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1.
Article in English | WPRIM | ID: wpr-716525

ABSTRACT

BACKGROUND: Schistosoma haematobium which causes urogenital schistosomiasis (UGS) is highly prevalent in African countries. Urine microscopy (UM) is the first-line diagnostic method of UGS. Enzyme-linked immunosorbent assay (ELISA) is a common method for screening many parasite infections primarily or alternatively. The present study established an in-house diagnostic system by ELISA and evaluated its diagnostic efficacy in comparison with UM for screening UGS in White Nile State, Republic of Sudan, 2011–2013. METHODS: A total of 490 participants were screened by UM or ELISA, and 149 by both. The in-house ELISA system was established employing soluble egg antigen of S. haematobium and the cut-off absorbance was set at 0.270. RESULTS: Of the 149 subjects, 58 participants (38.9%) were positive by UM, 119 (79.9%) were positive by ELISA and 82 (55.0%) showed consistently positive or negative results by both methods. The diagnostic sensitivity of ELISA was 94.8% and specificity was 29.7% based on UM results. The ELISA positive serum samples also cross-reacted with egg antigens of Schistosoma mansoni and Schistosoma japonicum. CONCLUSION: We have established in-house ELISA for screening serum immunoglobulin (Ig) G antibodies by employing soluble egg antigen of S. haematobium for diagnosis of UGS with 94.8% sensitivity and 29.7% specificity. The ELISA system can supplement the conventional diagnosis by UM.


Subject(s)
Antibodies , Diagnosis , Enzyme-Linked Immunosorbent Assay , Immunoglobulins , Mass Screening , Methods , Microscopy , Ovum , Parasites , Schistosoma haematobium , Schistosoma japonicum , Schistosoma mansoni , Schistosoma , Schistosomiasis haematobia , Sensitivity and Specificity , Sudan
2.
Rev. Soc. Bras. Med. Trop ; 50(1): 126-129, Jan.-Feb. 2017. tab
Article in English | LILACS | ID: biblio-1041397

ABSTRACT

Abstract: INTRODUCTION: The activity of garlic oil extract against Schistosoma japonicum cercariae was evaluated. METHODS: The in vitro and in vivo cercaricidal activities against S. japonicum larvae were determined. RESULTS: Exposure to ≥ 10-6 (v/v) garlic emulsions for 30 min led to 100% cercariae mortality; pre-exposure treatment with ≥ 10-4 (v/v) garlic emulsions showed 100% preventive efficacy against S. japonicum infection, while pre-treatment with 10-5 and 10-6 (v/v) emulsions achieved 20%-40% preventive efficacy and 35.2%-63.6% worm burden reduction. CONCLUSIONS: Garlic oil extract has activity against S. japonicum larvae and a promising preventive efficacy against S. japonicum infection.


Subject(s)
Animals , Female , Schistosoma japonicum/drug effects , Plant Extracts/pharmacology , Cercaria/drug effects , Garlic/chemistry , Time Factors , Parasitic Sensitivity Tests , Mice
3.
Article in English | WPRIM | ID: wpr-50086

ABSTRACT

China still has more than 30,000 patients of advanced schistosomiasis while new cases being reported consistently. D-dimer is a fibrin degradation product. As ascites being the dominating symptom in advanced schistosomiasis, the present study aimed to explore a prediction model of ascites with D-dimer and other clinical easy-achievable indicators. A case-control study nested in a prospective cohort was conducted in schistosomiasis-endemic area of southern China. A total of 291 patients of advanced schistosomiasis were first investigated in 2013 and further followed in 2014. Information on clinical history, physical examination, and abdominal ultrasonography, including the symptom of ascites was repeatedly collected. Result showed 44 patients having ascites. Most of the patients' ascites were confined in the kidney area with median area of 20 mm². The level of plasma D-dimer and pertinent liver function indicators were measured at the initial investigation in 2013. Compared with those without ascites, cases with ascites had significantly higher levels of D-dimer (0.71±2.44 μg/L vs 0.48±2.12 μg/L, P=0.005), as well ALB (44.5 vs 46.2, g/L) and Type IV collagen (50.04 vs 44.50 μg/L). Receiver operating characteristic curve analyses indicated a moderate predictive value of D-dimer by its own area under curve (AUC) of 0.64 (95% CI: 0.54–0.73) and the cutoff value as 0.81 μg/L. Dichotomized by the cutoff level, D-dimer along with other categorical variables generated a prediction model with AUC of 0.76 (95% CI: 0.68–0.89). Risks of patients with specific characteristics in the prediction model were summarized. Our study suggests that the plasma D-dimer level is a reliable predictor for incident ascites in advanced schistosomiasis japonica patients.


