Jiaojiao Lin

Yangzhou University, Shanghai, Shanghai Shi, China

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Publications (53)144.37 Total impact

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    ABSTRACT: Abstract Schistosomes infect around 280 million people worldwide. The worms survive in the veins of the final host, where thioredoxin glutathione reductase (TGR) activity helps the parasites to survive in the aerobic environment. In the present study, we synthesized 2 small interfering RNAs (siRNA S1 and S2) targeting the Schistosoma japonicum (Sj) TGR gene, and used them to knockdown the TGR gene. The knockdown effects of the siRNAs on SjTGR and the thioredoxin reductase (TrxR) activity of SjTGR were evaluated in vitro and in mice infected with schistosomes. The results of transfection with the siRNAs via the soaking method in vitro were confirmed by flow cytometry. S2 siRNA at a final concentration of 200 nM partially inhibited the expression of SjTGR at both the transcript and protein levels in vitro. TrxR-activity was lower in worms in the S2 siRNA-treated group compared with the control groups. Further analysis revealed that purified recombinant SjTGR could remove oxygen free radicals but not H2O2 directly, which may explain the incomplete effects of RNA interference on SjTGR. The results of this preliminary study indicate that SjTGR may play an important role in the clearance of oxygen free radicals and protection of S. japonicum parasites against oxidative damage.
    Journal of Parasitology 03/2014; · 1.32 Impact Factor
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    ABSTRACT: Lethal giant larvae (Lgl) are an evolutionarily conserved tumor suppressor present in fungi and animals. It plays an essential role in establishing apical-basal cell polarity, cell proliferation, differentiation, and tissue organization. Here, we report the presence of Lgl gene in the blood fluke Schistosoma japonicum (SjLgl) (GenBank: KF246684). SjLgl protein was mainly distributed in the unique surface tegument structure by immunofluorescence microscopic staining. Using a simple soaking method, a short interfering RNA (siRNA)-based RNA interference approach knocked down the expression of SjLgl in schistosomula in vitro by up to 89.0 %. Moreover, tail vein injection of SjLgl-siRNA into the infected mice reduced SjLgl mRNA levels in vivo by 48.6-85.3 %, depending on the duration of treatments. SjLgl-specific siRNA treatment during the infection in mice significantly altered the surface structure of adult worm, featured by the disappearance or significant reduction of sharp spines on the inner all of oral and ventral suckers. The siRNA also reduced the hatching rates in eggs produced by treated mice by up to 85.3 %. These observations implied that Lgl plays an important role in the development of tegument in schistosomes, and may be explored as a novel target for developing immuno- and/or small molecule-based therapeutics to control and treat the infections caused by schistosome and other flatworms.
    Parasitology Research 10/2013; · 2.85 Impact Factor
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    ABSTRACT: The interplay between sexes is a prerequisite for female growth, reproductive maturation and egg production, and the basis of schistosome pathopoiesis and propagation. The tegument is in direct contact with the host environment and its surface membranes are particularly crucial for schistosome survival in the definitive host. In this study, a streptavidin-biotin affinity purification technique combined with LC-MS/MS was used to analyze putative tegument-exposed proteins in female and male adult Schistosoma japonicum worms. In total, 179 proteins were identified in females and 300 in males, including 119 proteins common to both sexes, and 60 female biased and 181 male biased proteins. Some (e.g., serpin and CD36-like class B scavenger receptor) were involved in host-schistosome interactions, while some (e.g., gynecophoral canal protein) were important in the interplay between sexes. Gene Ontology analysis revealed that proteins involved in protein glycosylation and lysosome were highly expressed in females, while proteins involved in intracellular signal transduction, regulation of actin filament polymerization and proteasome core complex were highly expressed in males. These results might elucidate physiological differences between sexes. Our study provide new insights into schistosome growth and sexual maturity in the final host and permit the screening of vaccine candidates or drug targets for schistosomiasis.
