Objective To analyze the main B cell antigenic determinant of Cap protein of Porcine circovirus (PCV), construct a prokaryotic expression system for multi-antigenic determinant, and analyze the interest protein in order to provide a basis for the development of Porcine circovirus diagnostic reagent. Methods The PCV2 Cap gene was designed and synthesized, then connected to PET-28 a (+) vector and transformed into Escherichia coli BL21 (DE3) to construct a recombinant plasmid PET-28 (a) -Cap. The recombinant plasmid was induced to express the recombinant protein, which was then purified through His affinity chromatography. Results The recombinant plasmid was successfully expressed in Escherichia coli, whose weight was about17. 4 k Da, in agreement with the expectation. Western blot showed that the recombinant protein could specifically bind to Porcine circovirus positive sera. Conclusion The expression and purification of PCV2 Cap protein lays the foundation for the establishment of the diagnostic method of the PCV serum antibody, as well as effective monitoring of the immunity protection of PCV in farms.

Objective To construct the prokaryotic expression system for B cell epitopes of Toxoplasma gondii surface antigens (SAG)1 and 2,and analyze their immunoreactivity.Methods The B cell epitope sequences of Toxoplasma gondii SAG1 and SAG2 were designed and synthesized according to the gene sequences of SAG1 and SAG2 published in GenBank.The target gene was subcloned into pET-28a (+),and expressed in Escherichia coli BL21 (DE3).Complete single colonies were then selected from LB medium containing kanamycin.After culture under shaking to an absorbance (A450) value of 0.6-0.8,IPTG was added at a final concentration of 1 mmol/L to induce expression of the target gene.To obtain high concentrations of recombinant SAG (rSAG),the rSAG protein was purified using a His fusion protein purfication kit,and the resultant His-tagged purified protein was identified by Western blotting and ELISA.Results Results of PCR and double-enzyme digestion showed a specific protein band at ～909 bp.As expected,sequencing of the band further confirmed the construction of rSAG.SDS-PAGE revealed the rSAG molecular weight (Mr) of ～50 000.Western blotting showed specific binding of rSAG with a 1 ∶ 100 dilution of T.gondii-positive mouse serum.ELISA results showed a most positive reaction with the dilution of 1 ∶ 1 for recombinant protein and the dilution of 1:50 for positive serum.Moreover,reactions between different dilutions of rSAG and different concentrations of T.gondii-positive mouse serum,and a good performance in distinguishingnegative from positive serum.Conclusion The constructed recombinant protein rSAG is soluble and has good immunoreactivity.

In order to obtain recombinant E2 protein of Classical swine fever virus (CSFV), B cell antigen epitope was re-constructed and expressed in E. coli. The recombinant gene sequence was cloned into pET-28a(+) and inserted into BL21(DE3), The recombinant fusion protein was expressed with induction of IPTG and purified using Ni-NAT His?Bind Resin. Results showed that the reconstructed gene for multiple antigen epitope were about 500 bp, which was consistent with the results of PCR and double restriction digestion. Recombinant protein was soluble and molecular weight was about 23 kDa. Western blot showed that the recombinant protein reacted with positive swine fever antiserum. The recombinant protein may be used to develop diagnostic assays for swine fever.

The conceptual design of shared refrigerator is mainly from the humanization to solve the problem that different smells that made by different foods stored together, and it can effectively avoid the unreasonable use of the refrigerator space. In this refrigerator design, the practical problems encountered in the using process are fully considered and analyzed, and the humanized design concept is widely used. This article, focusing on people's needs in their daily lives, taking full account of the relationship between structure and function of the refrigerator, is mainly about the innovative design and research of shared refrigerator in terms of function, structure, materials, process and man-machine relationship, etc. This article puts forwards a new refrigerator–design-concept to make people feel the beauty, happiness and convenience of life brought by the design.