Subject(s)
Area Under Curve , Ascites , Case-Control Studies , China , Cohort Studies , Collagen Type IV , Fibrin , Humans , Kidney , Liver , Physical Examination , Plasma , Prospective Studies , ROC Curve , Schistosoma japonicum , Schistosomiasis japonica , Schistosomiasis , Ultrasonography
4.
Chinese Journal of Biotechnology ; (12): 889-900, 2016.
Article in Chinese | WPRIM | ID: wpr-242289

ABSTRACT

For further research of the apoptosis mechanism of Schistosoma japonicum (S. japonicum). The cDNA encoding Sjcaspase3 of Schistosoma japonicum was amplified by polymerase chain reaction (PCR) technique, which contained 900 nucleotides and encoded 299 amino acids. The theory molecular weight and isoelectric point (PI) of the deduced protein is 33.5 kDa and 6.39, respectively. Real-time PCR was used to analyze the transcription profiles of Sjcaspase3 at different development stages of S. japonicum. The results showed that this gene was expressed in all stages of S. japonicum with the highest expression in 21d worms, and the level of gene transcription in 42 d female worms was higher than that of male worms. The recombinant plasmid pXJ40-FLAG-Sjcaspase3 was constructed and transfection into Hela cells successfully. Real-time PCR and Western blotting analysis showed Sjcaspase3 was successfully expressed in Hela cells. Enzyme activity analysis revealed that recombinant Sjcaspase3 possessed the activity to cut substrate DEVD. Flow cytometry proved that Sjcaspase3 could induce early apoptosis of Hela cells. The results provide the basis for proceeding further study on the biological function of Sjcaspase3 and better understand the apoptosis mechanism of S. japonicum.


Subject(s)
Animals , Apoptosis , Blotting, Western , Caspase 3 , Genetics , Metabolism , Cloning, Molecular , DNA, Complementary , Female , HeLa Cells , Helminth Proteins , Genetics , Metabolism , Humans , Male , Real-Time Polymerase Chain Reaction , Recombinant Proteins , Schistosoma japonicum
5.
Article in English | WPRIM | ID: wpr-207497

ABSTRACT

Whilst archaeological evidence for many aspects of life in ancient China is well studied, there has been much less interest in ancient infectious diseases, such as intestinal parasites in past Chinese populations. Here, we bring together evidence from mummies, ancient latrines, and pelvic soil from burials, dating from the Neolithic Period to the Qing Dynasty, in order to better understand the health of the past inhabitants of China and the diseases endemic in the region. Seven species of intestinal parasite have been identified, namely roundworm, whipworm, Chinese liver fluke, oriental schistosome, pinworm, Taenia sp. tapeworm, and the intestinal fluke Fasciolopsis buski. It was found that in the past, roundworm, whipworm, and Chinese liver fluke appear to have been much more common than the other species. While roundworm and whipworm remained common into the late 20th century, Chinese liver fluke seems to have undergone a marked decline in its prevalence over time. The iconic transport route known as the Silk Road has been shown to have acted as a vector for the transmission of ancient diseases, highlighted by the discovery of Chinese liver fluke in a 2,000 year-old relay station in northwest China, 1,500 km outside its endemic range.