    Journal of Proteome Research 08/2013; · 5.06 Impact Factor
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    ABSTRACT: The schistosomal tegument is a dynamic host-interactive layer. Proteins exposed to the host on the tegumental surface are important for completion of the parasitic lifecycle. Dysferlin is a member of the ferlin family and is involved in plasma membrane repair. Based on the results of a proteomics study of tegument surface proteins of Schistosoma japonicum in our laboratory, dysferlin was identified as a tegumental protein of S. japonicum. The gene encoding S. japonicum dysferlin (SjDF), which codes for several Ca(2+) binding sites, was cloned, expressed in Escherichia coli, and characterized. Western blot analysis revealed that recombinant SjDF had good immunogenicity. Real-time RT-PCR analysis showed that SjDF was upregulated mainly in adult worms and the transcription level in 42-day-old female worms was significantly higher than that in males. Immunofluorescence analysis revealed that SjDF was mainly distributed in the tegument at various developmental stages. Experimental mice were treated with praziquantel and at 35 days post-infection, we noted that damage to the tegument and subtegument worsened and did not recover at 36 h post-treatment in the high-dose group and was accompanied by downregulation of SjDF mRNA, while the damage was less severe and recovered by this time in the low-dose group, and accompanied by upregulation of SjDF. Our results suggested that SjDF is a tegumental protein that may be important in schistosomal development and may participate in the repair process in muscle and tegument, and could present a viable vaccine candidate for schistosomiasis.
    Parasitology International 07/2013; · 2.30 Impact Factor
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    ABSTRACT: Calcium-binding protein is an indispensable protein which performs extensive and important functions in the growth of Schistosoma japonicum. Based on our primary study on tegument surface proteins of S. japonicun, a cDNA encoding a 66 kDa calcium-binding protein of S. japonicum (Chinese strain) was cloned, sequence analysis revealed that it was identical with that of SjIrV1 of Philippines strains S. japonicum. The expression of SjIrV1 were detected by Real-time PCR, using cDNA templates isolated from 7, 14, 21, 28, 35 and 42 days worms and the results revealed that the gene was expressed in all investigated stages, and the mRNA level of SjIrV1 is much higher in 42 d female worms than that in 42 d male worms. The cDNA containing the open reading frame of IrV1 was subcloned into a pET28a (+) vector and transformed into competent Escherichia coli BL21 for expression. The recombinant protein was purified using a Ni-NTA purification system, and confirmed by high performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS). Western blotting analysis showed that recombinant SjIrV1 (rSjIrV1) could be recognized by the S. japonicum infected mouse serum and the mouse serum specific to rSjIrV1, respectively. Immunofluorescence observation exhibited that SjIrV1 was mainly distributed on the tegument of the 35-day adult worms. ELISA test revealed that IgG, IgG1 and IgG2a antibodies are significantly increased in the serum of rSjIrV1 vaccinated mice. The study suggested that rSjIrV1 might play an important role in the development of S. japonicum.
    Sheng wu gong cheng xue bao = Chinese journal of biotechnology 07/2013; 29(7):891-903.
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    ABSTRACT: BACKGROUND: When compared to the murine permissive host of Schistosoma japonicum, Wistar rats are less susceptible to Schistosoma japonicum infection, and are considered to provide a less suitable microenvironment for parasite growth and development. MicroRNAs (miRNAs), are a class of endogenous, non-coding small RNAs, that impose an additional, highly significant, level of gene regulation within eukaryotes. METHODS: To investigate the regulatory mechanisms provided by miRNA in the schistosome-infected rat model, we utilized a miRNA microarray to compare the progression of miRNA expression within different host tissues both before and 10 days after cercarial infection, in order to identify potential miRNAs with roles in responding to a schistosome infection. RESULTS: Among the analysed miRNAs, 16 within the liver, 61 within the spleen and 10 within the lung, were differentially expressed in infected Wistar rats. Further analysis of the differentially expressed miRNAs revealed that many important signal pathways are triggered after infection with S. japonicum in Wistar rats. These include the signal transduction mechanisms associated with the Wnt and MAPK signaling pathways, cellular differentiation, with a particular emphasis on adipocyte and erythroid differentiation. CONCLUSIONS: The results presented here include the identification of specific differentially expressed miRNAs within the liver, lungs and spleen of Wistar rats. These results highlighted the function of host miRNA regulation during an active schistosome infection. Our study provides a better understanding of the regulatory role of miRNA in schistosome infection, and host--parasite interactions in a non-permissive host environment.
