Molluscicidal effect of green synthesized silver nanoparticles using Azadirachta indica on Biomphalaria alexandrina snails and Schistosoma mansoni cercariae

Objective:To assess the molluscicidal effect of the eco-friendly green synthesized neem silver nanoparticles(neem-Ag NPs)against Biomphalaria alexandrina,the snail intermediate host for Schistosoma mansoni,and their cercaricidal potential.Methods:Methanol extracts from neem fruits were used for green synthesis of neem-Ag NPs.The neem-Ag NPs were characterized using UV-visible absorption spectra,dynamic laser light scattering technique,and transmission electron microscopy.The potential molluscicidal effect against adult and juvenile Biomphalaria alexandrina and the effect of the sub-lethal concentration on hatching of snail eggs and Schistosoma mansoni cercariae were evaluated.Results:The surface plasmon resonance of neem-Ag NPs showed a sharp absorption peak atλ_(max)=518 nm together with multiple peaks.The hydrodynamic diameter was(77.15±34.53)nm,the polydispersity index(0.338±0.000)and the zeta-potential-14.07 mV.Moreover,transmission electron microscopy showed that the average size of the nanoparticles was(27±2)nm.Agglomeration was evident and a light-colored capping layer could be seen coating the nanoparticles.Juvenile snails(LC_(50):0.83 ppm)were more susceptible to neem-Ag NPs than adults(LC_(50):1.07 ppm).In addition,neem-Ag NPs and neem at LC_(50)concentrations inhibited the egg-hatching of snails and showed cercaricidal activity in a time-dependent manner.Conclusions:Neem-Ag NPs have lethal activities against Biomphalaria alexandrina snails and their eggs,as well as Schistosoma mansoni cercariae.Hence,neem-Ag NPs could be a potential agent to control schistosomiasis.

Prevalence and Clinical Relevance of Schistosoma mansoni Co-Infection with Mycobacterium tuberculosis: A Systematic Literature Review

Tuberculosis disease stands for the second leading cause of death worldwide after COVID-19, most active tuberculosis cases result from the reactivation of latent TB infection through impairment of immune response. Several factors are known to sustain that process.Schistosoma mansoni, a parasite of the helminth genus that possesses switching power from an immune profile type Th1 to Th2 that favors reactivation of latent TB bacteria. The aim of the study was to assess the prevalence of the co-infection between the two endemic infections. Systematic literature was contacted at the University Clinical Research Center at the University of Sciences, Techniques, and Technologies of Bamako in Mali. Original articles were included, and full texts were reviewed to assess the prevalence and better understand the immunological changes that occur during the co-infection. In total, 3530 original articles were retrieved through database search, 53 were included in the qualitative analysis, and data from 10 were included in the meta-analysis. Prevalence of the co-infection ranged from 4% to 34% in the literature. Most of the articles reported that immunity against infection with helminth parasite and more specifically Schistosoma mansoni infection enhances latent TB reactivation through Th1/Th2. In sum, the impact of Schistosoma mansoni co-infection with Mycobacterium tuberculosis is under-investigated. Understanding the role of this endemic tropical parasite as a contributing factor to TB epidemiology and burden could help integrate its elimination as one of the strategies to achieve the END-TB objectives by the year 2035.

Bladder stones in a closed diverticulum caused by Schistosoma mansoni: A case report

BACKGROUND Genitourinary(GU)schistosomiasis is a chronic infection caused by a parasitic trematode,with Schistosoma haematobium(S.haematobium)being the prevalent species.The disease has a variable prevalence around the world,with a greater burden on,but not limited to Africa,South America,Asia,and the Middle East.CASE SUMMARY We report the case of a 30-year-old man who presented with symptoms of bladder stones.During endoscopic cystolithalopaxy,we did not detect any stones in the bladder.Upon careful scanning of the urinary bladder trigone,sandy patches were detected.We performed endoscopic resection,which revealed a closed diverticulum with bladder stones.The diverticular wall was sent for histopathology and revealed features of chronic granulomatous inflammation with numerous embedded Schistosoma eggs,with some of the eggs having lateral spines.The patient was treated with praziquantel,and his symptoms completely resolved.CONCLUSION GU schistosomiasis is primarily caused by S.haematobium.However,Schistosoma mansoni mediated GU schistosomiasis is unusual,making this a quite interesting case.