Subject(s)
Archaeology , Asians , Burial , Cestoda , China , Clonorchis sinensis , Communicable Diseases , Enterobius , Fasciola hepatica , Fasciolidae , Helminths , Humans , Mummies , Parasites , Prevalence , Schistosoma japonicum , Silk , Soil , Taenia , Toilet Facilities , Trematoda
6.
Article in English | WPRIM | ID: wpr-285260

ABSTRACT

Infection of schistosomiasis japonica may eventually lead to liver fibrosis, and no effective antifibrotic therapies are available but liver transplantation. Hedgehog (HH) signaling pathway has been involved in the process and is a promising target for treating liver fibrosis. This study aimed to explore the effects of pentoxifylline (PTX) on liver fibrosis induced by schistosoma japonicum infection by inhibiting the HH signaling pathway. Phorbol12-myristate13-acetate (PMA) was used to induce human acute mononuclear leukemia cells THP-1 to differentiate into macrophages. The THP-1-derived macrophages were stimulated by soluble egg antigen (SEA), and the culture supernatants were collected for detection of activation of macrophages. Cell Counting Kit-8 (CCK-8) was used to detect the cytotoxicity of the culture supernatant and PTX on the LX-2 cells. The LX-2 cells were administered with activated culture supernatant from macrophages and(or) PTX to detect the transforming growth factor-β gene expression. The mRNA expression of shh and gli-1, key parts in HH signaling pathway, was detected. The mRNA expression of shh and gli-1 was increased in LX-2 cells treated with activated macrophages-derived culture supernatant, suggesting HH signaling pathway may play a key role in the activation process of hepatic stellate cells (HSCs). The expression of these genes decreased in LX-2 cells co-cultured with both activated macrophages-derived culture supernatant and PTX, indicating PTX could suppress the activation process of HSCs. In conclusion, these data provide evidence that PTX prevents liver fibrogenesis in vitro by the suppression of HH signaling pathway.


Subject(s)
Animals , Antigens, Helminth , Pharmacology , Cell Culture Techniques , Cell Differentiation , Cell Line , Culture Media, Conditioned , Chemistry , Pharmacology , Gene Expression Regulation , Hedgehog Proteins , Genetics , Allergy and Immunology , Hepatic Stellate Cells , Cell Biology , Metabolism , Humans , Liver Cirrhosis , Metabolism , Parasitology , Macrophage Activation , Macrophages , Cell Biology , Allergy and Immunology , Models, Biological , Monocytes , Cell Biology , Metabolism , Pentoxifylline , Pharmacology , Phosphodiesterase Inhibitors , Pharmacology , RNA, Messenger , Genetics , Allergy and Immunology , Schistosoma japonicum , Chemistry , Signal Transduction , Tetradecanoylphorbol Acetate , Pharmacology , Zinc Finger Protein GLI1 , Genetics , Allergy and Immunology , Zygote , Chemistry
7.
Article in Chinese | WPRIM | ID: wpr-355275

ABSTRACT

<p><b>OBJECTIVE</b>To examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits.</p><p><b>METHODS</b>New Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas.</p><p><b>RESULTS</b>The level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas.</p><p><b>CONCLUSIONS</b>The transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.</p>


Subject(s)
Animals , Antibodies, Monoclonal , Antigens, Helminth , Metabolism , Fluorescent Antibody Technique , Gene Expression Profiling , Granuloma , Parasitology , Helminth Proteins , Metabolism , Liver , Parasitology , RNA, Messenger , Rabbits , Schistosoma japonicum , Schistosomiasis japonica
8.
Chinese Journal of Epidemiology ; (12): 181-185, 2015.
Article in Chinese | WPRIM | ID: wpr-335175

ABSTRACT

<p><b>OBJECTIVE</b>To study the dynamics of the reinfection of Schistosoma japonicum and related risk factors among the people in schistosomiasis endemic areas in China.</p><p><b>METHODS</b>Literature retrieval was conducted by using databases of PubMed, CNKI,VIP and Wanfang to collected all the data about the human re-infection of Schistosoma japonicum and related risk factors in the endemic areas in China. And a Mata-analysis was conducted on the literatures met the inclusion standards.</p><p><b>RESULTS</b>Eighteen studies involving 12 604 people for infection survey and 3 128 people for re-infection survey were included in the analysis. The overall infection rate was 20.8%, and the overall re-infection rate was 21.0% . The difference had no statistical significance (Z = 1.12, P = 0.26). The re-infection related factors included baseline infection intensity (OR = 3.58, 95% CI: 1.56-8.22); the index of contaminated water OR = 2.37, 95% CI: 1.08-5.22); distance from house to river-side (OR = 1.72, 95% CI: 0.41-7.30) and age (OR = 0.48, 95% CI: 0.19-1.23).</p><p><b>CONCLUSION</b>The baseline infection intensity, the index of contaminated water and distance from house to river-side were the risk factors related to the re-infection of Schistosoma japonicum and age was a protective factor.</p>