    Parasites & Vectors 04/2013; 6(1):120. · 3.25 Impact Factor
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    ABSTRACT: Retinoid X receptor (RXR) is an important member of the nuclear receptor superfamily of ligand-activated transcription factors that are present in all major groups of metazoans. A full-length cDNA encoding RXR, an orthologue of SmRXR1 in platyhelminth Schistosoma japonicum (SjRXR1) was identified and characterized. The SjRXR1 cDNA is 2806bp long, and contains an open reading frame encoding a 745 amino acid protein. The deduced SjRXR1 protein sequence which was aligned with RXR proteins from other species revealed a highly conserved DNA binding domain (DBD) and moderately conserved ligand binding domain (LBD). The gene structure of SjRXR1 was analyzed and showed that it consists of seven exons spanning 18.4 kbp. The relative mRNA expression of SjRXR1 was evaluated in six different S. japonicum developmental stages in the final host (day 7 to 42 post-infection) and showed higher expression at day 21 and 35. In an in vitro study the transcription of SjRXR1 mRNA was shown to increase almost 3-fold and the SjRXR1 protein expression was also upregulated at the 48h time point by treating the S. japonicum with 5.0μM 9-cis-retinoic acid (RA). Flow cytometry analysis demonstrated that the percentage of HeLa cells expressing SjRXR1LBD-Myc fusion protein is approximately 11%. Over-expression of SjRXR1LBD-Myc in HeLa cells may result in the inhibition of innate apoptosis of this cancer cell line induced by 9-cis-RA. Our studies suggested that the retinoid signaling pathways may be conserved in the platyhelminth. The full cDNA sequence of SjRXR1 reported here has been submitted to the GenBank with accession no. JX111997.
    Molecular and Biochemical Parasitology 02/2013; · 2.73 Impact Factor
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    ABSTRACT: Schistosomiasis remains an important global public health problem that affects 200 million people in 76 countries. The molecular mechanisms of host-parasite interaction are complex, and in schistosome infection regulation of microRNA (miRNA) and the host micro-environment may be involved. In this study, an miRNA microarray was applied to investigate differences in miRNA expression in different tissues of mice before and 10 days post infection. In total, 220 miRNAs were detected in different tissues of the BALB/c mice before and after infection, including 8 miRNAs in liver, 8 in spleen and 28 in the lungs with up-regulated expression, and 3 miRNAs in liver, 5 in spleen and 28 in the lungs with down-regulated expression in mice 10 days post infection with schistosomes. The functions of these differentially expressed miRNAs are related mainly to the immune response, nutrient metabolism, cell differentiation, apoptosis, and signal pathways. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the differentially expressed miRNAs revealed that many important biological pathways are triggered by schistosome infection in BALB/c mice, such as the MAPK signaling pathway, insulin signaling pathway, Toll-like receptor signaling pathway and TGF-β signaling pathway.The results reveal that miRNAs may be an important regulator of schistosome-host interaction in the early phase of Schistosoma japonicum infection. The data presented here provide valuable information to increase understanding of the regulatory function of the miRNAs in the host micro-environment, as well as the mechanism of host-parasite interactions. This may be helpful in the search for potential new drugs, and for biomarkers of early S. japonicum infection applicable in the future control of schistosomiasis.