Through the analysis of current shoe cabinet market, aiming at some of the families that in needs of shoes cleaning, disinfection and storage, especially in areas of hot weather or frequently accompanied by rain and snow.The shoe cabinet was redesigned, to meet the needs of consumers on the shoes cleaning, sanitary and storage.This paper is mainly based on ergonomics principle and humanized design concept, designing and researching from the aspects of function, material, color and institutions of shoe cabinet, to design multi-functional shoe that sets disinfection and storage as a whole and give a new design concept for shoe cabinet which makes users to enjoy the multi-functional shoe cabinet with hygiene and soft at the same time feeling the beauty and enjoy what design brings to our life.

As science continues to progress, human civilization has ever enter into a new realm.Household electrification has become a symbol of progress .In southern China and some humid regions in foreign countries, clothes are still wet in several days after washing.People deeply suffered from it.The invention of clothing quick-drying device has solved this problem and has brought great convenience to people living in wet areas.According to the need of the people at home or traveling, this paper brought a new design conception which makes innovative design and research from the function and structure, materials and technology and human relationships.To let people use it at the same time feel a happy convenience life.

During development, Schistosoma japonicum undergoes many morphological and physiological transformations as a result of profound changes in gene expression. Proteins containing zinc finger motifs usually play an important role in DNA recognition, RNA packaging, and transcriptional activation. In our current study, we cloned the open reading frame (ORF) of SjZFP1 of S. japonicum, which encodes a zinc finger protein. We analyzed the complementary DNA (cDNA) sequence of SjZFP1 and examined the expression of SjZFP1 messenger RNA (mRNA) at various developmental stages. We also tested the effects of RNA interference (RNAi) silencing on worm burden, spawning, and egg hatching. The ORF in the SjZFP1 cDNA was 1017 bp in length and was predicted to encode a 338-aa protein with a molecular mass of approximately 38.5 kDa and theoretical isoelectric point (pI) of 7.08. Several conserved regions, including a B-box-type zinc-binding domain, two bipartite nuclear localization signal domains, a paired amphipathic helix repeat, and overlapping RING and PHD finger domains, were identified in the predicted amino acid sequence of SjZFP1. Using real-time PCR, we showed that the SjZFP1 mRNA was expressed across all of the developmental stages of the parasite and that the level of transcription was highest in the cercariae, eggs, schistosomula, and mature adult worms. The level of SjZFP1 mRNA expression in cultured schistosomula treated with one of two SjZFP1-specific small interfering RNAs (siRNAs; AY770 and AY546) was reduced by over 80 %, compared with that in the controls. In RNAi experiments in BALB/c mice, the level of SjZFP1 mRNA increased significantly when the mice were treated with the same SjZFP1-specific siRNAs during the early stages of infection. By contrast, the level of SjZFP1 mRNA decreased significantly when the mice were treated with the SjZFP1-specific siRNAs during the middle to late stages of infection. In four independent experiments, fewer worms were recovered from mice treated with the SjZFP1-specific siRNAs, compared with the number of worms recovered from the control mice. Both the average number and hatching rates of liver eggs recovered from mice treated with the SjZFP1-specific siRNAs during the middle to late stages of infection were significantly lower than those of the liver eggs recovered from the control mice. Our results suggest that the SjZFP1 gene might be important for parasite development, spawning in the vertebrate host, and egg hatching.

The innovative design of mobile cooking table is aimed to meet growing consumers’ demand, on the basis of existing technical conditions. Mobile table is a mobile cooking equipment. But this ‘mobility’ is not defined as the moving of table while working. In other words, it realizes the function of the mobile cooking according to the different operating environment. This paper mainly on the basis of the principle of ergonomics and the concept of humanized design, conduct the design on the overall size and construction of the cooking table from the aspect of function, material and institution to develop a cooking table with strong practicality in order to meet demands of different consumers.