Subject(s)
Animals , Asians , China , Humans , Risk Factors , Schistosoma japonicum , Schistosomiasis japonica , Epidemiology , Parasitology
9.
Article in Chinese | WPRIM | ID: wpr-239212

ABSTRACT

<p><b>OBJECTIVE</b>To observe the dynamic changes of immune responses of splenocytes in mice immunized with recombinant vaccine Bifidobacterium bifidum (pGEX-Sj32) of Schistosoma japonicum and investigate the immunological mechanism of the vaccine.</p><p><b>METHODS</b>Eighty-eight BALB/c mice were randomized for immunization with 10⁶ CFU recombinant vaccine orally or with 10⁵ CFU recombinant vaccine intranasally. Four mice were selected from each group every two weeks to test the responses of the splenocytes to stimulations with SjAWA or ConA. MTT assay and flow cytometry were used to assess splenocyte proliferation and the distribution of CD4⁺ and CD8⁺ T cells, respectively; the levels of interleukin-10 (IL-10), IL-12 and tumor necrosis factor-α (TNF-α) in the cell culture supernatant were detected by ELISA.</p><p><b>RESULTS</b>Regardless of the stimulations, the splencytes showed significantly enhanced proliferation in weeks 2-16 in oral administration group and in weeks 2-18 in intranasal group (P<0.01). CD4⁺ subsets in both two groups increased obviously in weeks 2-12 (P<0.01) but CD8⁺ subsets remained stable. In oral administration group, the levels of TNF-α, IL-10 and IL-12 increased in weeks 2-14, 2-18 and 2-14, and peaked at week 8, 10 and 6, respectively; in intranasal group, the cytokines increased in weeks 2-14, 2-18 and 2-18, and peaked at week 8, 10 and 8, respectively.</p><p><b>CONCLUSION</b>The recombinant vaccine rBb (pGEX-Sj32) can induce effective immune responses in mice.</p>


Subject(s)
Animals , Antigens, Helminth , Allergy and Immunology , Bifidobacterium , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Interleukin-10 , Allergy and Immunology , Interleukin-12 , Allergy and Immunology , Mice , Mice, Inbred BALB C , Schistosoma japonicum , Schistosomiasis japonica , Spleen , Cell Biology , Allergy and Immunology , Tumor Necrosis Factor-alpha , Allergy and Immunology , Vaccination , Vaccines, Synthetic , Allergy and Immunology
10.
Chinese Journal of Biotechnology ; (12): 501-511, 2015.
Article in Chinese | WPRIM | ID: wpr-240624

ABSTRACT

To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.


Subject(s)
Animals , Antibodies, Helminth , Blood , Cloning, Molecular , DNA, Complementary , Escherichia coli , Female , Genes, Helminth , Helminth Proteins , Genetics , Allergy and Immunology , Immunoglobulin G , Blood , Male , Mice , Mice, Inbred BALB C , Real-Time Polymerase Chain Reaction , Recombinant Proteins , Allergy and Immunology , Schistosoma japonicum , Genetics , Schistosomiasis japonica , Vaccination
11.
Chinese Journal of Biotechnology ; (12): 1669-1678, 2014.
Article in Chinese | WPRIM | ID: wpr-345556

ABSTRACT

Radiation sensitive protein 23 (RAD23) is a nucleotide excision repair (NER) protein that plays an important role in Ubiquitin-proteasome pathway (UPP). Schistosoma japonicum radiation sensitive protein23 (SjRAD23) cDNA sequences were amplified by PCR and cloned into pET28a (+) vector to construct recombinant expression plasmid pET28a(+)-SjRAD23. The recombinant protein was expressed as both inclusion bodies and the supernatant in Escherichia coli BL21 (DE3) cell. Immunofluorescence observation shows that SjRAD23 was mainly distributed on the tegument surface of the worms. ELISA assay reveals that specific IgG, IgG1 and IgG2a antibodies could be detected in the sera of rSjRAD23 immunized mice. Western blotting analysis shows that the recombinant SjRAD23 could be recognized by serum specific to soluble adult worm antigen of S. japonicum. BALB/c mice vaccinated with rSjRAD23 combined with 206 adjuvant revealed 35.94% worm reduction and 40.59% liver egg reduction when compared with that of the adjuvant control