    Molecular and Biochemical Parasitology 02/2013; · 2.73 Impact Factor
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    ABSTRACT: Myoferlin is a member of the ferlin family of proteins, which are involved in plasma membrane repair, and has been identified as one of the tegument proteins of Schistosoma japonicum. The tegument proteins are potential candidates for vaccines and new drug targets. In this study, myoferlin of S. japonicum (SjMF) was cloned, expressed and characterized, the potential of SjMF recombinant protein (rSjMF) as a vaccine candidate was evaluated, and the effect of praziquantel on SjMF was detected by Real-time PCR. Immunofluorescence showed that this protein was mainly distributed on the surface of worms at different stages. Sequence analysis revealed that the SjMF open reading frame was conserved at all stages of the S. japonicum life cycle. And SjMF transcription was upregulated in 42-day-old worms, and was significantly higher in female worms. Western blotting revealed that rSjMF showed strong immunogenicity. The cytokine profile and IgG isotype analysis demonstrated that rSjMF plus ISA206 immunization induced a mixed T helper (Th)1/Th2 response. Purified rSjMF emulsified with ISA206 adjuvant significantly reduced worm burden from 21.8% to 23.21% and liver egg number from42.58% to 28.35%. Besides, SjMF transcription was downregulated when worms were exposed to low-dose praziquantel (PZQ) and upregulated when PZQ was degraded, accompanied by recovery of damaged tegument. When worms were exposed to high-dose PZQ, SjMF transcription was downregulated all the time and the damaged tegument did not recover. These findings indicated that SjMF is a potential vaccine against S. japonicum and provides the basis for further investigations into the biological function of SjMF.
    PLoS ONE 01/2013; 8(6):e66396. · 3.73 Impact Factor
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    ABSTRACT: Schistosomiasis, caused by infection with Schistosoma species, remains an important parasitic zoonosis. Thioredoxin glutathione reductase of Schistosoma japonicum (SjTGR) plays an important role in the development of the parasite and for its survival. Here we present a recombinant plasmid DNA vaccine, pVAX1/SjTGR, to estimate its protection against S. japonicum in BALB/c mice. The DNA vaccine administrated by particle bombardment induced higher protection than by intramuscular injection. All animals vaccinated with pVAX1/SjTGR developed significant specific anti-SjTGR antibodies than control groups. Moreover, animals immunized by gene gun exhibited a splenocyte proliferative response, with an increase in IFN- γ and IL-4. The recombinant plasmid administrated by gene gun achieved a medium protective efficacy of 27.83-38.83% (P < 0.01) of worm reduction and 40.38-44.51% (P < 0.01) of liver egg count reduction. It suggests that different modes of administering a DNA vaccine can influence the protective efficacy induced by the vaccine. Interestingly, from the enzymatic activity results, we found that worms obtained from pVAX1/SjTGR-vaccinated animals expressed lower enzymatic activity than the control group and the antibodies weakened the enzymatic activity of SjTGR in vitro, too. It implies that the high-level antibodies may contribute to the protective effects.
    BioMed research international. 01/2013; 2013:952416.
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    ABSTRACT: Water buffalo and goats are natural hosts for S. japonicum in endemic areas of China. The susceptibility of these two hosts to schistosome infection is different, as water buffalo are less conducive to S. japonicum growth and development. To identify genes that may affect schistosome development and survival, we compared gene expression profiles of schistosomes derived from these two natural hosts using high-throughput microarray technology. The worm recovery rate was lower and the length and width of worms from water buffalo were smaller compared to those from goats following S. japonicum infection for 7 weeks. Besides obvious morphological difference between the schistosomes derived from the two hosts, differences were also observed by scanning and transmission electron microscopy. Microarray analysis showed differentially expressed gene patterns for parasites from the two hosts, which revealed that genes related to lipid and nucleotide metabolism, as well as protein folding, sorting, and degradation were upregulated, while others associated with signal transduction, endocrine function, development, immune function, endocytosis, and amino acid/carbohydrate/glycan metabolism were downregulated in schistosomes from water buffalo. KEGG pathway analysis deduced that the differentially expressed genes mainly involved lipid metabolism, the MAPK and ErbB signaling pathways, progesterone-mediated oocyte maturation, dorso-ventral axis formation, reproduction, and endocytosis, etc. The microarray gene analysis in schistosomes derived from water buffalo and goats provide a useful platform to disclose differences determining S. japonicum host compatibility to better understand the interplay between natural hosts and parasites, and identify schistosome target genes associated with susceptibility to screen vaccine candidates.
