35 results on '"Gu, Xiaobin"'
Search Results
2. Comprehensive molecular characterization of complete mitogenome assemblies of 33 Eimeria isolates infecting domestic chickens
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Zhou, Xuan, Wang, Lidan, Zhu, Pengchen, Yang, Zijiang, Wang, Zhao, Chen, Yijun, Gu, Xiaobin, He, Ran, Xu, Jing, Jing, Bo, Yang, Guangyou, Chen, Shun, Wu, Shuangyang, and Xie, Yue
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- 2023
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3. A novel cysteine protease inhibitor in Baylisascaris schroederi migratory larvae regulates inflammasome activation through the TLR4–ROS–NLRP3 pathway
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Xu, Jingyun, Gu, Xiaobin, Xie, Yue, He, Ran, Xu, Jing, Xiong, Lang, Peng, Xuerong, and Yang, Guangyou
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- 2022
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4. Identification, characterization and functional analysis of gonadal long noncoding RNAs in a protogynous hermaphroditic teleost fish, the ricefield eel (Monopterus albus)
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He, Zhi, Ye, Lijuan, Yang, Deying, Ma, Zhijun, Deng, Faqiang, He, Zhide, Hu, Jiaxiang, Chen, Hongjun, Zheng, Li, Pu, Yong, Jiao, Yuanyuan, Chen, Qiqi, Gao, Kuo, Xiong, Jinxin, Lai, Bolin, Gu, Xiaobin, Huang, Xiaoli, Yang, Shiyong, Zhang, Mingwang, and Yan, Taiming
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- 2022
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5. Circular RNA expression profiles and CircSnd1-miR-135b/c-foxl2 axis analysis in gonadal differentiation of protogynous hermaphroditic ricefield eel Monopterus albus
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He, Zhi, Ma, Zhijun, Yang, Deying, Chen, Qiqi, He, Zhide, Hu, Jiaxiang, Deng, Faqiang, Zhang, Qian, He, Jiayang, Ye, Lijuan, Chen, Hongjun, He, Liang, Huang, Xiaoli, Luo, Wei, Yang, Shiyong, Gu, Xiaobin, Zhang, Mingwang, and Yan, Taiming
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- 2022
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6. Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits
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Xiao, Jie, Zheng, Ruoyu, Bai, Xin, Pu, Jiayan, Chen, Hao, Gu, Xiaobin, Xie, Yue, He, Ran, Xu, Jing, Jing, Bo, Peng, Xuerong, and Yang, Guangyou
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- 2022
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7. Regulatory effects of a novel cysteine protease inhibitor in Baylisascaris schroederi migratory larvae on mice immune cells
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Xu, Jing-Yun, Gu, XiaoBin, Xie, Yue, He, Ran, Xu, Jing, Xiong, Lang, Peng, XueRong, and Yang, GuangYou
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- 2022
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8. Molecular characterization and expression analysis of annexin B3 and B38 as secretory proteins in Echinococcus granulosus
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Song, Hongyu, He, Xue, Du, Xiaodi, Hua, Ruiqi, Xu, Jing, He, Ran, Xie, Yue, Gu, Xiaobin, Peng, Xuerong, and Yang, Guangyou
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- 2021
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9. Comparative analysis of the allergenic characteristics and serodiagnostic potential of recombinant chitinase-like protein-5 and -12 from Sarcoptes scabiei
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Shen, Nengxing, Chen, Yuhang, Wei, Wenrui, Xiong, Lang, Tao, Yuanyuan, Xiao, Jie, Liu, Song, He, Xue, Du, Xiaodi, Gu, Xiaobin, Xie, Yue, Xu, Jing, Peng, Xuerong, and Yang, Guangyou
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- 2021
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10. Molecular characterization and serodiagnostic potential of two serpin proteins in Psoroptes ovis var. cuniculi
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Gu, Xiaobin, Chen, Yuhang, Zhang, Chongyang, Xie, Yue, Shen, Nengxing, Wang, Ce, Zhou, Xuan, Yang, Guangyou, He, Ran, Peng, Xuerong, Yang, Deying, He, Zhi, and Zhong, Zhijun
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- 2020
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11. Molecular characterization of ascaridoid parasites from captive wild carnivores in China using ribosomal and mitochondrial sequences
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Xie, Yue, Li, Yingxin, Gu, Xiaobin, Liu, Yunjian, Zhou, Xuan, Wang, Lu, He, Ran, Peng, Xuerong, and Yang, Guangyou
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- 2020
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12. Pattern and risk factors of local recurrence after nephroureterectomy for upper tract urothelial carcinoma
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Li, Xiaoying, Cui, Ming, Gu, Xiaobin, Fang, Dong, Li, Hongzhen, Qin, Shangbin, Yang, Kunlin, Zhu, Tianzhao, Li, Xuesong, Zhou, Liqun, Gao, Xian-Shu, and Wang, Dian
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- 2020
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13. A pooled analysis of the prognostic value of PD-L1 in melanoma: evidence from 1062 patients
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Yang, Jing, Dong, Meilian, Shui, Yifang, Zhang, Yue, Zhang, Zhigang, Mi, Yin, Zuo, Xiaoxiao, Jiang, Li, Liu, Ke, Liu, Zheyan, Gu, Xiaobin, and Shi, Yonggang
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- 2020
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14. First report on aberrant Ascaris suum infection in a dog, China
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Xie, Yue, Liu, Yunjian, Gu, Xiaobin, Zhou, Xuan, Peng, Xuerong, He, Ran, Guo, Hongrui, Zuo, Zhicai, and Yang, Guangyou
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- 2020
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15. Comparative analysis of host resistance to Sarcoptes scabiei var. cuniculi in two different rabbit breeds
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Wei, Wenrui, Ren, Yongjun, Shen, Nengxing, Song, Hongyu, Xu, Jing, Hua, Ruiqi, Zhang, Haojie, Angel, Christiana, Gu, Xiaobin, Kuang, Liangde, Xie, Yue, Peng, Xuerong, Xie, Xiaohong, and Yang, Guangyou
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- 2019
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16. Elevated PD-L1 expression predicts poor survival outcomes in patients with cervical cancer
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Gu, Xiaobin, Dong, Meilian, Liu, Zheyan, Mi, Yin, Yang, Jing, Zhang, Zhigang, Liu, Ke, Jiang, Li, Zhang, Yue, Dong, Shiliang, and Shi, Yonggang