Anti-schistosomal activities of Echinops kebericho Mesfin root and Hagenia abyssinica (Bruce) J.F Gmel flower part crude extracts in Swiss albino mice

Objective: To evaluate the in vivo antischistosomal activities of the crude extracts of Echinops kebericho Mesfin(E. kebericho) root and Hagenia abyssinica(Bruce) J.F. Gmel(H. abyssinica) flower. Methods: Mice were infected with(150 ± 10) Schistosoma mansoni cercariae by paddling technique. Crude extracts were administered orally for five consecutive days at doses of 300, 600 and 1 200 mg/kg/day along with 200 mg/kg/day praziquantel and 3% tween 80 given as a control. Results: E. kebericho root extract showed a statistically significant(P < 0.05) reduction in fecal egg count of 64.44%, 42.96% & 26.82% and worm burden of 65.71%, 47.86% & 31.43% at treatment doses of 1 200 mg/kg/day, 600 mg/kg/day and 300 mg/kg/day, respectively. Similarly, H. abyssinica flower extracts showed a significant(P < 0.05) reduction in fecal egg count up to 84.57%, 77.06% & 63.89% and worm burden of 91.43%, 81.43% & 70.71% at a respective dose levels. In addition, a significant(P < 0.05) reduction in liver granuloma score was observed in all H. abyssinica administered dose groups and E. kebericho at 1 200 mg/kg/day dose group as compared to infected untreated control group. Conclusions: H. abyssinica and E. kebericho crude extracts show a promising antischistosomal activity.

Exogenous bone morphogenetic protein-7 reduces hepatic fibrosis inSchistosoma japonicum-infected micevia transforming growth factor-β/Smad signaling

AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided into three groups, including a control group (group A, n = 20), model group (group B, n = 20) and BMP-7 treated group (group C, n = 20). The mice in group B and group C were abdominally infected with S. japonicum cercariae to induce a schistosomal hepatic fibrosis model. The mice in group C were administered human recombinant BMP-7. Liver samples were extracted from mice sacrificed at 9 and 15 wk after modeling. Hepatic histopathological changes were assessed using Masson's staining. Transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), phosphorylated Smad2/3 (pSmad2/3) and Smad7 protein levels and localization were measured by Western blotting and immunohistochemistry, respectively, and their mRNA expressions were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). RESULTS: The schistosomal hepatic fibrosis mouse model was successfully established, as the livers of mice in group B and group C showed varying degrees of typical schistosomal hepatopathologic changes such as egg granuloma and collagen deposition. The degree of collagen deposition in group C was higher than that in group A (week 9: 22.95±6.66vs 2.02±0.76; week 15: 12.84±4.36 vs 1.74±0.80; P<0.05), but significantly lower than that in group B (week 9: 22.95±6.66 vs 34.43±6.96; week 15: 12.84±4.36 vs 18.90±5.07;P<0.05) at both time points. According to immunohistochemistry data, the expressions of α-SMA, TGF-β1 and pSmad2/3 protein in group C were higher than those in group A (α-SMA: week 9: 21.24±5.73 vs 0.33±0.20; week 15: 12.42±4.88 vs 0.34±0.27; TGF-β1: week 9: 37.00±13.74 vs 3.73±2.14; week 15: 16.71±9.80 vs 3.08±2.35; pSmad2/3: week 9: 12.92±4.81 vs 0.83±0.48; week 15: 7.87±4.09 vs 0.90±0.45; P<0.05), but significantly lower than those in group B (α-SMA: week 9: 21.24±5.73 vs 34.39±5.74; week 15: 12.42±4.88 vs 25.90±7.01; TGF-β1: week 9: 37.00±13.74 vs 55.66±14.88; week 15: 16.71±9.80 vs 37.10±12.51; pSmad2/3: week 9: 12.92±4.81 vs 19.41±6.87; week 15: 7.87±4.09vs 13.00±4.98;P<0.05) at both time points; the expression of Smad7 protein in group B was higher than that in group A and group C at week 9 (8.46±3.95 vs 1.00±0.40 and 8.46±3.95 vs 0.77±0.42; P<0.05), while there were no differences in Smad7 expression between the three groups at week 15 (1.09±0.38 vs 0.97±0.42 vs 0.89±0.39; P>0.05). Although minor discrepancies were observed, the results of RT-PCR and Western blotting were mainly consistentwith the immunohistochemical results. CONCLUSION: Exogenous BMP-7 significantly decreased the degree of hepatic fibrosis in both the acute and chronic stages of hepato-schistosomiasis, and the regulatory mechanism may involve the TGF-β/Smad signaling pathway.