    PLoS ONE 01/2013; 8(8):e70367. · 3.73 Impact Factor
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    ABSTRACT: The reed vole Microtus fortis is the only mammal known in China in which the growth, development and maturation of schistosomes (Schistosoma japonicum) is prevented. It might be that the anti-schistosomiasis mechanisms of M. fortis associate with microRNA-mediated gene expression, given that the latter has been found to be involved in gene regulation in eukaryotes. In the present study, the difference between pathological changes in tissues of M. fortis and of mice (Mus musculus) post-schistosome infection were observed by using hematoxylin-eosin staining. In addition, microarray technique was applied to identify differentially expressed miRNAs in the same tissues before and post-infection to analyze the potential roles of miRNAs in schistosome infection in these two different types of host. Histological analyses showed that S. japonicum infection in M. fortis resulted in a more intensive inflammatory response and pathological change than in mice. The microarray analysis revealed that 162 miRNAs were expressed in both species, with 12 in liver, 32 in spleen and 34 in lung being differentially expressed in M. fortis. The functions of the differentially expressed miRNAs were mainly revolved in nutrient metabolism, immune regulation, etc. Further analysis revealed that important signaling pathways were triggered after infection by S. japonicum in M. fortis but not in the mice. These results provide new insights into the general mechanisms of regulation in the non-permissive schistosome host M. fortis that exploits potential miRNA regulatory networks. Such information will help improve current understanding of schistosome development and host-parasite interactions.
    PLoS ONE 01/2013; 8(12):e85080. · 3.73 Impact Factor
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    ABSTRACT: Schistosoma japonicum has a complex lifecycle and exhibits dramatic changes in its biology and morphology at different developmental stages. The schistosomulum and adult worm are two stages of this complex lifecycle and differentially expressed proteins in these two stages should be important for survival, development, and reproduction of the parasites. In this study, soluble and hydrophobic proteins were extracted from eggs, cercariae, schistosomula (8 d and 19 d), and male and female adult worms (42 d) of Schistosoma japonicum, and separated by two-dimensional (2D) gel electrophoresis. A total of 1376±52, 928±61, 1465±41, 1230±30, 904±34, and 1080±26 soluble proteins and 1437±44, 845±53, 986±22, 1145±35, 1066±39, and 1123±45 hydrophobic proteins were separated from eggs, cercariae, schistosomula (8 d and 19 d), and male and female adult worms (42 d), respectively. There were 65±14, 27±7, 37 ±17 and 48±9 soluble protein spots only present in schistosomula (8 days and/or 19 days) and adult schistosomes (male and/or female). We successfully identified 22 spots from schistosomula and 11 spots from adult schistosomes by mass spectrometry. Quantitative real-time RT-PCR was used to examine six differentially expressed proteins at the transcription level. These proteins only found in schistosomula or adults stage by the proteomics analysis were highly expressed in the corresponding stage at mRNA level. Bioinformatics analysis showed that the differentially expressed proteins from schistosomula were mainly involved in cellular metabolic processes, stress response and developmental process. Differentially expressed proteins from adult schistosomes were involved with gene expression and protein metabolism processes. The results of this study might provide new insights to stimulate further exploration of the mechanism of growth and development in schistosomes and help identify candidate molecules for developing new vaccines or drugs.