Subject(s)
Animals , Antibodies, Helminth , Blood , Blotting, Western , Cloning, Molecular , DNA Repair Enzymes , Genetics , Metabolism , DNA, Complementary , Enzyme-Linked Immunosorbent Assay , Escherichia coli , Genetic Vectors , Helminth Proteins , Genetics , Allergy and Immunology , Immunoglobulin G , Blood , Mice , Mice, Inbred BALB C , Recombinant Proteins , Genetics , Allergy and Immunology , Schistosoma japonicum , Genetics , Metabolism , Schistosomiasis japonica , Vaccines , Allergy and Immunology
12.
Mem. Inst. Oswaldo Cruz ; 108(7): 865-872, 1jan. 2013. tab, graf
Article in English | LILACS | ID: lil-696017

ABSTRACT

Schistosomiasis is an endemic parasite disease and praziquantel is the only drug currently in use to control this disease. Experimental and epidemiological evidence strongly suggests that Microtus fortis ( Mf ) is a naturally resistant vertebrate host of Schistosoma japonicum . In the present study, we found that Mf serum albumin ( Mf -albumin) and the conditioned medium of pcDNA3.1- Mf -albumin caused 46.2% and 38.7% schistosomula death rates in 96 h, respectively, which were significantly higher than that of the negative control (p < 0.05). We also found that mice injected with Mf -albumin had a 43.5% reduction in worm burden and a 48.1% reduction in liver eggs per gram (p < 0.05) in comparison to the control animals. To characterise the mechanisms involved in clearance, schistosomula were incubated with fluorescein isothiocyanate-labelled Mf -albumin and fluorescent enrichment effects were found in the gut lumen of schistosomula after 48 h of incubation. Next, digestive tract excretions from schistosomula were collected and the sensitivity of Mf -albumin to digestive tract excretions was evaluated. The results indicated that schistosomula digestive tract excretions showed indigestibility of Mf -albumin. The death of schistosomula could be partially attributed to the lack of digestion of Mf -albumin by digestive tract excretions during the development of the schistosomula stage. Therefore, these data indicate the potential of Mf -albumin as one of the major selective forces for schistosomiasis.


Subject(s)
Animals , Arvicolinae/parasitology , Schistosoma japonicum/drug effects , Serum Albumin/pharmacology , Chromatography, Affinity , Serum Albumin/isolation & purification
13.
Chinese Journal of Biotechnology ; (12): 814-822, 2013.
Article in Chinese | WPRIM | ID: wpr-233198

ABSTRACT

Schistosomiasis japonica is an endemic, zoonotic disease of major public health importance in China. Vaccination is needed as a complementary approach to the ongoing control programs. In the present study, we determined if the efficacies of DNA vaccine encoding the SjGST and Sj32 asparaginyl endopeptidase protein could be enhanced by boosting with SjGST-32 protein vaccines. Mice were inoculated with a VR1012-SjGST-32 DNA vaccine followed by boosting with rSjGST-32 at 0, 14 and 28 d. Two weeks after the final boost, mice were challenged percutaneously with cercariae. On day 45 following the challenge, all mice were sacrificed and the numbers of recovered worms and hepatic eggs were counted. Moreover, we analyzed the immune response among various vaccination groups. The results showed that DNA vaccine efficacy was enhanced when mice were boosted with protein vaccine. Adult worm and liver egg burdens were reduced 42.3% and 59.6%, respectively. We further found that DNA vaccine followed by boosting with protein significantly increased the IgG titer and T cell proliferation over those seen in mice vaccinated solely with DNA vaccines. Furthermore, the higher level of IFN-gamma expression in the splenetic CD4+ T cell showed that DNA prime-Protein boosting vaccine induced CD4+ Th1-type responses. Thus, DNA vaccine efficacy was significantly enhanced via boosting protein vaccine which might provide a basis for rational application of the Schistosoma vaccine.


