yang Liu, Guo-jun Xu, Tao Yu, Yan Zou, Xiao-hong Wu, Bo Zhong
{"title":"[Epidemiological characteristics of imported malaria in Sichuan Province in 2015].","authors":"Li Li, Yang Liu, Guo-jun Xu, Tao Yu, Yan Zou, Xiao-hong Wu, Bo Zhong","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the epidemiological characteristics of imported malaria in Sichuan Province in 2015 in the aim of providing scientific basis for malaria control.</p><p><strong>Methods: </strong>The epidemiological data of malaria in Sichuan Province reported through the Infectious Disease Reporting and Management System of China Information System for Disease Control and Prevention in 2015 were collected and analyzed.</p><p><strong>Results: </strong>In 2015,290 malaria cases were reported in Sichuan Provinc, consisting of 158 falciparum malaria cases (54.5%),107 vivax malaria cases (36.9%),14 ovale malaria cases (4.8%), one quartan malaria case(0.3%), and 10 mixed infections of vivax malaria and falciparum malaria (3.5%). Five cases of falciparum malaria died. The reported cases were all imported, with a major source of Africa (271, 93.4%), in which Ethiopia (83) and Angola (49) were two major sources. The cases were reported continuously from January to December, with the majority(139, 47.9%) being reported in December, August, June and July. The cases distributed mainly in Guangan, Chengdu, Nanchong, Mianyang, Deyang, Luzhou and Suining(243, 83.8%). Among the 209 cases who first visited medical units after onset, 118 cases were diagnosed as malaria, the misdiagnosis rate at first visit was 43.5%(91/209).</p><p><strong>Conclusion: </strong>The malaria cases reported in Sichuan Province in 2015 are all imported from overseas, mainly infected with P. falciparum and P. vivax, and imported mostly from Africa. There is a high rate of misdiagnosis in medical units in Sichuan Province.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"75-9"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36417918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lei Lei, Zhi-gui Xia, Zhi-hong Li, Yan-feng Gong, Ning Xiao
{"title":"[A retrospective analysis on the diagnosis and reporting of imported malaria in Jiangxi Province during 2012-2015].","authors":"Lei Lei, Zhi-gui Xia, Zhi-hong Li, Yan-feng Gong, Ning Xiao","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To provide scientific basis for malaria surveillance in the elimination phase by retrospectively analyzing the diagnosis and reporting of imported malaria in Jiangxi Province.</p><p><strong>Methods: </strong>Data on malaria endemic situation and individual cases during 2012-2015 were collected through the National Information Management System for Infectious Diseases and the Report and Information Management System for Parasitic Diseases Control and Prevention. Detailed information on primary medical units, laboratory testing units, reporting units, diagnostic methods, time from onset to first medical visit, time from first medical visit to reporting, and time from onset to reporting was analyzed with the descriptive analysis method.</p><p><strong>Results: </strong>A total of 207 malaria cases were reported during 2012-2015 in Jiangxi, all were imported cases and 96.62%(200/207) were diagnosed with laboratory tests. The main primary medical units were found to be county-level (29.95%, 62/207) and prefecture-level (25.60%, 53/207) medical institutions, while the main laboratory testing units were prefecture-level medical institutions(35.27%,73/207) and county-level CDCs (20.29%, 42/207). There was a significant difference in the proportion of different laboratory testing units among the years(P < 0.05). The median time from onset to first medical visit was 1 d (0-149 d), from first medical visit to reporting was 3 d (0-144 d), and from onset to reporting was 5 d (0-149 d).</p><p><strong>Conclusions: </strong>The first visit and the laboratory testing of malaria cases mainly occur in the prefecture-level and county-level medical institutions.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"80-4"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36417919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"[Proteomics analysis of Paragonimus skrjabini].","authors":"An-mei Li, Ma-li Wu, Yu-ting Huang, Zhi-lai Guo","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To perform a proteomics analysis for metacercariae, juvenile and adult worms of Paragonimus skrjabini.