    Acta tropica 12/2012; · 2.79 Impact Factor
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    ABSTRACT: Protein phosphorylation is an important posttranslational modification in many organisms that regulates numerous cellular processes. However, it remains poorly characterized in schistosomes, causative agent of schistosomiasis in human and related animals. In the present study, we characterized phosphorylated proteins in different stages and sex of Schistosoma japonicum (S. japonicum) including schistosomula (14 d), adult females (35 d) and adult males (35 d) by a titanium dioxide (TiO(2)) based phosphoproteomic method. A total of 180 phosphopeptides were identified in 148 proteins. Our further studies revealed that heat shock protein 90 (Hsp90), one of phosphoproteins co-detected in the different stage and sex of schistosomes, may play an important roles in the regulation of schistosome development by directly or indirectly interacting with other co-detected signal molecules. Additionally, some phosphoproteins were shown to be detected in a gender specific manner and the expressions of these proteins were further validated either by immunohistochemistry or by real time RT-PCR at transcript levels between male and female schistosomes. In summary, these findings as well as the providing of an inventory of phosphoproteins are expected to provide new insights in schistosome development and sexual maturation, and then may result in the development of novel interventions against schistosomiasis.
    Journal of Proteome Research 12/2012; · 5.06 Impact Factor
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    ABSTRACT: Apoptosis is an important aspect of a number of biological processes, from embryogenesis to the stress-injury response. It plays a central role in balancing cell proliferation and tissue remodeling activity in many organisms. In the present study, apoptosis in 14days post infection schistosomula was evaluated using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) assays and DAPI staining. Additionally, flow cytometry using the Annexin V-FITC/propidium iodide (PI) (Annexin V/PI) assay confirmed the percentage of early apoptotic, late apoptotic, and necrotic cells in 14 and 23days post infection worms. Conserved Domain Database (CDD) BLAST analysis and alignment analysis of known schistosome proteins demonstrated the feasibility of detecting the activity of Caspase-3 and -7 using the Caspase3/7 Glo analysis assay. Analysis of Caspase-3 and -7 activity in schistosome demonstrated that both caspases were active in each developmental stage of S. japonicum, but was highest in the 14days post infection schistosomula. Additionally, the caspase peptide inhibitor (Z-VAD-FMK) inhibited the Caspase-3/7 activity at all developmental stages examined. Therefore, we hypothesized that two main signaling pathways are involved in apoptosis in S. japonicum, the caspase cascade and the mitochondrial-initiated pathway. We have constructed a model of these two pathways, including how they may interact and their biological outcomes. qRT-PCR analyses of the gene expression profiles of apoptosis-related genes supported our hypothesis of the relationship between the apoptotic pathway and parasite development. The data presented here demonstrates that apoptosis is an important biological process for the survival and development of the schistosome, and identifies potential novel therapeutic targets.
    Parasitology International 11/2012; · 2.30 Impact Factor
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    ABSTRACT: Glutamine synthetase catalyzes the synthesis of glutamine, providing nitrogen for the production of purines, pyrimidines, amino acids, and other compounds required in many pivotal cellular events. Herein, a full-length cDNA encoding Schistosoma japonicum glutamine synthetase (SjGS) was isolated from 21-day schistosomes. The entire open reading frame of SjGS contains a 1,095-bp coding region corresponding to 364 amino acids with a calculated molecular weight of 40.7 kDa. NCBIP blast shows that the putative amino acid of SjGS contains a classic β-grasp domain and a catalytic domain of glutamine synthetase. The relative mRNA expression of SjGS was evaluated in 7-, 13-, 21-, 28-, 35-, and 42-day worms of S. japonicum in the final host and higher expression at day 21, and 42 worms were observed. This protein was also detected in worm extracts using Western blot. Immunofluorescence studies indicated that the SjGS protein was mainly distributed on tegument and parenchyma in 28-day adult worms. The recombinant glutamine synthetase with a molecular weight of 45 kDa was expressed in Escherichia coli and purified in its active form. The enzyme activity of the recombinant protein was 3.30 ± 0.67 U.μg-1. The enzyme activity was highly stable over a wide range of pH (6-9) and temperature (25-40 °C) under physiological conditions. The transcription of SjGS was upregulated in praziquantel-treated worms at 2-, 4-, and 24-h posttreatment compared with the untreated control. As a first step towards the clarification of the role of glutamine synthetase in schistosome species, we have cloned and characterized cDNAs encoding SjGS in S. japonicum, and the data presented suggest that SjGS is an important molecule in the development of the schistosome.