Bioinformatics analysis of the structure and linear B-cell epitopes of aquaporin-3 from Schistosoma japonicum

Objective:To analyze the structure of aquaporins-3(AQP-3) from Schistosoma japonicum(SJAQP-3) using bioinformalical 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 Saa-7aa,59aa- 62aa,225aa-230aa,282aa -288aa,294aa -29Saa and 305aa -307aa area.59aa- 62aa,22Saa-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 SJ AQP-3.It might be a potential molecular target for the development of vaccines.

Schistosoma mansoni proteins attenuate gastrointestinal motility disturbances during experimental colitis in mice

AIM:To investigate the therapeutic effect of Schistosoma mansoni(S.mansoni) soluble worm proteins on gastrointestinal motility disturbances during experimental colitis in mice. METHODS:Colitis was induced by intrarectal injection of trinitrobenzene sulphate(TNBS) and 6 h later,mice were treated ip with S.mansoni proteins.Experiments were performed 5 d after TNBS injection.Inflammationwas quantified using validated inflammation parameters. Gastric emptying and geometric center were measured to assess in vivo gastrointestinal motility.Peristaltic activity of distal colonic segments was studied in vitro using a modified Trendelenburg set-up.Cytokine profiles of T-lymphocytes isolated from the colon were determined by real time reverse transcriptase-polymerase chain reaction. RESULTS:Intracolonic injection of TNBS caused severe colitis.Treatment with S.mansoni proteins significantly ameliorated colonic inflammation after 5 d.TNBS did not affect gastric emptying but significantly decreased the geometric center and impaired colonic peristaltic activity 5 d after the induction of colitis.Treatment with S.mansoni proteins ameliorated these in vivo and in vitro motility disturbances.In addition,TNBS injection caused a downregulation of effector T cell cytokines after 5 d,whereas a S.mansoni protein effect was no longer observed at this time point. CONCLUSION:Treatment with S.mansoni proteins attenuated intestinal inflammation and ameliorated motility disturbances during murine experimental colitis.

Modeling and analysis of Schistosoma Argonaute protein molecular spatial conformation

Objective:To analyze the amino acid sequence composition,secondary structure,the spatial conformation of its domain and other characteristics of Argonaute protein.Methods:Bioinformatics tools and the internet server were used.Firstly,the amino acid sequence composition features of the Argonaute protein were analyzed,and the phylogenetic tree was constructed.Secondly,Argonaute protein's distribution of secondary structure and its physicochemical properties were predicted.Lastly,the protein functional expression form of the domain group was established through the Phyre-based analysis on the spatial conformation of Argonaute protein domains.Results:593 amino acids were encoded by Argonaute protein,the phylogenetic tree was constructed,and Argonaute protein's distribution of secondary structure and its physicochemical properties were obtained through analysis.In addition,the functional expression form which comprised the N-terminal PAZ domain and C-terminal Piwi domain for the Argonaute protein was obtained with Phyre.Conclusions:The information relationship between the structure and function of the Argonaute protein can be initially established with bioinformatics tools and the internet server,and this provides the theoretical basis for further clarifying the function of Schistosoma Argonaute protein.

Liver cirrhosis and splenomegaly associated with Schistosoma mansoni in a Sudanese woman in Malaysia:A case report

We report a case of a patient with Schistosoma mansoni infection who presented with liver cirrhosis and splenomegaly.She was diagnosed by a serological test and Kato-Katz thick smear stool examination.The patient was a 52-year-old woman from Sudan who came to Malaysia for a week to visit her sons.The patient lives in the middle of Rabak region,Sudan,a highly endemic area for schistosomiasis where her daily routine includes rearing of cows and farming.The site of toilet and sources of drinking water are canals and wells;both infested with snails.Patient had a long history of exposure and coming into contact with water from these canals and wells.

Schistosoma japonicum attenuates dextran sodium sulfateinduced colitis in mice via reduction of endoplasmic reticulum stress