</p><p><strong>Methods: </strong>Crabs were collected in P. skrjabini endemic areas of Kaiyang and Bijie in Guizhou Province. Metacercariae were isolated, placed in PBS, and were used to infect three SD rats(10 metacercariae/rat) by intragastric administration and infect three male dogs(20 metacercariae/dog) through feeding. The rats were sacrificed at 1 month after infection to obtain juvenile worms. The dogs were sacrificed at 3 months after infection to obtain adult worms. The metacercariae, juvenile and adult worms were lysed, and total protein was extracted by ultrasonication. The total protein content was determined by Bradford method and separated by SDS-PAGE and two dimensional gel electrophoresis. Images of two dimensional gel electrophoresis were analyzed using the PDquest 8.0 software. The dots with difference were digested and analyzed with mass spectrometry. Finally, online searches in NCBI and local databases were performed.</p><p><strong>Results: </strong>Results of SDS-PAGE showed that the total protein of metacercariae, juvenile and adult worms was concentrated within Mr of 25 000-116 000. Fifty-one protein dots with difference were found by two dimensional gel electrophoresis, comprising of 20 dots for metacercariae, 25 for juvenile worms and 6 for adult worms. Thirty-six peptide sequences of metacercariae and juvenile worms were analyzed. They were basically determined to be Achrornobacter lyticus protease Ⅰ(a lysine-specific serine protease), ascorbate reductase protein, glutathione s-transferase DHAR2 analogue, heat shock proteins Hsp82 and Hsp96-β, actin, cystatin, etc., by online searches, and cysteine, actin and heat shock protein by local searches in the diginea database(downloaded to a local computer from NCBI). Mass spectrometry was not performed for adult worms, as the variation of grayscale value between their spots was far less than those for metacercariae and juvenile worms.</p><p><strong>Conclusion: </strong>One difference is that the metacercariae of P. skrjabini have actin, while the juvenile worms have detoxification proteins and stress proteins. But they both have hydrolases and cysteine enzymes.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"43-7"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36419568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"[Component analysis of excretory/secretory protein from Trichinella spiralis adult worm].","authors":"Xiao-di Yang, Zhi-yong Tao, Yang Cheng, Qi Wu, Xiao-li Wang, Di Song, Lan-song Xu, Ren-min Xue, Xue-lian Chang, Hui Zhang, Rui Wang, Xing-zhi Chen, Qiang Fang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the component of adult worm excretory/secretory protein(AWESP) from Trichinella spiralis using the shotgun method, and find out the active component underlying its regulatory effect on colitis in humans.</p><p><strong>Methods: </strong>The T. spiralis AWESP was prepared, separated by SDS-PAGE, lysed with trypsin, and analyzed by shotgun LC-MS/MS. The protein components were determined with the Masco software and classified using the Gene Ontology(GO) method in cellular components, molecular functions, and biological processes.</p><p><strong>Results: </strong>The AWESPs isolated by SDS-PAGE had a Mr of 15 000-116 000. A total of 280 proteins were revealed by LC-MS/MS, of which 96 were identified by Masco software, 98 were putative, and the remaining 86 were unclear. Preliminary results showed that 4 proteins had regulatory potential for colitis, including cysteine protease inhibitor, serine protease, 53 000 excretory/secretory antigen, and glutathione-S-transferase. GO enrichment analysis showed that the identified proteins had 104 different molecular functions, involved in 363 biological processes.</p><p><strong>Conclusion: </strong>As revealed by the Masco software, T. spiralis AWESP has complex components and 96 have been identified in this study. Four of them are preliminarily shown to be associated with the anti-colitis effect of T. spiralis.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"24-9"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36415126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"[Cloning of galectin-1 gene of Angiostrongylus cantonensis and testing the agglutination property of the galectin-1 protein].","authors":"Xing-pan Li, Meng-jing Zhao, Xiao-meng Shi, Lan-zhu Yan, Bao-long Yan, Hui-cong Huang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To clone and express the galectin-1 gene of Angiostrongylus cantonensis, and test the agglutination property of its protein.