Although great success has been achieved in schistosomiasis control,schistosomiasis japonica remains a public?health concern in China. Schistosoma japonicum is found to naturally infect over 40 mammalian animals. The implementation of the integrated schistosomiasis control strategy emphasizing infectious source control since 2004,which integrates replacement of bovines with machines,breeding domestic animals in fences,building safe pastures and chemotherapy of infected bovines,re?sults in a clear?cut reduction in the prevalence of S. japonicum infection in both humans and bovines,as well as the areas of in?fected snail habitats,and the national schistosomiasis control program is moving from transmission control towards transmission interruption and elimination. It has been found that goat is highly susceptible to S. japonicum infection,and previous epidemio?logical data have shown a high prevalence of infection in goat. However,the role of goat in the transmission of schistosomiasis ja?ponica has not been paid much attention,and there are few systematic surveys to evaluate the role of goat in schistosomiasis transmission in China to date. Professor Liang Yousheng’s group investigated S. japonicum development and reproduction(egg laying)in goat body,environmental contamination by goat feces,and the effect of temperature and humidity on the survival of S. japonicum eggs in goat feces. Their findings further demonstrate the role of goat in the transmission of schistosomiasis japonica. In addition,they proposed,based on their findings and previous reports,that the management of goat should be integrated into the national schistosomiasis control program in China,since goat is virtually one of the major infectious sources of schistosomia?sis japonica in China. Moreover,this group improved the fecal hatching test and optimized the parasitological technique for diag?nosis of S. japonicum infection in goats. These innovative studies fill in the gaps of goat schistosomiasis japonica research in Chi?na,and the research outcomes will enrich the currently implemented integrated schistosomiasis control strategy emphasizing in?fectious source control,and are believed to play a critical role in schistosomiasis elimination in China. Since schistosomiasis seri?ously affects goat husbandry development and local famer income,and goat has become a major infectious source of schistosomi?asis japonica in China,the control of goat schistosomiasis will facilitate the progress towards the elimination of schistosomiasis in China,and the following research priorities are suggested:(1)to emphasize the control of goat schistosomiasis,and to integrate the comprehensive management of goat into the national schistosomiasis control program in China;and(2)to develop new tech?niques,products and interventions for the control of goat schistosomiasis.

Objective:To explore the therapeutic effect of intra-articular injection of triptolide nanomaterials on rabbit antigen-induced knee arthritis.Methods:Twenty-seven New Zealand white rabbits were randomly divided into 4 groups. After antigen-induced arthritis (AIA) model were induced, the knee joints of triptolide nanomaterials (TPNA) group, triptolide (TP) group and betamethasone (BS) group were injected intra-articularly under ultrasound guidance with triptolide nanomaterials, triptolide and betamethasone respectively, 7 rabbits in each group. And the other 6 rabbits were punctured but not injected with any drugs as the control group. The pathological changes of joint swelling, synovitis and bone erosion were examined. Student's test, repeated measure data of analysis of variance (ANOVA), Mann-Whitney U test and Kruskal- Wallis test were used for statistical analysis. Results:① Before treatment, the knee joint diameters of the TPNA group, TP group, BS group and control group were (2.02±0.08) cm, (2.08±0.06) cm, (2.10±0.06) cm and (2.18±0.07) cm, respectively. After one week of administration, the knee joint diameters of the above groups were (1.85±0.06) cm, (1.89±0.07) cm, (1.93±0.08) cm and (2.15±0.08) cm, respectively. Knee joint swelling was significantly reduced in each treated group after a week of intra-articular injection. With the extension of treatment, the diameter of rabbit knee joints in each experimental group gradually decreased gradually ( F=58.83, P<0.01; F=53.78, P<0.01; F=68.24, P<0.01), and the diameter of rabbit knee joints in the TP group, TPNA group and BS group was significantly smaller than that of the control group ( F=63.83, P<0.01; F=71.94, P<0.01; F=140.79, P<0.01). ② The synovitis score of TP group was lower than that of the control group ( Z=-2.082, P<0.05), which was mainly mild synovitis. While the synovitis scores of TPNA group and BS group were lower than that of TP group ( Z=-2.082, P<0.05; Z=-2.687, P<0.05), most of which were free from synovitis. There was no statistical significant difference between BS group and TPNA group ( Z=-1.000, P>0.05). ③ The pathological scores of bone destruction in the TPNA group, TP group and BS group were all reduced compared with the control group ( Z=-2.505, P<0.05; Z=-2.216, P<0.05; Z=-2.505, P<0.05). There was no statistical significant difference between the TPNA group, TP group and BS group ( χ2=0.588, P>0.05). Conclusion:Intra-articular injection of triptolide nanomaterials can relieve joint swelling, reduce synovitis, and delay bone erosion. Its effect is similiar to glucocorticoid, better than simple triptolide. Triptolide nanomaterials have the potential to be an effective drug for arthritis by intra-articular injection.

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

The gene fragment encoding the egg-shell precursor protein of Schistosoma japonicum was amplified with RT-PCR by using PCR primer designed according to the 423 bp cDNA fragment of the Philippine strain of S.japonicum, the corresponding mDNA fragment of Chinese strain as template and then the 5' and 3' ends of this gene cDNA were amplified with 5' RACE and 3' RACE by using a series of primers designed according to the result of sequencing. Result of sequence analysis showed that this fragment, named as Sj423, contained a complete open reading frame (ORF) of gene encoding the egg-shell precursor protein of S.japonicum.(Chinese strain). As demonstrated by sequencing analysis. No intron could be detected in this gene fragment. This gene was subsequently expressed in E.coli after cloning into the expression vector pET28c(+). The molecular mass of the expressed product of this gene was 20.9 kDa as revealed by SDS-PAGE analysis, and Western blot analysis showed that the recombinant protein expressed could react well with the rabbit antiserum against the worm antigen of S.japonicum;indicating the good antigenicity of this expressed product.