AIM To elucidate the impact of Schistosoma(S.) japonicum infection on inflammatory bowel disease by studying the effects of exposure to S. japonicum cercariae on dextran sodium sulfate(DSS)-induced colitis. METHODS Infection was percutaneously established with 20 ± 2 cercariae of S. japonicum, and colitis was induced by administration of 3% DSS at 4 wk post infection. Weight change, colon length, histological score(HS) and disease activity index(DAI) were evaluated. Inflammatory cytokines, such as IL-2, IL-10 and IFN-γ, were tested by a cytometric bead array and real-time quantitative polymerase chain reaction(RT-PCR). Protein and m RNA levels of IRE1α, IRE1β, GRP78, CHOP, P65, P-P65, P-IκBα and IκBα in colon tissues were examined by Western blot and RT-PCR, respectively. Terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling positive cells, cleaved-caspase 3 expression and Bcl2/Bax were investigated to assess the apoptosis in colon tissues.RESULTS Mice infected with S. japonicum cercariae were less susceptible to DSS. Mice infected with S. japonicum cercariae and treated with DSS showed decreased weight loss, longer colon, and lower HS and DAI compared with mice treated with DSS alone. A substantial decrease in Th1/Th2/Th17 response was observed after infection with S. japonicum. Endoplasmic reticulum(ER) stress and the nuclear factor-kappa B(NF-κB) pathway were reduced in mice infected with S. japonicum cercariae and treated with DSS, along with ameliorated celluar apoptosis, in contrast to mice treated with DSS alone. CONCLUSION Exposure to S. japonicum attenuated inflammatory response in a DSS-induced colitis model. In addition to the Th1/Th2/Th17 pathway and NF-κB pathway, ER stress was shown to be involved in mitigating inflammation and decreasing apoptosis. Thus, ER stress is a new aspect in elucidating the relationship between helminth infection and inflammatory bowel disease(IBD), which may offer new therapeutic methods for IBD.

Schistosoma haematobium and Plasmodium falciparum coinfection with protection against Plasmodium falciparum malaria in Nigerian children

Objective:Malaria remains the single leading killer of children in sub - Sahara Africa and Schistosomiasis is considered to be second to malaria in global importance.Co - infection of malaria and urinary schistosomiasis has been reported to exacerbate disease morbidity such as anaemia.In different part of the globe,the co - infection between malaria and schistosomiasis provides some protections on the infected persons.The protective effect of this co - infection elucidated immunologically using cytokines is lacking in our locality.Methods:Urine and blood samples obtained from the 160 volunteers were subjected to standard parasitological techniques for diagnosis of urinary schistosomiasis and malaria respectively.Blood samples collected from these volunteers comprising 80 children with schistosomiasis and malaria and the 80 children who had malaria only were subjected to cytokines concentration determination using commercial standard enzyme linked immunosorbent assay kits(Abeam,UK).Results:Eighty participants with co - infection had a mean malarial parasitaemia of 662±201.1μL while the 80 participants with only P.falciparum malaria had a mean malarial parasiteamia of 5943±3270.7μL.Also the volunteers had mean haemoglobin of 11.2 g/dL for co - infected individuals and 5.7 g/dL for participants with single infection of malaria.The serum cytokine levels of the children with S. haematobium and P.falciparum and only P.falciparum infection are as follows;interleukin - 4(16.6 pg/ mL versus 5.2 pg/mL),IL - 5(501.3 pg/mL versus 357.5 pg/mL);IL -8(2 550 pg/mL versus 309 pg/mL),IL - 10(273 pg/mL versus 290 pg/mL),TNF -α(25 pg/mL versus 290 pg/mL) and IFN -γ(21.9 pg/mL versus 2.5 pg/mL).The TNF -α/IL - 10 ratio is 7 for the children with co - infection while those with only P.falciparum malaria infection had a TNF -α/IL - 10 ratio of 0.9.Conclusion:We conclude that the elevated IL - 4,IL - 5,IL - 8 and IFN -γconcentration induced by schistosomiasis altered the Th1/Th 2 profile and protected the children against the morbidity and severity of malaria attack among the children with co - infection.

Orthotopic liver transplantation from a donor with Schistosoma japonicum

To the Editor:Despite of the rapid increase of donation after cardiac death(DCD)in China,the shortage of organs continues to be a major problem.Every organ procured is so valu able that it should never be discarded easily,especially a liver that could save a patient’s life in an emergency.This leads to the use of grafts from donors with unrecognized and unusual diseases,including schistosomiasis.[1]Here we reported a case of orthotopic liver

Immunomodulatory effect of garlic oil extract on Schistosoma mansoni infected mice