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- 2019
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17. Molecular characterisation and expression analysis of two heat-shock proteins in Taenia multiceps
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Liu, Yuchen, Guo, Cheng, Dong, Xiaowei, Gu, Xiaobin, Xie, Yue, Lai, Weimin, Peng, Xuerong, and Yang, Guangyou
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- 2019
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18. A chitinase-like protein from Sarcoptes scabiei as a candidate anti-mite vaccine that contributes to immune protection in rabbits
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Shen, Nengxing, Zhang, Haojie, Ren, Yongjun, He, Ran, Xu, Jing, Li, Chunyan, Lai, Weimin, Gu, Xiaobin, Xie, Yue, Peng, Xuerong, and Yang, Guangyou
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- 2018
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19. Genetic characterisation and phylogenetic status of whipworms (Trichuris spp.) from captive non-human primates in China, determined by nuclear and mitochondrial sequencing
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Xie, Yue, Zhao, Bo, Hoberg, Eric P., Li, Mei, Zhou, Xuan, Gu, Xiaobin, Lai, Weimin, Peng, Xuerong, and Yang, Guangyou
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- 2018
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20. Global transcriptome landscape of the rabbit protozoan parasite Eimeria stiedae.
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Xie, Yue, Xiao, Jie, Zhou, Xuan, Gu, Xiaobin, He, Ran, Xu, Jing, Jing, Bo, Peng, Xuerong, and Yang, Guangyou
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EIMERIA ,TRANSCRIPTOMES ,MEROZOITES ,RABBIT diseases ,RNA sequencing ,PROTOZOA ,BROOD parasitism - Abstract
Background: Coccidiosis caused by Eimeria stiedae is a widespread and economically significant disease of rabbits. The lack of studies on the life-cycle development and host interactions of E. stiedae at the molecular level has hampered our understanding of its pathogenesis. Methods: In this study, we present a comprehensive transcriptome landscape of E. stiedae to illustrate its dynamic development from unsporulated oocysts to sporulated oocysts, merozoites, and gametocytes, and to identify genes related to parasite-host interactions during parasitism using combined PacBio single-molecule real-time and Illumina RNA sequencing followed by bioinformatics analysis and qRT-PCR validation. Results: In total, 12,582 non-redundant full-length transcripts were generated with an average length of 1808 bp from the life-cycle stages of E. stiedae. Pairwise comparisons between stages revealed 8775 differentially expressed genes (DEGs) showing highly significant description changes, which compiled a snapshot of the mechanisms underlining asexual and sexual biology of E. stiedae including oocyst sporulation between unsporulated and sporulated oocysts; merozoite replication between sporulated oocysts and merozoites; and gametophyte development and gamete generation between merozoites and gametocytes. Further, 248 DEGs were grouped into nine series clusters and five groups by expression patterns, and showed that parasite–host interaction-related genes predominated in merozoites and gametocytes and were mostly involved in steroid biosynthesis and lipid metabolism and carboxylic acid. Additionally, co-expression analyses identified genes associated with development and host invasion in unsporulated and sporulated oocysts and immune interactions during gametocyte parasitism. Conclusions: This is the first study, to our knowledge, to use the global transcriptome profiles to decipher molecular changes across the E. stiedae life cycle, and these results not only provide important information for the molecular characterization of E. stiedae, but also offer valuable resources to study other apicomplexan parasites with veterinary and public significance. [ABSTRACT FROM AUTHOR]
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- 2021
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21. Characterization of Sarcoptes scabiei cofilin gene and assessment of recombinant cofilin protein as an antigen in indirect-ELISA for diagnosis.
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Yu Zheng, Ran He, Manli He, Xiaobin Gu, Tao Wang, Weimin Lai, Xuerong Peng, Guangyou Yang, Zheng, Yu, He, Ran, He, Manli, Gu, Xiaobin, Wang, Tao, Lai, Weimin, Peng, Xuerong, and Yang, Guangyou
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SARCOPTES scabiei ,RECOMBINANT proteins ,ANTIGENS ,MOLECULAR cloning ,ESCHERICHIA coli ,ENZYME-linked immunosorbent assay ,WESTERN immunoblotting ,LABORATORY rabbits ,ALLERGENS ,ANIMAL experimentation ,GENETIC techniques ,MICROFILAMENT proteins ,RABBITS ,SCABIES - Abstract
Background: Scabies impairs the health of humans and animals and causes heavy economic losses. Traditional diagnostic methods for scabies are inefficient and ineffective, and so far there is no commercial immunodiagnostic or molecular based test for scabies.Methods: Here, we used recombinant Sarcoptes scabiei cofilin protein as an antigen to establish indirect ELISA. S. scabiei cofilin is highly homologous to Dermatophagoides farinae Der f 31 allergen (90% identity). The S. scabiei cofilin gene was cloned and expressed in Escherichia coli to obtain recombinant protein. Western blotting and fluorescence immunohistochemistry were carried out, and we established an indirect ELISA method and detected 33 serum samples from scabies infected rabbits and 30 serum samples from naïve rabbits.Results: Western blotting demonstrated that S. scabiei cofilin possessed good immunogenicity and fluorescence immunohistochemistry showed the S. scabiei cofilin is widespread in the splanchnic area of mites. In ELISA, a cut-off value of 0.188 was determined to judge experimental positive and negative serum values. Specificity and sensitivity of the ELISA were 87.9 and 83.33%, respectively.Conclusions: Recombinant S. scabiei cofilin showed potential value as a diagnostic antigen. The ELISA method established could be used in clinical diagnosis and provide experimental information in minimal or asymptomatic infection. [ABSTRACT FROM AUTHOR]- Published
- 2016
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22. Characterisation and analysis of thioredoxin peroxidase as a potential antigen for the serodiagnosis of sarcoptic mange in rabbits by dot-ELISA.