    Parasitology Research 09/2012; · 2.85 Impact Factor
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    ABSTRACT: Thyroid hormones (TH) modulate growth, development and differentiation and metabolic processes by interacting with thyroid hormone receptors (THRs). The purpose of this study was to identify a novel thyroid hormone receptor beta encoding gene of Schistosoma japonicum (SjTHRβ) and to investigate its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice. The full-length cDNA sequence of SjTHRβ, its gene organization, and its transcript levels were characterized, and the phylogenetic relationship between THR, RAR and RXR from other organisms were analysis, the ability of this protein binding to a conserved DNA core motif, and its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice were evaluated. The SjTHRβ cDNA was cloned, verified by 5' and 3' Rapid Amplification of cDNA Ends and shown to be polyadenylated at the 3'end, suggesting the transcript is full-length. SjTHRβ is homologous to THRs from other species and has a predicted conservative DNA binding domain and ligand binding domain that normally characterizes these receptors. A comparative quantitative PCR analysis showed that SjTHRβ was the highest expressed in 21d worms and the lowest in 7 d and 13 d schistosomula. The cDNA corresponding to DNA binding domain (SjTHRβ-DBD) and ligand binding domain (SjTHRβ-LBD) were cloned and subsequently expressed in E coli. The expressed proteins were used to immunize mice and generate specific serum against recombinant SjTHRβ (rSjTHRβ). Western blotting revealed that anti-rSjTHRβ-LBD serum recognized two protein bands in extracts from 21 d worm with molecular sizes of approximately 95 kDa and 72 kDa. Electrophoretic mobility shift assay (EMSA) analysis showed that rSjTHRβ-DBD could bind to a conserved DNA core motif. Immunization of BALB/c mice with rSjTHRβ-LBD could induce partial protective efficacy(27.52% worm reduction and 29.50% liver eggs reduction)against schistosome infection. Enzyme-linked immunosorbent assay showed that mice vaccinated with recombinant SjTHRβ-LBD (rSjTHRβ-LBD) generated increased levels of specific IgG, IgG1 and IgG2a antibody. Bio-plex analysis demonstrated that rSjTHRβ-LBD induced considerably higher levels of T helper 1 cytokines (IL-2, IL-12 and TNF-α) than T helper 2 cytokines (IL-10, IL-4), suggesting that rSjTHRβ-LBD vaccination could stimulate mixed Th1/Th2 types with Th1 dominant immune responses. Our study presented here identified SjTHRβ as a new schistosome THR that might play an important role in host-parasite interaction and be a vaccine candidate for schistosomiasis.
    Parasites & Vectors 08/2012; 5:172. · 3.25 Impact Factor
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    ABSTRACT: Abstract We analyzed proteins that were differentially expressed by 10-day-old schistosomula from 3 different hosts and determined that a functional thioredoxin peroxidase-2 gene has an important antioxidant role in Schistosoma japonicum, which we investigated further. A full-length cDNA encoding the S. japonicum thioredoxin peroxidase-2 (SjTPx-2) had an open reading frame (ORF) of 681 bp that encoded 227 amino acids with a signal peptide of 24 amino acids. A cDNA encoding SjTPx-2 without the signal peptides sequence was isolated from 42-day-old schistosome cDNAs. Real-time quantitative RT-PCR analysis revealed that SjTPx-2 was upregulated in 7- and 13-day-old schistosomes, while the expression level in females was around 2-fold higher than that in male worms at 42 days. SjTPx was subcloned into pET28a(+) and expressed as both inclusion bodies and supernatant in Escherichia coli BL21 (DE3) cells. Western blotting showed that the recombinant SjTPx-2 (rSjTPx-2) was immunogenic. The purified recombinant protein could form disulfide-bonded dimers and it had peroxidase activity in vitro. An immunoprotection experiment in BALB/c mice showed that vaccination with recombinant SjTPx-2 could induce 31.2% and 34.0% reductions in the numbers of worms and eggs in the liver, respectively. This study suggests that SjTPx-2 may be an important antioxidative enzyme in scavenging ROS and it may be a potential vaccine candidate or new drug target for schistosomiasis.