Objective:To assess the effect potency,and the immunomodulatory response of garlic oil extract in enhancing the host's immune system against the disorders caused by Schistosoma mamoni(S.mamoni) in mice at different stages of worm maturation.Methods:A total of 70 male CD-I Swiss albino mice were divided into 7 groups.Group Ⅰ:healthy control.Group Ⅱ:garlic oil group orally administrating 100 mg garlic oil extract /kg b.wt.3 d a week for 6 weeks.Group Ⅲ:infected with S.inansoni cercariae and left untreated for 42 d.Group Ⅳ:treated with garlic oil extract from day 1 to day 7 post infection(PI).Group Ⅴ:treated with garlic oil extract from day 14 till day 21 PI.Group Ⅵ:administrating garlic oil extract from day 35 until day 42 PI.Group Ⅶ received oil extract from the first day of infection for 42 d.Results:Garlic oil extract showed changes in the parasite tegument with a significant decrease in worm burden,hepatic and intestinal ova count with a decline in granuloma number and diameter.These alterations were accompanied with a reduction in serum TNF- α,ICAM-1,IgG and IgM after 7 and 42 d post S.mamoni cercarial infection.Conclusions:Results obtained confirmed the effect of garlic oil extract on the larval and mature stage of the parasite and in enhancing the host's immune system against the disorders caused by 5.mansoni in mice.

Schistosoma mansoni infection prevalence and associated risk factors among schoolchildren in Demba Girara,Damot Woide District of Wolaita Zone,Southern Ethiopia

Objective:To establish the prevalence and associated risk factors of Schistosoma mansoni(S.mansoni) infection among schoolchildren at a village in Wolaita Zone.Sothern Ethiopia,Methods:A cross-sectional study was carried out among primary schoolchildren.A total of 384 randomly selected study subjects provided stool samples for parasitological examination by Kato-Katz and Formalin-Ether concentration techniques.Secondary parasitological data were obtained from Health Center Laboratory to see the previous history of.S.mansoni infection in the area.Statistical analysis was performed using SPSS software version 16.Results:From the total children examined.85.4% were found positive for at least one helminth infection.S.mansoni infection(81.3% ) was the most prevalent and the prevalence of STH was 32% ..Moderate and heavy infection intensities were only observed in S,mansoni infections.The overall heavy intensity of infection was 56.4% .Contact to Bisarc stream was the most important factor for S.mansoni infection(OR 3.9) followed by herding cattle near the stream(OR2.527).Males were twice more likely to get the infection than females(OR 1.923).Analysis of secondary parasitological data showed that S.mansoni infection was a leading helminthic infection over the past years.Conclusions:The present study found a higher intensity and prevalence of S.mansoni infection in a rural village of Wolaita Zone.Therefore,appropriate integrated control and prevention measures need to be implemented in the study area.

Aristolochia gehrtii inhibits liver toxicity and apoptosis in Schistosoma malayensis infection

Objective:To evaluate a protective effect of Aristolochia gehrtii(A.gehrtii)leaves to inhibit liver toxicity and apoptosis in Schistosoma malayensis(S.malayensis)infection.Methods:Forty male albino mice were divided into four equal groups:group 1 control including noninfected healthy mice and groups 2,3&4 subcutaneously infected with S.malayensvs cercariae where groups 3&4 pretreated with A.gehrtii leaves(200 mg/kg,bwt)&cinnamoylamide(250mg/kg,bwt),respectively.Results:5.malayensis caused a significant increase in serum AST,ALT,ALP,MDA,NO,bilirubin,urea,creatinine,total cholesterol,LDL,triglycerides,and HDL levels.The pretreatment of A,gehrtii leaves and cinnamoylamide significantly inhibited that increase.On the other hand,S.malayensis induced a significant decrease in serum total protein,albumin,globulin,albumin/globulin ratio,blood SOD and GPx,while A.gehrtii leaves and cinnamoylamide pretreatment increased the above parameters.Treatment with A.gehrtii leaves and cinnamoylamide to S.malayensis infected mice increased p53 expression but decreased bcl-2expression.These results were supported by hislopalholqgical investigations.Conclusions:A.gehrtii inhibits liver toxicity and apoptosis in S.malayensvs infection and this effect is associated with the major cinnamoylamide ingredient of A.gehrtii leaves.