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Zhang, Runhui, Zheng, Wanpeng, Wu, Xuhang, Jise, Quwu, Ren, Yongjun, Nong, Xiang, Gu, Xiaobin, Wang, Shuxian, Peng, Xuerong, Lai, Songjia, and Yang, Guangyou
- Abstract
Background: Scabies caused by Sarcoptes scabiei is a widespread but a neglected tropical zoonosis. In this study, we characterised a S. scabiei thioredoxin peroxidase (SsTPx) and evaluated a recombinant SsTPx as a diagnostic antigen in rabbits.Methods: The open reading frame of the gene encoding SsTPx-2 was amplified and the recombinant protein was expressed in Escherichia coli cells and purified. SsTPx was localized in mite tissue by immunolocalisation using the purified recombinant protein. Serodiagnosis assays were carried out in 203 New Zealand White rabbit serum samples by dot-ELISA.Result: The open reading frame (489 bp) of the gene encodes an 18.11 kDa protein, which showed highly homology to that of Psoroptes cuniculi (98.77% identity) and belongs to the 2-Cys family of peroxiredoxins. SsTPx was mainly distributed in muscle tissues of mites, integument of the epidermis and the anterior end of S. scabiei. Although SsTPx cross-reactivity with psoroptic mites was observed, the SsTPx dot-ELISA showed excellent diagnostic ability, with 95.3% sensitivity and 93.8% specificity in mange-infected and uninfected groups.Conclusions: This study showed that the purified SsTPx is a highly sensitive antigen for the diagnosis of mange infection by dot-ELISA. This technique is a rapid and convenient method that can be used worldwide for the clinical diagnosis of sarcoptic mange in rabbits, and is especially useful in developing regions. [ABSTRACT FROM AUTHOR]- Published
- 2013
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23. Molecular and biochemical characterization of calmodulin from Echinococcus granulosus.
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Wang N, Zhong X, Song X, Gu X, Lai W, Xie Y, Peng X, and Yang G
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- Amino Acid Sequence, Animals, Calcium chemistry, Calmodulin chemistry, DNA, Complementary genetics, Gene Expression Regulation drug effects, Gene Expression Regulation physiology, Hydrogen Peroxide toxicity, Phylogeny, Protein Binding, RNA, Messenger genetics, RNA, Messenger metabolism, Calmodulin metabolism, Echinococcus granulosus metabolism
- Abstract
Background: Echinococcus granulosus is a harmful cestode parasite that causes cystic echinococcosis in humans as well as various livestock species and wild animals. Calmodulin (CaM), a Ca
2+ sensor protein, is widely expressed in eukaryotes and mediates a variety of cellular signaling activities., Methods: In the present study, the cDNA encoding CaM in Echinococcus granulosus (rEgCaM) was successfully cloned and the molecular and biochemical characterizations carried out. The antigenicity and immunoreactivity of rEgCaM was detected and the preliminary enzyme-linked immunosorbent assay (ELISA)-based serodiagnostic potential of EgCaM was assessed. The locations of this protein in the adult worm and larval stage, and the mRNA expression in different states of E. granulosus protoscoleces (PSCs) were defined clearly. Moreover, the Ca2+ -binding properties of EgCaM were measured., Results: rEgCaM is a highly conserved calcium-binding protein, consisting of 149 amino acids. Immunoblotting analysis revealed that rEgCaM could be identified using E. granulosus infected sheep serum. The use of rEgCaM as an antigen was evaluated by indirect ELISA which exhibited a high sensitivity (90.3%), but low specificity (47.1%). rEgCaM was ubiquitously expressed in protoscoleces and adults of E. granulosus, as well as in the germinal layer of the cyst wall. The mRNA expression level of rEgCaM was increased from the start of H2 O2 exposure and then gradually decreased because of the increased apoptosis of PSCs. In electrophoretic mobility tests and 1-anilinonaphthalene-8-sulfonic acid assays, rEgCaM showed a typical characteristic of a calcium-binding protein., Conclusions: To our knowledge, this is the first report on CaM from E. granulosus and rEgCaM is likely to be involved in some important biological function of E. granulosus as a calcium-binding protein.- Published
- 2017
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24. Analysis of transcriptome data reveals multifactor constraint on codon usage in Taenia multiceps.