    Journal of Parasitology 06/2012; · 1.32 Impact Factor
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    ABSTRACT: This work reports the prevention outcomes of a praziquantel (PZQ) implant against the infection of Schistosoma japonicum in mice. The PZQ implant produced stable plasma PZQ concentrations in a range of 100-1300 ng/mL for a period of 70 days, by releasing PZQ in subcutaneous tissues in a sustained manner. To assess the prevention effects, the mice were infected at varying times after implantation. All the mice were sacrificed at 6 weeks after infection for worm and egg recovery and counting, worm morphological examination, determination of egg-hatching rates, and analysis of hepatic histology. The infection was successfully prevented for mice with early infection times (within 2-3 weeks), as nearly no worms, paired worms, eggs, or miracidia were recovered. However, in mice with late infection times (after 3 weeks), the prevention effects were diminished due to the decreased plasma PZQ concentrations at late times. Interestingly, the implants showed robust prevention effects on repeated infection at 1 and 3 weeks. In the infection-prevented mouse livers, no granuloma formation or granulomatous inflammation was observed. The results demonstrated that by blocking the development of infecting miracidia and by deactivating the eggs, the PZQ implants encouragingly prevented the S. japonicum infection and avoided liver damage.
    Experimental Parasitology 06/2012; 131(4):442-7. · 2.15 Impact Factor
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    ABSTRACT: Type V collagen is a component of non-cartilaginous tissues and is important in the determination of fibril structure and matrix organization, although its functions are still poorly understood. In this report, RNA interference (RNAi) approaches were used to investigate the effects of knockdown of the schistosome type V collagen (SjColV) gene. In this study, three different short interfering (si) RNAs targeting different regions of the gene were designed to suppress the expression of SjColV in Schistosoma japonicum using a soaking method. By establishing controls for measuring off-target RNAi effects, we found that different siRNA sequences had different levels of effectiveness. Although all the siRNAs tested reduced SjColV transcript levels, the S1 siRNA consistently reduced SjColV expression to >99 % of the control. In the following experiments, S1 siRNA was adapted to inhibit SjColV expression, and the silencing effects were detected by real-time PCR and Western blot. The spawning and egg hatching of parasites were calculated, while the worms' morphology was taken by scanning electron microscopy. The results show that silencing the expression of SjColV significantly affects the spawning and egg hatching of S. japonicum, and it also affects the worms' morphology.
    Parasitology Research 05/2012; 111(3):1251-7. · 2.85 Impact Factor

Publication Stats

193 Citations
400 Downloads
3k Views
144.37 Total Impact Points

Institutions

  • 2013
    • Yangzhou University
      • College of Veterinary Medicine
      Shanghai, Shanghai Shi, China
    • Chinese Academy of Agricultural Sciences
      • Key Laboratory of Animal Parasitology
      Peping, Beijing, China
  • 2012–2013
    • Xiamen University
      Amoy, Fujian, China
  • 2011–2012
    • Shanghai Jiao Tong University
      • School of Pharmacy
      Shanghai, Shanghai Shi, China
  • 2008–2011
    • Shanghai Veterinary Research Institute
      Shanghai, Shanghai Shi, China
    • Academy of Military Medical Sciences
      T’ien-ching-shih, Tianjin Shi, China