Subject(s)
Animals , Antigens, Helminth , Allergy and Immunology , Female , Glutathione Transferase , Allergy and Immunology , Helminth Proteins , Allergy and Immunology , Immunization, Secondary , Methods , Mice , Mice, Inbred C57BL , Recombinant Fusion Proteins , Allergy and Immunology , Schistosoma japonicum , Schistosomiasis japonica , Vaccination , Methods , Vaccines, DNA , Allergy and Immunology
14.
Chinese Journal of Biotechnology ; (12): 891-903, 2013.
Article in Chinese | WPRIM | ID: wpr-233190

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.


Subject(s)
Animals , Antibodies, Helminth , Blood , Calcium-Binding Proteins , Genetics , Metabolism , Cloning, Molecular , Escherichia coli , Metabolism , Female , Genetic Vectors , Helminth Proteins , Genetics , Metabolism , Male , Mice , Recombinant Proteins , Genetics , Metabolism , Schistosoma japonicum , Genetics , Metabolism
15.
Article in English | WPRIM | ID: wpr-108246

ABSTRACT

Status and emerging issues in the use of praziquantel for treatment of human trematode and cestode infections are briefly reviewed. Since praziquantel was first introduced as a broadspectrum anthelmintic in 1975, innumerable articles describing its successful use in the treatment of the majority of human-infecting trematodes and cestodes have been published. The target trematode and cestode diseases include schistosomiasis, clonorchiasis and opisthorchiasis, paragonimiasis, heterophyidiasis, echinostomiasis, fasciolopsiasis, neodiplostomiasis, gymnophalloidiasis, taeniases, diphyllobothriasis, hymenolepiasis, and cysticercosis. However, Fasciola hepatica and Fasciola gigantica infections are refractory to praziquantel, for which triclabendazole, an alternative drug, is necessary. In addition, larval cestode infections, particularly hydatid disease and sparganosis, are not successfully treated by praziquantel. The precise mechanism of action of praziquantel is still poorly understood. There are also emerging problems with praziquantel treatment, which include the appearance of drug resistance in the treatment of Schistosoma mansoni and possibly Schistosoma japonicum, along with allergic or hypersensitivity reactions against praziquantel treatment. To cope with and overcome these problems, combined use of drugs, i.e., praziquantel and other newly introduced compounds such as triclabendazole, artemisinins, and tribendimidine, is being tried.


Subject(s)
Artemisinins , Benzimidazoles , Cestoda , Cestode Infections , Clonorchiasis , Cysticercosis , Diphyllobothriasis , Drug Resistance , Echinostomiasis , Fasciola , Fasciola hepatica , Humans , Hymenolepiasis , Hypersensitivity , Opisthorchiasis , Paragonimiasis , Phenylenediamines , Phosphatidylethanolamines , Praziquantel , Schistosoma japonicum , Schistosoma mansoni , Schistosomiasis , Sparganosis , Taenia , Taeniasis , Trematode Infections
16.
Article in Chinese | WPRIM | ID: wpr-267576

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of RNA interference (RNAi) on small heat shock protein (sHSP) Sjp40 of Schistosoma japonicum and its synergistic effect on the expression of SjHSP60, SjHSP70, and SjHSP90 mRNA, and observe the mRNA expression levels of Sjp40, SjHSP60, SjHSP70, and SjHSP90 in different stages of S.japonicum.</p><p><b>METHODS</b>Double-stranded RNA (dsRNA) of Sjp40 (dsSjp40) and a control dsRNA of green fluorescent protein (dsGFP) were generated by in vitro transcription and transfected into adult worm by immersing the worm in dsRNA solution. The total RNA and proteins were isolated simultaneously from the adult worms using TRIzol reagent 7 days after transfection. The expression levels of Sjp40, SjHSP60, SjHSP70, and SjHSP90 mRNA and the expression level of Sjp40 protein were determined by quantitative real-time PCR (qPCR) and Western blotting, respectively. The mRNA expression of HSPs of S. japonicum in different stages was evaluated by qPCR.</p><p><b>RESULTS</b>Compared with those in the control worms transfected with dsGFP, Sjp40 mRNA level was decreased by 80% in the worms transfected with dsSjp40, and the level of Sjp40 protein showed also a significant decrease. The mRNA expression levels of SjHSP60, SjHSP70, and SjHSP90 did not show an obvious synergism after Sjp40 RNAi. The expression profiles of Sjp40, SjHSP60, SjHSP70, and SjHSP90 showed significant differences in different stages of S. japonicum, and the expression level of Sjp40 mRNA in the egg stage was much higher than that of other HSP genes.</p><p><b>CONCLUSION</b>dsSjp40-RNAi can induce effective suppression of Sjp40 gene expression at both mRNA and protein levels, but no obvious synergism occurs in the mRNA expressions of SjHSP60, SjHSP70, and SjHSP90.</p>