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Huang X, Xu J, Chen L, Wang Y, Gu X, Peng X, and Yang G
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- Animals, Evolution, Molecular, Genomics, Helminth Proteins chemistry, Helminth Proteins genetics, Helminth Proteins metabolism, Humans, Hydrophobic and Hydrophilic Interactions, RNA, Messenger genetics, RNA, Messenger metabolism, Taenia metabolism, Codon genetics, Gene Expression Profiling, Taenia genetics
- Abstract
Background: Codon usage bias (CUB) is an important evolutionary feature in genomes that has been widely observed in many organisms. However, the synonymous codon usage pattern in the genome of T. multiceps remains to be clarified. In this study, we analyzed the codon usage of T. multiceps based on the transcriptome data to reveal the constraint factors and to gain an improved understanding of the mechanisms that shape synonymous CUB., Results: Analysis of a total of 8,620 annotated mRNA sequences from T. multiceps indicated only a weak codon bias, with mean GC and GC3 content values of 49.29% and 51.43%, respectively. Our analysis indicated that nucleotide composition, mutational pressure, natural selection, gene expression level, amino acids with grand average of hydropathicity (GRAVY) and aromaticity (Aromo) and the effective selection of amino-acids all contributed to the codon usage in T. multiceps. Among these factors, natural selection was implicated as the major factor affecting the codon usage variation in T. multiceps. The codon usage of ribosome genes was affected mainly by mutations, while the essential genes were affected mainly by selection. In addition, 21codons were identified as "optimal codons". Overall, the optimal codons were GC-rich (GC:AU, 41:22), and ended with G or C (except CGU). Furthermore, different degrees of variation in codon usage were found between T. multiceps and Escherichia coli, yeast, Homo sapiens. However, little difference was found between T. multiceps and Taenia pisiformis., Conclusions: In this study, the codon usage pattern of T. multiceps was analyzed systematically and factors affected CUB were also identified. This is the first study of codon biology in T. multiceps. Understanding the codon usage pattern in T. multiceps can be helpful for the discovery of new genes, molecular genetic engineering and evolutionary studies.
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- 2017
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25. GP50 as a promising early diagnostic antigen for Taenia multiceps infection in goats by indirect ELISA.
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Huang X, Xu J, Wang Y, Guo C, Chen L, Gu X, Lai W, Peng X, and Yang G
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- Animals, Goat Diseases parasitology, Goats, Taeniasis diagnosis, Taeniasis parasitology, Antibodies, Helminth blood, Antigens, Helminth immunology, Enzyme-Linked Immunosorbent Assay methods, Goat Diseases diagnosis, Serologic Tests methods, Taenia immunology, Taeniasis veterinary
- Abstract
Background: Coenurosis is caused by coenurus, the metacestode of Taenia multiceps, which mainly parasitizes the brain and spinal cord of cattle, sheep and goats. To date, no widely-approved methods are available to identify early coenurus infection., Methods: In this study, we identified a full-length cDNA that encodes GP50 (TmGP50) from the transcriptome of T. multiceps, and then cloned and expressed in E. coli. The native proteins in adult stage and coenurus were located via immunofluorescence assays, while the potential of recombinant TmGP50 protein (rTmGP50) for indirect ELISA-based serodiagnostics was assessed using native goat sera. In addition, we orally infected 20 goats with mature T. multiceps eggs. Praziquantel (10%) was given to 10 of the goats 45 days post-infection (p.i.). Blood samples were collected for 17 weeks p.i. from the 20 goats and anti-rTmGP50 antibodies were evaluated using the indirect ELISA established here., Results: The TmGP50 contains an 897 bp open reading frame, in which signal sequence resides in 1 ~ 48 sites and mature polypeptide consists of 282 amino acid residues. Immunofluorescence staining showed that native TmGP50 was localized to the microthrix and parenchymatous zone of the adult parasite and coenurus, and the coenurus cystic wall. The indirect ELISA based on rTmGP50 exhibited a sensitivity of 95.0% and a specificity of 92.6% when detecting GP50 antibodies in sera of naturally infected goats and sheep. In goats experimentally infected with T. multiceps, anti-TmGP50 antibody was detectable from 2 to 17 weeks p.i. in the control group, while the antibody fell below the cut-off value about 3 weeks after praziquantel treatment., Conclusion: Our results indicate that recombinant TmGP50 is a suitable early diagnostic antigen for coenurus infection in goats.
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- 2016
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26. Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus.
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Song X, Yan M, Hu D, Wang Y, Wang N, Gu X, Peng X, and Yang G
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- Amino Acid Sequence, Animals, Echinococcosis blood, Echinococcosis diagnosis, Echinococcosis veterinary, Gene Expression Regulation, Enzymologic physiology, Immunohistochemistry, Larva, Models, Molecular, Protein Conformation, Reproducibility of Results, Sensitivity and Specificity, Serologic Tests, Sheep, Sheep Diseases blood, Sheep Diseases diagnosis, Sheep Diseases parasitology, Cloning, Molecular, Echinococcus granulosus enzymology, Echinococcus granulosus genetics, Enzyme-Linked Immunosorbent Assay methods, Glutaredoxins genetics, Glutaredoxins metabolism
- Abstract
Background: The larval stage of Echinococcus granulosus is the etiological agent of cystic echinococcosis (CE), which causes serious morbidity and mortality in many areas. There is no reliable method to monitor sheep CE. Here, we characterize E. granulosus glutaredoxin 1 (Eg-Grx1) and report an improved immunodiagnostic method for CE., Methods: We cloned and expressed recombinant Eg-Grx1 and generated antibodies. We analyzed the location of the protein in different parasite stages by fluorescence immunohistochemistry, detected the immunogenicity of recombinant Eg-Grx1, and developed an indirect ELISA (iELISA) for CE serodiagnosis., Results: Eg-Grx1 is a classic dithiol Grx with several GSH-binding motifs. Native Eg-Grx1 protein was distributed in the tegument of protoscoleces, the whole germinal layer, and the parenchymatous tissue of adult worms. Recombinant Eg-Grx1 exhibited good immunoreactivity to CE-infected sheep serum. An iELISA using this antigen showed specificity of 64.3 % (9/14) and sensitivity of 1:3200, and the diagnostic accordance rate was 97.9 % (47/48) compared with the results of necropsy., Conclusion: We characterized a novel Grx (Eg-Grx1) from a parasitic helminth and present a comprehensive analysis of the sequence and structure of this protein. The recombinant Eg-Grx1 protein showed good potential serodiagnostic performance, and we established an iELISA method, which may contribute to the surveillance of sheep CE in epidemic areas.