Two recombinant plasmids pVAX/Sj23 and pVAX/mIL-18 containing Schistosoma japonicum 23 000 membrane protein (Sj23) and murine IL-18 were evaluated for their ability to induce immune responses and to protect against S. japonicum challenge in mice. All animals vaccinated with pVAX/Sj23 alone or plus pVAX/mIL-18 developed specific anti-SWAP (soluble worm antigen preparation) ELISA antibody and splenocyte proliferation response,and co-injection of pVAX/mIL-18 significantly increased the production of IFN-γ and IL-2 compared with pVAX/Sj23 alone, indicating that IL-18 enhances the Th1-dominant immune response. The challenge experiment showed that worm reduction rates in pVAX/Sj23 group compared with control group (pVAX1) was 26.5% and in the pVAX/Sj23 plus pVAX/mIL-18 group was 41.9% ,and the hepatic egg reduction rates were 42.7 and 49.6%,respectively. These results indicated that co-injection of an IL-18 plasmid with Sj23 DNA vaccine efficiently improves the protective effect against S. japonicum infection.

Objective To investigate the applicability of zebra fish thrombosis model in antithrombotic activity screening of Chinese materia medica.Methods The living zebra fish thrombosis model was induced by adrenaline hydrochloride. Zebra fish were randomly divided into blank control group, model group, positive medicine group and medication group. Each group was given the corresponding medicine or embryo culture water. O-anisidine staining solution was used to stain and calculate the staining intensity of erythrocytes in zebra fish heart, and quantitative analysis was carried out. The platelet aggregation of transgenic zebra fish was observed and under qualitative analysis. Results Compared with the model group, 100μg/mL salvianolic acid B, 300, 900μg/mL aqueous extract of Salvia miltiorrhiza, 45μg/mL 95% ethanol extract and 400, 1200μg/mL hypothalamus could significantly inhibited the formation of zebra fish thrombosis (P<0.01).ConclusionZebra fish thrombosis model has good applicability in antithrombotic activity screening of Chinese materia medica.

Objective To evaluate the effects of age factors on the pharmacokinetics of ropivacaine during combined lumbar plexus-sciatic nerve block in patients.Methods Twenty patients of both sexes,aged ≥ 18 yr,weighing 50-75 kg,of American Society of Anesthesiology physical status Ⅰ or Ⅱ,scheduled for elective lower extremity surgery,were divided into 2 groups (n=10 each) according to age:young and middle-aged group (18-59 yr) and elderly group (≥ 60 yr).Combined lumbar plexus-sciatic never block was performed using an ultrasonic instrument and a nerve stimulator.Lumbar plexus block was performed with 0.5％ ropivacaine 30 ml.Sciatic never block was performed with 2％ lidocaine 10 ml plus 0.75％ ropivacaine 10 ml.Before administration and at 5,10,15,20,30,45,60,120,180 and 360 min after administration,blood samples were collected from the radial artery for determination of the blood concentration of ropivacaine by high-performance liquid chromatography.The area under the concentrationtime curve,maximum concentration (Cmax),time to C terminal elimination half-life and clearance were calculated.Results Compared with young and middle-aged group,the blood concentration of ropivacaine was significantly decreased at 5-45 min after administration,the Cmax was significantly decreased,terminal elimination half-life was significantly prolonged (P＜0.05),and no significant change was found in area under the concentration-time curve,the time to C and clearance in elderly group (P＞ 0.05).Conclusion Age factors can affect the pharmacokinetics of ropivacaine during combined lumbar plexus-sciatic nerve block,and both absorption and metabolism of ropivacaine are slower in elderly patients than in young and middle-aged patients.

Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

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.
Animals ; Antibodies, Helminth ; blood ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Female ; Genes, Helminth ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin G ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; prevention & control ; Vaccination

Objective To clone,express and characterize a tegument protein gene of Schistosoma japonicum(Sj29),and investigate the immune protection of the recombinant protein against S.japonicum in mice.Methods The gene coding for Sj29 protein was amplified by PCR,and the sequence was analyzed by bioinformatics tools.Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced the recombinant with IPTG.The recombinant protein(rSj29) was purified by His-binding-resin affinity chromatography and characterized by Western blotting.Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval) respectively with 100 ?l recombinant rSj29(0.1 mg/ml),adjuvant or PBS.At the 15th day after the final inoculation,each mouse was challenged by 40 ?2 cercariae of S.japonicum.At the 53th day after infection,the mice were sacrificed to obtain the number of adult worms,number of eggs in liver and feces.Serum samples were collected at pre-immunization and certain time after immuniza-tion,and were analyzed for IgG by ELISA.The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique.mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR.Results A 576 bp Sj29 gene fragment was obtained.The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body.The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice.The number of adult worms(15.4?5.9),number of hepatic eggs(40 143.3?2 995.9) and number of fecal eggs(3 803.9?110.9) in re-combinant protein group were significantly higher than those of PBS control group(20?3.4,49 318.1?6 648.3,5 238.1? 303.5,respectively)(P