</p><p><strong>Methods: </strong>The three-dimensional structure of galectin-1 was analyzed with Swiss Model. Total RNA was extracted from male worms of A. cantonensis. Primers were designed for galectin-1 based on its coding region (GenBank Accession No. JN133961.1). RT-PCR was performed, and the PCR products were subcloned to pCold Ⅲ plasmid and transduced into Escherichia coli BL21 strain. The recombinant plasmid was extracted from positive clones on LB plate containing 100 μg/ml Kanamycin, and validated with double digestion, PCR identification and sequencing. The confirmed positive clones of E. coli BL21 with the recombinant plasmid were grown in LB medium containing ampicillin (100 μg/ml, 100 μl). IPTG was added to induce expression of the plasmid. The galectin-1 recombinant protein was purified with Ni-NTA beads, and analyzed with SDS-PAGE and Western blotting using anti-serum of mouse immunized with whole worms of A. cantonensis. The agglutination reaction with red blood cells in fresh blood of ICR mouse was observed for the 10-fold serial dilutions of recombinant proteins (5.55 × 10(-1)-5.55 × 10(-5) ng/μl).</p><p><strong>Results: </strong>The Swiss Model analysis showed that the functional galectin-1 had a non-dimeric form. As was expected, the RT-PCR products had a size of 850 bp. Results of double digestion, PCR and sequencing showed successful construction of the pCold Ⅲ-galectin-1 plasmid. SDS-PAGE revealed expression of soluble recombinant fusion protein with molecular weight of ~36 000. Western blotting showed that the galectin-1 protein was recognized by mouse anti-serum. In addition, the minimun concentration of galectin-1 that showed significant agglutination reactions with mouse red blood cells was 5.55 × 10(-4) ng/μl.</p><p><strong>Conclusion: </strong>The galectin-1 clone can be expressed in the pCold Ⅲ plasmid, and its protein product has agglutination property.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"48-52"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36419571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ying Dong, Ai-ming Sun, Meng-ni Chen, Yan-chun Xu, Xiang-hua Mao, Yan Deng
{"title":"[Polymorphism analysis of the block 5 region in merozoite surface protein-1 gene of imported and local Plasmodium vivax in Yunnan Province].","authors":"Ying Dong, Ai-ming Sun, Meng-ni Chen, Yan-chun Xu, Xiang-hua Mao, Yan Deng","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the sequence of Plasmodium vivax merozoite surface protein-1(PvMSP-1) and allele polymorphism in imported and local vivax malaria parasites in Yunnan Province.</p><p><strong>Methods: </strong>Blood samples on filter paper were collected from imported and local vivax malaria cases in Yunnan Province during August 2012 and September 2015 and information of epidemiological history was recorded. Plasmodium DNA was extracted by a DNA extraction kit, and the block 5 region in PvMSP-1 gene was amplified by PCR. The PCR products were sequenced and blasted with reference sequences M75674, AAN86237, M60807, ABJ53045, AAN86238 and BAA18977. The sequence polymorphism in block 5 region of PvMSP-1 was analyzed with MEGA 5.04 and Arlequin3.5.1 softwares. The conserved sites, genetic distances among sequences and Shannon-wiener index among alleles were calculated. The clustering tree was drawn according to the genetic distances between the amino-acid sequences.</p><p><strong>Results: </strong>A total of 847 blood samples were collected from the malaria cases, comprising of 61 samples from local cases, 66 from imported cases from Africa, and 720 from Myanmar. The block 5 region in PvMSP-1 was successfully amplified in 278 samples, and sequencing was successfully made in 206 of them. The peptide coded by the block 5 region had a length of 193 to 222 aa. The amino acid sequence alignment showed that in 206 samples the proportion of genotypes of Sal-1, Belem and Recombine was 59.2%(122/206),23.3%(48/206) and 17.5%(36/206), respectively. The proportion of Sal-1 genotype in imported cases from Myanmar and Africa and in local cases was 58.8%(104/177),73.3%(11/15) and 50%(7/14), respectively. The genotypes Sal-1, Belem and Recombine had 51, 9 and 6 different alleles. The 66 alleles had a Shannon Wiener index (H’) of 0.955 and an expected heterozygosis (He) of 0.567. The 206 DNA sequences had a 665-bp homologous locus, comprising of 75 conserved sites (11.3%,75/665) and 590 variable sites (88.7%, 590/665). The genetic distances between sequences were all less than 0.4. The clustering analysis showed that the 206 sequences were clustered into two categories with three branches. The homology of Recombine with Belem genotype was 91%-92%, higher than with Sal-1 genotype (82%-83%).</p><p><strong>Conclusion: </strong>The block 5 region in PvMSP-1 gene from local and imported Plasmodium vivax in Yunnan Province has varied forms of alleles, and the Sal-1 genotype is predominant among the three genotypes.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"35 1","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36417705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}