- Published
- 2016
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27. Characterization of Sarcoptes scabiei cofilin gene and assessment of recombinant cofilin protein as an antigen in indirect-ELISA for diagnosis.
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Zheng Y, He R, He M, Gu X, Wang T, Lai W, Peng X, and Yang G
- Subjects
- Actin Depolymerizing Factors immunology, Allergens immunology, Animals, Antigens immunology, Blotting, Western, Cloning, Molecular, Escherichia coli, Rabbits, Recombinant Proteins immunology, Actin Depolymerizing Factors genetics, Enzyme-Linked Immunosorbent Assay methods, Sarcoptes scabiei genetics
- Abstract
Background: Scabies impairs the health of humans and animals and causes heavy economic losses. Traditional diagnostic methods for scabies are inefficient and ineffective, and so far there is no commercial immunodiagnostic or molecular based test for scabies., Methods: Here, we used recombinant Sarcoptes scabiei cofilin protein as an antigen to establish indirect ELISA. S. scabiei cofilin is highly homologous to Dermatophagoides farinae Der f 31 allergen (90% identity). The S. scabiei cofilin gene was cloned and expressed in Escherichia coli to obtain recombinant protein. Western blotting and fluorescence immunohistochemistry were carried out, and we established an indirect ELISA method and detected 33 serum samples from scabies infected rabbits and 30 serum samples from naïve rabbits., Results: Western blotting demonstrated that S. scabiei cofilin possessed good immunogenicity and fluorescence immunohistochemistry showed the S. scabiei cofilin is widespread in the splanchnic area of mites. In ELISA, a cut-off value of 0.188 was determined to judge experimental positive and negative serum values. Specificity and sensitivity of the ELISA were 87.9 and 83.33%, respectively., Conclusions: Recombinant S. scabiei cofilin showed potential value as a diagnostic antigen. The ELISA method established could be used in clinical diagnosis and provide experimental information in minimal or asymptomatic infection.
- Published
- 2016
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28. Expression, tissue localization and serodiagnostic potential of Taenia multiceps acidic ribosomal protein P2.
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Huang X, Chen L, Yang Y, Gu X, Wang Y, Lai W, Peng X, and Yang G
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- Animals, Antigens, Helminth genetics, Antigens, Helminth immunology, Enzyme-Linked Immunosorbent Assay methods, Goat Diseases immunology, Goats, Phosphoproteins genetics, Phosphoproteins immunology, Recombinant Proteins genetics, Recombinant Proteins immunology, Ribosomal Proteins genetics, Ribosomal Proteins immunology, Sensitivity and Specificity, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Taeniasis diagnosis, Taeniasis immunology, Veterinary Medicine methods, Antibodies, Helminth blood, Antigens, Helminth analysis, Clinical Laboratory Techniques methods, Goat Diseases diagnosis, Phosphoproteins analysis, Ribosomal Proteins analysis, Taenia genetics, Taeniasis veterinary
- Abstract
Background: The larval stage of Taenia multiceps, also known as coenurus, is the causative agent of coenurosis, which results in severe health problems in sheep, goats, cattle and other animals that negatively impact on animal husbandry. There is no reliable method to identify coenurus infected goats in the early period of infection., Methods: We identified a full-length cDNA that encodes acidic ribosomal protein P2 from the transcriptome of T. multiceps (TmP2). Following cloning, sequencing and structural analyses were performed using bioinformatics tools. Recombinant TmP2 (rTmP2) was prokaryotically expressed and then used to test immunoreactivity and immunogenicity in immunoblotting assays. The native proteins in adult stage and coenurus were located via immunofluorescence assays, while the potential of rTmP2 for indirect ELISA-based serodiagnostics was assessed using native goat sera. In addition, 20 goats were randomly divided into a drug treatment group and a control group. Each goat was orally given mature, viable T. multiceps eggs. The drug treatment group was given 10% praziquantel by intramuscular injection 45 days post-infection (p.i), and all goats were screened for anti-TmP2 antibodies with the indirect ELISA method established here, once a week for 17 weeks p.i., Results: The open reading frame (366 bp) of the target gene encodes a 12.62 kDa protein, which showed high homology to that from Taenia solium (93% identity) and lacked a signal peptide. Immunofluorescence staining showed that TmP2 was highly localized to the parenchymatous zone of both the adult parasite and the coenurus; besides, it was widely distributed in cystic wall of coenurus. Building on good immunogenic properties, rTmP2-based ELISA exhibited a sensitivity of 95.0% (19/20) and a specificity of 96.3% (26/27) in detecting anti-P2 antibodies in the sera of naturally infected goats and sheep. In goats experimentally infected with T. multiceps, anti-TmP2 antibody was detectable in the control group from 3 to 10 weeks and 15 to 17 weeks p.i. In the drug-treated group, the anti-TmP2 antibody dropped below the cut-off value about 2 weeks after treatment with praziquantel and remained below this critical value until the end of the experiment., Conclusion: The indirect ELISA method developed in this study has the potential for detection of T. multiceps infections in hosts.
- Published
- 2015
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29. Molecular insights into a tetraspanin in the hydatid tapeworm Echinococcus granulosus.