Perforation of small bowel caused by Schistosoma japonicum : A case report

A 67-year-old man from Jingzhou was admitted to the First Hospital Affiliated to Yangtze University in July 2013 with sudden onset of abdominal pain with dizziness for 12 h.The patient had sign of peritoneal irritation.Ultrasonography of the abdomen and pelvis showed hepatic fibrosis due to schistosomiasis.Computed tomography showed free gas in the peritoneal cavity.Plain abdominal radiography showed bilateral subdiaphragmatic accumulation of gas, perforation of the viscus, and radio-opacity in the left renal area.The patient underwent emergency exploratory laparotomy.At laparotomy, a moderate amount of muddy yellow pus was found in the intra-abdominal cavity.At the junction of the jejunum and ileum, about 250 cm from Treitz's ligament, there was an about 10-cm length of inflamed small bowel with perforation(3 mm in diameter) along the mesenteric border at the middle of the lesion.The patient underwent resection of the affected intestinal segment, along with end-to-end intestinal anastomosis.Histopathological examination revealed mucosal necrosis and hemorrhage with a large number of infiltrating eosinophils and neutrophils, and acute submucosal inflammation with a large number of infiltrating eosinophils and neutrophils associated with Schistosoma japonicum(S.japonicum) eggs.No intravascular adult parasite was found.Postoperatively, the patient was treated with praziquantel(30 mg/kg daily) for 4 d.The patient progressed well.To the best of our knowledge, this is the first case of small bowel perforation associated with eggs of S.japonicum.

Effect of artmether,hemin and Fe^(3+) on recombinant lactate dehydrogenase from Schistosoma japonicum

Objective:To explore antischitosome effects of artemether,hemin and Fe on S/LDH.Methods: Enzyme activity of rS/LDH was assayed in the standard reaction system by adding different concentration of reagents(0.00-0.10 mM artemether,0.00-0.02 mM hemin,0.00-0.50 mM Fe3+). Same solvents of the each reagent were used as control.Results:There was no enzyme activity inhibition observed at 0.10 mM artemther:obivious inhibition for lactate oxidation reaction and pyruvate reduction reaction were detected at 0.002 mM and 0.004 mM of hemin,respectively: comparing with that of the control（P【0.05）.The relative enzymatic activity inhibitions for pyruvate reduction reaction and lactate oxidation reaction at 0.02 mM hemin were 93.48%and 100.00%,respectively,comparing with that of the control（P【0.01）:both pyruvate reduction and lactate oxidation reaction were inhibited completely at 0.50 mM Fe3+,comparing with that of the control（P【0.01）.Conclusions:The results implied that SjLDH was not the direct molecular target of artemether.Hemin and Fe are inhibitors of SjLDH.

Studies on DNA Vaccine of the Conservative Region of Gene Encoding the Male-Specific Gynecophoral Canal Protein of Schistosoma japonicum (Chinese strain)

In order to observe the protective effect induced by vaccinating animals with the DNA vaccine of Sex-specific expression gene of Schistosoma, A 868 bp cDNA fragment amplified by RT-PCR from adult Schistosoma japonicum (Chinese strain) mRNA was cloned into the eukaryotic expression vector pcDNA3 and the recombinant eukaryotic expression vector pcDNA3-SjGCP1 was directly injected into BALB/c mice intramuscularly 3 times with the interval of 3 weeks .Both the vaccinated and control group of mice were challenged with 40 cercariae of Sj 5 weeks after last injection and perfused 7 weeks post-challenge. The worm and egg reduction rate got from vaccinated mice was 32.4% and 46.9% respectively. The result indicated that pcDNA3-SjGCP1 DNA vaccine induces the significant protection in animal against Schistosoma japonicum infection.

The Construction of Schistosoma Japonicum Vaccine BCG-Sj26GST and Its Identification

Summary: The expression of foreign gene, Schistosoma Japonicum 26 ku antigen (Sj26GST), in Bacillus Calmette Guerin (BCG), Mycobacterium ( M. smegmatis ) and Escherichia coli ( E. coli ) were studied. The cDNA fragment encoding Sj26GST was amplified by PCR using plasmid pGEX, which could express Sj26GST in E. coli as template. The Sj26GST cDNA was cloned into the downstream of human M. tuberculosis heat shock protein (hsp) 70 promoter with correct reading frame, and then the DNA fragment containing hsp70 promoter and Sj26GST gene were subcloned together into E. coli Mycobacteria shuttle plasmid pBCG 2000 to construct the expression shuttle plasmid pBCG Sj26. The recombinant BCG and M. smegmatis mc 2 155, which were electroplated with pBCG Sj26, could express Sj26GST and the recombinant Schistosoma Japonicum vaccine BCG Sj26GST was made. The recombinant Sj26GST (rSj26GST) were soluble and could be observed on SDS PAGE at molecular weight of 26 ku. The content of rSj26GST accounted for 15 % and 10 % of total bacterial protein in BCG and M. smegmatis respectively. The results of Western blot showed the combination of rSj26GST with antibody of GST.

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

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