Aiming at the problem of long cycle and high cost in designing power valve products at present, a nuclear power valve design method for collaborative design, simulation and optimization is put forward.Through the summarized characteristics of nuclear power valve products in CAE series, the basic framework of parametric CAE analysis is constructed.Combined with parametric variant design and simplification module for valve parts, the heterogeneous information between CAD and FEA of valve models is expressed by using the universal data exchange technology.According to the integrated development framework, an integrated system, which integrates parametric design, multi-characteristic simulation, and multidisciplinary design optimization for the nuclear power valves together supporting heterogeneous model transfer, is constructed and validated through a case.Through integration of heterogeneous CAD/CAE software such as UG, ANSYS and iSIGHT, the analytical efficiency is enhanced by coupling respective and unique advantage of various software systems.This transfer is benefit for mutual coordination and information sharing that improves the automation in design and analysis on the valve products.

This summary reported epidemiological investigations into domestic animal schistosomiasis that were performed in 2012 in China. During the whole year, Schistosma japonicum infection was examined in the following animals using miracidial hatching technique:684 899 cattle and buffalos, 71 473 goats and sheep and 142 976 other domestic animals from 7 provinces (Hunan, Hubei, Jiangxi, Anhui, Jiangsu, Sichuan and Yunnan). The positive animals included 3961 cattle and buffalos, 406 goats and sheep and 21 other domestic animals. The positive cattle and buffalos from Hunan and Jiangxi provinces accounted for 80.06%of total positive animals. These investigations showed a significant decrease in the prevalence of Schistosmajaponicum infection. However, Dongting Lake region in Hunan province and Poyang Lake region in Jiangxi province were still the key areas for control of domestic animal schistosomiasis.

Aiming at the problem of long cycle and high cost in designing power valve products at present, a nuclear power valve design method for collaborative design, simulation and optimization is put forward. According to the integrated development framework, the integrated platform is constructed, which integrates design, simulation, and optimization of nuclear power valves together supporting heterogeneous model transfer. Through integration of heterogeneous CAD/CAE software such as UG, ANSYS and iSIGHT, the analytical efficiency is enhanced by coupling respective and unique advantage of various software systems. This transfer is benefit for mutual coordination and information sharing that improves the automation in design and analysis on the valve products.

Schistosomiasis japonica is a major public health problem in China. Domestic animals play a major role in the transmission of Schistosoma japonicum to humans. To better understand the epidemiology of schistosomiasis japonica in domestic animals in the mountainous areas of China, we performed a 5-year longitudinal study of schistosomiasis in cattle and horses in Yunnan Province from 2009 to 2013. We also performed a concurrent drug-based intervention study in three settlement groups in Yunnan Province aimed at developing an effective means of controlling transmission in this region. The prevalence of infection in cattle fluctuated between 1.67% and 3.05% from 2009 to 2011, and monthly treatments of schistosome-positive animals reduced the prevalence to 0% (P < 0.05) from 2012 to 2013. Prior to the intervention, we found that schistosomiasis was prevalent from May to October, with the highest prevalence observed in June (10.00%). We surveyed for environmental schistosome contamination, and 94.29% of the miracidia found were from cattle. Our study showed that it is possible to eliminate schistosomiasis in domestic animals in the mountainous regions of China by monthly treating cattle and horses from schistosome-positive households from May to October.

Previous studies have shown that irradiated schistosome cercariae can induce higher immunoprotection. In the hosts, the radiation attenuated cercariae can neither mature nor cause serious pathological damage. Single-sex schistosoma perform similar behaviors but induce uninterrupted immune stimulation. Whether or not single-sex schistosoma can induce immune protection in hosts is unknown. To explore this problem, we first infected male BALB/c mice with single-sex male schistosoma and then examined its impact on the secondary schistosomiasis. BALB/c mice were randomly divided into two groups. Each mouse in test group was infected with 100 male cercariae and untreated mice used as controls. At 5 and 7 weeks post-infection, all mice in two groups were treated with praziquantel (300 mg/kg). Then at 9 weeks post-infection, all mice were infected with 40 sex-mixed cercariae. At 15 weeks post-infection, all mice were sacrificed and liver tissues and blood samples were collected for detection of eggs, pathological changes and seroconversion. The results showed that single-sex male worm caused significant reduction in eggs hatching rate and pathological damages in livers, indicating a certain immunoprotective effect in hosts.