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Hu D, Song X, Xie Y, Zhong X, Wang N, Zheng Y, Gu X, Wang T, Peng X, and Yang G
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- Amino Acid Sequence, Animals, Dogs, Echinococcosis immunology, Echinococcus granulosus classification, Echinococcus granulosus growth & development, Echinococcus granulosus metabolism, Female, Helminth Proteins chemistry, Helminth Proteins metabolism, Humans, Interferon-gamma genetics, Interferon-gamma immunology, Interleukin-12 genetics, Interleukin-12 immunology, Male, Mice, Mice, Inbred ICR, Molecular Sequence Data, Rabbits, Sequence Alignment, Tetraspanins chemistry, Tetraspanins metabolism, Echinococcosis parasitology, Echinococcus granulosus genetics, Helminth Proteins genetics, Tetraspanins genetics
- Abstract
Background: Cystic echinococcosis (hydatid disease), caused by the tapeworm Echinococcus granulosus (class Cestoda; family Taeniidae), is a neglected tropical disease that results in morbidity and mortality in millions of humans, as well as in huge economic losses in the livestock industry globally. Proteins from the tetraspanin family in parasites have recently become regarded as crucial molecules in interaction with hosts in parasitism and are therefore suitable for the development of vaccines and diagnostic agents. However, no information is available to date on E. granulosus tetraspanin., Methods: In this study, a uroplakin-I-like tetraspanin (Eg-TSP1) of E. granulosus was cloned and expressed in E. coli. The immunolocalization of Eg-TSP1 in different life stages of E. granulosus was determined using specific polyclonal antibody. The antibody and cytokine profiles of mice that immunized with recombinant Eg-TSP1 (rEg-TSP1) were measured for the immunogenicity analysis of this protein. Additionally, we use RNA interference method to explore the biological function of Eg-TSP1 in larva of E. granulosus., Results: Immunofluorescence analysis showed that endogenous Eg-TSP1 mainly localized in the tegument of larvae and adults. Significantly elevated levels of antibodies IgG1 and IgG2a and of cytokines IFN-γ and IL-12 were observed in the sera of mice after immunization with rEg-TSP1, suggesting a typical T helper (Th)1-mediated immune response elicited by rEg-TSP1. On further probing the role of Eg-TSP1 in E. granulosus by RNA interference, we found that a thinner tegmental distal cytoplasm was induced in protoscoleces treated with siRNA-132 compared to controls., Conclusions: This is the first report characterizing a tetraspanin from the tapeworm E. granulosus. Our results suggest that Eg-TSP1 is associated with biogenesis of the tegument and maintenance of structural integrity of E. granulosus and could therefore be a candidate intervention target for control of hydatid disease.
- Published
- 2015
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30. Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi.
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Xie Y, Zhou X, Chen L, Zhang Z, Wang C, Gu X, Wang T, Peng X, and Yang G
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- Animals, Base Sequence, Cloning, Molecular, Computational Biology, DNA, Complementary genetics, Enzyme-Linked Immunosorbent Assay, Female, Fluorescent Antibody Technique, Gene Expression Profiling, Glutathione Transferase metabolism, Mice, Molecular Sequence Data, Proteomics methods, Sequence Analysis, DNA, Ascaridoidea enzymology, Glutathione Transferase genetics, Ursidae parasitology
- Abstract
Background: Baylisascaris schroederi, an intestinal nematode of the giant panda, is the cause of the often fatal disease, baylisascariasis. Glutathione S-transferases (GSTs) are versatile enzymes that can affect parasite survival and parasite-host interactions and, are therefore, potential targets for the development of diagnostic tests and vaccines., Methods: In this study, we identified a full-length cDNA that encoded a novel, secretory sigma-like GST (Bsc-GSTσ) from a B. schroederi-omic dataset. Following cloning and sequencing, sequence and structural analyses and comparative modeling were performed using online-bioinformatics and proteomics tools. The recombinant Bsc-GSTσ (rBsc-GSTσ) protein was prokaryotically expressed and then used to detect antigenicity and reactivity using immunoblotting assays. In addition, the native protein in female adult B. schroederi was located via immunofluorescence techniques, while the preliminary ELISA-based serodiagnostic potential of rBsc-GSTσ was assessed in native and infected mouse sera., Results: Bsc-GSTσ contained a 621-bp open reading frame that encoded a polypeptide of 206 amino acids with two typical sigma GST domain profiles, including a GST_N_Sigma_like at the N-terminus and a GST_C_Sigma_like at the C-terminus. The presence of an N-terminal signal sequence indicated that Bsc-GSTσ was a secretory protein. Sequence alignment and phylogenetic analyses showed that Bsc-GSTσ was a nematode-specific member of the Sigma class GSTs and shared the closest genetic distance with its homologue in Ascaris suum. Further comparative structure analyses indicated that Bsc-GSTσ possessed the essential structural motifs (e.g., βαβαββα) and the consensus secondary or tertiary structure that is typical for other characterized GSTσs. Immunolocalization revealed strong distributions of native Bsc-GSTσ in the body hypodermis, lateral chords, gut epithelium, gut microvilli, oviduct epithelium, and ovaries of adult female worms, similar to its homologue in A. suum. Building on good immunogenic properties, rBsc-GSTσ-based ELISA exhibited a sensitivity of 79.1% and a specificity of 82.0% to detect anti-B. schroederi IgG antibodies in the sera of experimentally infected mice., Conclusion: This study presents a comprehensive demonstration of sequence and structural-based analysis of a new, secretory sigma-like GST from a nematode, and its good serodiagnostic performance suggests that rBsc-GSTσ has the potential to detect B. schroederi and, therefore, could be used to develop an ELISA-based serological test to diagnose baylisascariasis in giant pandas.
- Published
- 2015
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31. Absence of genetic structure in Baylisascaris schroederi populations, a giant panda parasite, determined by mitochondrial sequencing.