Subject(s)
Animals , Gene Expression Profiling , Heat-Shock Proteins, Small , Genetics , Helminth Proteins , Genetics , RNA Interference , RNA, Double-Stranded , RNA, Messenger , Genetics , Schistosoma japonicum , Genetics
17.
Article in English | WPRIM | ID: wpr-819836

ABSTRACT

OBJECTIVE@#To assess the diagnostic efficacy of the currently most widely used indirect hemagglutination assay (IHA) and enzyme-linked immunosorbent assay (ELISA) for detection of Schistosoma japonicum human infections.@*METHODS@#A comprehensive search was undertaken from China National Knowledge Infrastructure, Wanfang Database, VIP Database, PubMed, Cochrane Library, Science Citation Index Expanded, Proquest, and the inclusion and exclusion criteria were strictly settled. The funnel plot was used to assess the publication bias, Cochran's Q test was employed to measure the homogeneity between studies, a summary receiver operating characteristic (SROC) curve was used to compare the diagnostic accuracy between the IHA and ELISA qualitatively by means of the Weighted Least Square method, the Ordinary Least Square method and the Robust regression method, and the diagnostic odds ratio (DOR) was drawn to compare the accuracy quantitatively.@*RESULTS@#Out of 785 publications, 19 papers were eventually selected for analysis. Literature quality assessment indicated that minor publication bias existed in studies pertaining IHA test, but no bias was found in literatures regarding ELISA test. The heterogeneity test showed a heterogeneity between studies was present (χ(2)=466.07 and 34.67, both P values<0.0001). The areas under the SROC curves of IHA were all higher than that of ELISA test using the three methods (Weighted Least Square method: 0.766 vs. 0.695, Ordinary Least Square method: 0.826 vs. 0.741, Robust regression: 0.815 vs. 0.715). The TPR* values for IHA and ELISA were 0.710, 0.759, 0.749, and 0.650, 0.686 and 0.666, respectively, and OR values were 5.997, 9.937, 8.893, and 3.432, 4.784 and 3.959, respectively. The DOR of IHA was 9.41 (95% CI: 4.88-18.18), and 4.78 (95% CI: 3.21-7.13) for ELISA.@*CONCLUSIONS@#All above results revealed that the diagnostic performance of IHA is better than that of ELISA. However, taking into account their unsatisfactory diagnostic value in areas with low infection intensity, a search for a better diagnostic test that can be applied in field situations in China should be given high priority.


Subject(s)
Animals , China , Epidemiology , Enzyme-Linked Immunosorbent Assay , Hemagglutination Tests , Methods , Humans , Least-Squares Analysis , Odds Ratio , ROC Curve , Schistosoma japonicum , Allergy and Immunology , Schistosomiasis japonica , Diagnosis , Epidemiology , Allergy and Immunology , Sensitivity and Specificity
18.
Article in English | WPRIM | ID: wpr-819818

ABSTRACT

OBJECTIVE@#To analyze the structure of aquaporins-3(AQP-3) from Schistosoma japonicum(SJAQP-3) using bioinformatical methods, and to provid of references for vaccine targets research.@*METHODS@#Protparam, BepiPred, TMHMM Server, MLRC, Geno3d, DNA star software packages were used to predict the physical and chemical properties, hydrophilicity plot, flexibility regions, antigenic index, surface probability plot, secondary structure, and tertiary structure of amino acid sequence of SJAQP-3.@*RESULTS@#SJAQP-3 had six transmembrane regions and two half-spanning helices that form a central channel. The half-spanning helices fold into the centre of the channel. Either of the half-spanning helix had a conserved motif of NPA common to all aquaporins. Predicted linear B-Cell epitopes were most likely at the N-terminal amino acid residues of 5aa-7aa, 59aa-62aa, 225aa-230aa, 282aa -288aa, 294aa -298aa and 305aa -307aa area. 59aa- 62aa, 225aa-230aa located outside the membrane, the others located inside the cell.@*CONCLUSIONS@#SJAQP-3 is a integral membrane protein in Schistosoma japonicum tegument. There are six potential epitopes in SJAQP-3. It might be a potential molecular target for the development of vaccines.