Schistosomiasis is a worldwide parasitic disease, and while it can be successfully treated with chemotherapy, this does not prevent reinfection with the parasite. Adenovirus vectors have been widely used for vaccine delivery, and a vaccination approach has the potential to prevent infection with Schistosoma. Here, we developed a recombinant adenoviral vector that expresses Schistosoma japonicum inhibitor apoptosis protein (Ad-SjIAP) and assessed its immunoprotective functions against schistosomiasis in mice. Murine immune responses following vaccination were investigated using enzyme-linked immunosorbent assays (ELISA), lymphocyte proliferation, and cytokine assays. The protective immunity in mice was evaluated by challenging with S. japonicum cercariae. Our results indicated that immunization with the Ad-SjIAP in mice induced a strong serum IgG response against IAP including IgG1, IgG2a, and IgG2b. In addition, lymphocyte proliferation experiments showed that mice treated with Ad-SjIAP significantly increased the lymphocyte response upon stimulation with recombinant Schistosoma japonicum inhibitor apoptosis protein (rSjIAP). Moreover, cytokine assays indicated that vaccination of Ad-SjIAP significantly increased the production of interferon (IFN)-γ and IL-2 as compared to the corresponding control group. Furthermore, following the challenge with S. japonicum cercariae, the vaccine conferred moderate protection, with an average rate of 37.95% for worm reduction and 31.7% for egg reduction. Taken together, our preliminarily results suggested that schistosoma IAP may be a potential vaccine against S. japonicum and that adenoviral vectors may serve as an alternative delivery vehicle for schistosome vaccine development.

In 2012,a total of 8170 bovines,479 ovines and 569 other domestic animals were examined for schistsomiasis at eight national surveillance sites using a miracidial hatching technique.The prevalence was 0.92% in bovines,1.88% in ovines and 0% in other animals.Among 89 infected bovines,54 bovines(72.00%) were younger than 3 years and 61 bovines(81.30%) showed low-intensity infection.The field feces and snails collected at the sites in Hunan and Jiangxi provinces were found positive for Schistosoma japonicum.The results suggested that the control of bovine schistsomiasis in the Dongting Lake region in Hunan Province and the Poyang Lake region in Jiangxi Province was a key and difficult point for the national agricultural schistosomiasis program.

In order to meet the goal of domestic animal schistosomiasis control by 2015 as proposed by the State Council of China,a survey of combining questionnaires and site visits was carried out in 28 counties in Hunan,Hubei,Jiangxi,Anhui,Sichuan and Yunnan provinces for evaluating the current situations and major risk factors.The survey realized serious risks if no special measures would be implemented.Based on the statistical analysis of 20 risk factors,13 control measures and degrees of controllability,five recommendations were proposed for risk control of domestic animal schistosomiasis.

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 Ca2 + 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.

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.

A full-length cDNA encoding the Schistosoma japonicum translocase of outer mitochondrial membrane 34(SjTOM34) was first isolated from 42-day-schistosome cDNAs.The cDNA had an open reading frame(ORF) of 1 083 bp and encoded 360 amino acids.Real-time quantitative RT-PCR analysis revealed that the expression of SjTOM34 was a little higher in 35-day schistosomes than worms in other stage.The SjTOM34 was subcloned into pET28(+) and expressed as both inclusion bodies and supernatant in Escherichia coli BL21(DE3) cells.Western blotting showed that the recombinant SjTOM34(rSjTOM34) had good immunogenicity.Immunolocalization analysis found that SjTOM34 was distributed all over the worms of S.japonicum.It's suggested that SjTOM34 may be important in schistosome,and further investigations are required to fully understand the function of this molecule.

In a previous study,the immunoprotective effect of UDP-Glucose-4-Epimerase protein of Schistosoma japonicum(SjGALE) was demonstrated,especially in reducing female spawning.In the present study,a combined use of rSjGALE and montanide ISA 206 was tried to see more potent immunoprotection.The results showed that the combination of rSjGALE and montanide ISA 206 induced specific antibodies and reduced liver egg burden,female spawning and hatching rate.However,their immunoprotective activity was not as stable as Freund adjuvant.