- Author
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Xie Y, Zhou X, Zhang Z, Wang C, Sun Y, Liu T, Gu X, Wang T, Peng X, and Yang G
- Subjects
- Animals, Ascaridida Infections epidemiology, Ascaridida Infections parasitology, Ascaridoidea isolation & purification, Base Sequence, China epidemiology, DNA, Helminth chemistry, DNA, Helminth genetics, DNA, Mitochondrial chemistry, DNA, Mitochondrial genetics, Genetics, Population, Geography, Haplotypes, Mitochondria genetics, Molecular Sequence Data, Phylogeny, Sequence Analysis, DNA veterinary, Ascaridida Infections veterinary, Ascaridoidea genetics, Genetic Variation, Ursidae parasitology
- Abstract
Background: Infection with the parasitic nematode, Baylisascaris schroederi (Ascaridida: Nematoda), is one of the most important causes of death in giant pandas, and was responsible for half of deaths between 2001 and 2005. Mitochondrial (mt) DNA sequences of parasites can unveil their genetic diversity and depict their likely dynamic evolution and therefore may provide insights into parasite survival and responses to host changes, as well as parasite control., Methods: Based on previous studies, the present study further annotated the genetic variability and structure of B. schroederi populations by combining two different mtDNA markers, ATPase subunit 6 (atp6) and cytochrome c oxidase subunit I (cox1). Both sequences were completely amplified and genetically analyzed among 57 B. schroederi isolates, which were individually collected from ten geographical regions located in three important giant panda habitats in China (Minshan, Qionglai and Qinling mountain ranges)., Results: For the DNA dataset, we identified 20 haplotypes of atp6, 24 haplotypes of cox1, and 39 haplotypes of atp6 + cox1. Further haplotype network and phylogenetic analyses demonstrated that B. schroederi populations were predominantly driven by three common haplotypes, atp6 A1, cox1 C10, and atp6 + cox1 H11. However, due to low rates of gene differentiation between the three populations, both the atp6 and cox1 genes appeared not to be significantly associated with geographical divisions. In addition, high gene flow was detected among the B. schroederi populations, consistent with previous studies, suggesting that this parasite may be essentially homogenous across endemic areas. Finally, neutrality tests and mismatch analysis indicated that B. schroederi had undergone earlier demographic expansion., Conclusions: These results confirmed that B. schroederi populations do not follow a pattern of isolation by distance, further revealing the possible existence of physical connections before geographic separation. This study should also contribute to an improved understanding of the population genetics and evolutionary biology of B. schroederi and assist in the control of baylisascariasis in giant pandas.
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- 2014
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32. Potential of recombinant inorganic pyrophosphatase antigen as a new vaccine candidate against Baylisascaris schroederi in mice.
- Author
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Xie Y, Chen S, Yan Y, Zhang Z, Li D, Yu H, Wang C, Nong X, Zhou X, Gu X, Wang S, Peng X, and Yang G
- Subjects
- Amino Acid Sequence, Animals, Antigens, Helminth chemistry, Antigens, Helminth metabolism, Ascaridida Infections immunology, Ascaridida Infections parasitology, Ascaridoidea metabolism, Female, Helminth Proteins chemistry, Helminth Proteins metabolism, Inorganic Pyrophosphatase chemistry, Inorganic Pyrophosphatase metabolism, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Phylogeny, Polymerase Chain Reaction, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Sequence Alignment, Specific Pathogen-Free Organisms, Antigens, Helminth genetics, Ascaridida Infections prevention & control, Ascaridoidea genetics, Ascaridoidea immunology, Helminth Proteins genetics, Inorganic Pyrophosphatase genetics, Vaccines immunology
- Abstract
The intestinal nematode Baylisascaris schroederi is an important cause of death for wild and captive giant pandas. Inorganic pyrophosphatases (PPases) are critical for development and molting in nematode parasites and represent potential targets for vaccination. Here, a new PPase homologue, Bsc-PYP-1, from B. schroederi was identified and characterized, and its potential as a vaccine candidate was evaluated in a mouse challenge model. Sequence alignment of PPases from nematode parasites and other organisms show that Bsc-PYP-1 is a nematode-specific member of the family I soluble PPases. Immunohistochemistry revealed strong localization of native Bsc-PYP-1 to the body wall, gut epithelium, ovary and uterus of adult female worms. Additionally, Bsc-PYP-1 homologues were found in roundworms infecting humans (Ascaris lumbricoides), swine (Ascaris suum) and dogs (Toxocara canis). In two vaccine trials, recombinant Bsc-PYP-1 (rBsc-PYP-1) formulated with Freund complete adjuvant induced significantly high antigen-specific immunoglobulin (Ig)G but no IgE or IgM responses. Analysis of IgG-subclass profiles revealed a greater increase of IgG1 than IgG2a. Splenocytes from rBsc-PYP-1/FCA-immunized mice secreted low levels of T helper (Th)1-type cytokines, interferon-γ and interleukin (IL)-2, while producing significantly high levels of IL-10 and significantly elevated levels of IL-4 (Th2 cytokines) after stimulation with rBsc-PYP-1 in vitro. Finally, vaccinated mice had 69.02-71.15% reductions (in 2 experiments) in larval recovery 7 days post-challenge (dpc) and 80% survival at 80 dpc. These results suggest that Th2-mediated immunity elicited by rBsc-PYP-1 provides protection against B. schroederi, and the findings should contribute to further development of Bsc-PYP-1 as a candidate vaccine against baylisascariasis.
- Published
- 2013
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33. Identification of neglected cestode Taenia multiceps microRNAs by illumina sequencing and bioinformatic analysis.