Subject(s)
Amino Acid Sequence , Animals , Aquaporin 3 , Allergy and Immunology , Computational Biology , Epitopes, B-Lymphocyte , Allergy and Immunology , Humans , Models, Molecular , Schistosoma japonicum , Genetics , Allergy and Immunology , Schistosomiasis japonica , Allergy and Immunology , Vaccination
19.
Mem. Inst. Oswaldo Cruz ; 106(7): 785-793, Nov. 2011. ilus
Article in English | LILACS | ID: lil-606640

ABSTRACT

Genome sequences for Schistosoma japonicum and Schistosoma mansoni are now available. The schistosome genome encodes ~13,000 protein encoding genes for which the function of only a minority is understood. There is a valuable role for transgenesis in functional genomic investigations of these new schistosome gene sequences. In gain-of-function approaches, transgenesis can lead to integration of transgenes into the schistosome genome which can facilitate insertional mutagenesis screens. By contrast, transgene driven, vector-based RNA interference (RNAi) offers powerful loss-of-function manipulations. Our laboratory has focused on development of tools to facilitate schistosome transgenesis. We have investigated the utility of retroviruses and transposons to transduce schistosomes. Vesicular stomatitis virus glycoprotein (VSVG) pseudotyped murine leukemia virus (MLV) can transduce developmental stages of S. mansoni including eggs. We have also observed that the piggyBac transposon is transpositionally active in schistosomes. Approaches with both VSVG-MLV and piggyBac have resulted in somatic transgenesis and have lead to integration of active reporter transgenes into schistosome chromosomes. These findings provided the first reports of integration of reporter transgenes into schistosome chromosomes. Experience with these systems is reviewed herewith, along with findings with transgene mediated RNAi and germ line transgenesis, in addition to pioneering and earlier reports of gene manipulation for schistosomes.


Subject(s)
Animals , Humans , Mice , Gene Transfer Techniques , Genome, Helminth/genetics , Schistosoma japonicum/genetics , Schistosoma mansoni/genetics , Animals, Genetically Modified , Chromosomes/genetics , Chromosomes/virology , DNA Transposable Elements , DNA, Helminth/genetics , DNA, Viral/genetics , DNA, Viral/isolation & purification , Genetic Vectors , Leukemia Virus, Murine/genetics , Leukemia Virus, Murine/isolation & purification , Membrane Glycoproteins/genetics , Membrane Glycoproteins/isolation & purification , RNA Interference , Schistosoma japonicum/virology , Schistosoma mansoni/virology , Viral Envelope Proteins/genetics , Viral Envelope Proteins/isolation & purification
20.
Mem. Inst. Oswaldo Cruz ; 106(7): 831-836, Nov. 2011. ilus, graf
Article in English | LILACS | ID: lil-606646

ABSTRACT

A real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is based on melting curve analysis of a hybrid between an amplicon from the S. japonicum internal transcribed spacer region 2 sequence, which is a 192-bp S. japonicum-specific sequence, and fluorophore-labelled specific probes. Real-time FRET PCR could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and a single egg inoculated in 100 mg of non-infected mouse faeces. All S. japonicum-infected snails and all faecal samples from infected mice were positive. Non-infected snails, non-infected mouse faeces and genomic DNA from other parasites were negative. This assay is rapid and has potential for epidemiological S. japonicum surveys in snails, intermediate hosts and faecal samples of final hosts.


Subject(s)
Animals , Mice , DNA, Helminth/analysis , Feces/parasitology , Schistosoma japonicum/genetics , Snails/parasitology , Fluorescence Resonance Energy Transfer , Nucleic Acid Hybridization , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Sensitivity and Specificity , Schistosoma japonicum/isolation & purification
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