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Wu X, Fu Y, Yang D, Xie Y, Zhang R, Zheng W, Nie H, Yan N, Wang N, Wang J, Gu X, Wang S, Peng X, and Yang G
- Subjects
- Algorithms, Animals, Base Sequence, Computational Biology, Gene Expression Regulation, Molecular Sequence Data, Nucleic Acid Conformation, Reproducibility of Results, MicroRNAs genetics, Nucleic Acid Amplification Techniques methods, Taenia classification, Taenia genetics
- Abstract
Background: Worldwide, but especially in developing countries, coenurosis of sheep and other livestock is caused by Taenia multiceps larvae, and zoonotic infections occur in humans. Infections frequently lead to host death, resulting in huge socioeconomic losses. MicroRNAs (miRNAs) have important roles in the post-transcriptional regulation of a large number of animal genes by imperfectly binding target mRNAs. To date, there have been no reports of miRNAs in T. multiceps., Results: In this study, we obtained 12.8 million high quality raw reads from adult T. multiceps small RNA library using Illumina sequencing technology. A total of 796 conserved miRNA families (containing 1,006 miRNAs) from 170,888 unique miRNAs were characterized using miRBase (Release 17.0). Here, we selected three conserved miRNA/miRNA* (antisense strand) duplexes at random and amplified their corresponding precursors using a PCR-based method. Furthermore, 20 candidate novel miRNA precursors were verified by genomic PCR. Among these, six corresponding T. multiceps miRNAs are considered specific for Taeniidae because no homologs were found in other species annotated in miRBase. In addition, 181,077 target sites within T. multiceps transcriptome were predicted for 20 candidate newly miRNAs., Conclusions: Our large-scale investigation of miRNAs in adult T. multiceps provides a substantial platform for improving our understanding of the molecular regulation of T. multiceps and other cestodes development.
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- 2013
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34. Identification of Dirofilaria immitis miRNA using illumina deep sequencing.
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Fu Y, Lan J, Wu X, Yang D, Zhang Z, Nie H, Hou R, Zhang R, Zheng W, Xie Y, Yan N, Yang Z, Wang C, Luo L, Liu L, Gu X, Wang S, Peng X, and Yang G
- Subjects
- Animals, Dirofilariasis parasitology, Dog Diseases parasitology, Dogs, Female, High-Throughput Nucleotide Sequencing veterinary, Male, MicroRNAs metabolism, Molecular Sequence Data, Polymerase Chain Reaction veterinary, Sequence Analysis, RNA veterinary, Dirofilaria immitis genetics, MicroRNAs genetics
- Abstract
The heartworm Dirofilaria immitis is the causative agent of cardiopulmonary dirofilariosis in dogs and cats, which also infects a wide range of wild mammals and humans. The complex life cycle of D. immitis with several developmental stages in its invertebrate mosquito vectors and its vertebrate hosts indicates the importance of miRNA in growth and development, and their ability to regulate infection of mammalian hosts. This study identified the miRNA profiles of D. immitis of zoonotic significance by deep sequencing. A total of 1063 conserved miRNA candidates, including 68 anti-sense miRNA (miRNA*) sequences, were predicted by computational methods and could be grouped into 808 miRNA families. A significant bias towards family members, family abundance and sequence nucleotides was observed. Thirteen novel miRNA candidates were predicted by alignment with the Brugia malayi genome. Eleven out of 13 predicted miRNA candidates were verified by using a PCR-based method. Target genes of the novel miRNA candidates were predicted by using the heartworm transcriptome dataset. To our knowledge, this is the first report of miRNA profiles in D. immitis, which will contribute to a better understanding of the complex biology of this zoonotic filarial nematode and the molecular regulation roles of miRNA involved. Our findings may also become a useful resource for small RNA studies in other filarial parasitic nematodes.
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- 2013
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35. Characterization and evaluation of a Sarcoptes scabiei allergen as a candidate vaccine.
- Author
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Zhang R, Jise Q, Zheng W, Ren Y, Nong X, Wu X, Gu X, Wang S, Peng X, Lai S, and Yang G
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- Amino Acid Sequence, Animals, Gene Expression Regulation physiology, Larva, Molecular Sequence Data, Nymph, Phylogeny, Rabbits, Scabies prevention & control, Tropomyosin chemistry, Tropomyosin genetics, Tropomyosin metabolism, Allergens immunology, Sarcoptes scabiei immunology, Scabies veterinary, Tropomyosin immunology, Vaccines immunology
- Abstract
Background: Sarcoptic mange caused by the mite Sarcoptes scabiei is a worldwide disease affecting both humans and animals. Here we report the molecular characterization and evaluation of a recombinant S. scabiei tropomyosin (SsTm) protein in a vaccination trial in rabbits., Methods: The full-length cDNA was cloned in a bacterial pET vector, and the recombinant protein was expressed in BL21 (DE3) cells and purified. Using specific rabbit antiserum, tropomyosin was localized immunohistochemically in mite tissue sections. Vaccination trials with the recombiant SsTm was carried out in New Zealand rabbits., Results: The full-length open reading frame (ORF) of the 852 bp cloned gene from S. scabiei encodes a 32.9 kDa protein. The amino acid sequence showed 98.94%, 97.89% and 98.59% homology to Dermatophagoides farina and Dermatophagoides pteronyssinus group 10 allergens and Psoroptes ovis tropomyosin, respectively. Tropomyosin was localized immunohistochemically in mite tissue sections mainly in the mouthparts, legs and integument of the epidermis. The predicted cross-reactivity of SsTm indicated that it is an allergenic protein. While vaccination with the recombiant SsTm resulted in high levels of specific IgG (P < 0.01), a low IgE antibody response and no significant protection against S. scabiei challenge were observed. After challenge, specific IgG levels remained significantly higher than the control (P < 0.01), while changes of total IgE levels were not significant (P > 0.05). However, the lesion areas in the vaccination group decreased at the end of the experiment compared with controls., Conclusions: Although vaccination with recombinant SsTm did not efficiently control sarcoptic mange in rabbits, the immunogenic properties of tropomyosin suggest it may be developed as a vaccine with alternative adjuvants or delivery methods.
- Published
- 2012
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