Tetraspanin 2-A(SjTsp2-A) gene was amplified by PCR.pcDNA3.1(+)/SjTsp2-A recombinant plasmids were constructed and transformed into E.coli DH5?.Twenty four BALB/c mice were randomly divided into pcDNA3.1(+) /SjTsp2-A group(A),pcDNA3.1(+)/SjGST group(B) and pcDNA3.1(+) group(C).Each mouse was injected through musculus quadriceps femoris by three times(two weeks interval) respectively with 100 ?g pcDNA3.1(+)/SjTsp2-A,pcDNA3.1(+)/SjGST,or pcDNA3.1(+).At two weeks after the final inoculation,mice were each challenged by 40?2 cercariae of S.japonicum.Forty-five days after infection,all mice were sacrificed,the number of worms collected and eggs in liver tisssue was counted.Anti-pcDNA3.1(+)/SjTsp2-A antibody was detected by ELISA and protein expression in quadriceps muscle by immunohistochemical staining.The worm reduction rate(44.4%) and egg reduction rate(28.4%) of group A was higher than those of group B and C(P

Objective To clone and express a membrane protein(Tetraspanin 2) gene of Schistosoma japonicum (SjTsp2). Methods A pair of primers was designed to amplify the SjTsp2 gene which was subcloned into prokaryotic plasmid pET28a(+). The recombinant plasmid was transformed into E. coli BL21(D3) and followed by expression of the protein induced by IPTG. The protein was purified by affinity chromatography and used to immunize BALB/c mice. Dilution of antibody against SjTsp2 was determined by ELISA. The protein was also identified by Western blotting. Results Big loop of SjTsp2-A, 228 bp, was amplified in vitro by PCR. Its deduced amino sequence shared 52% similarity with SmTsp2. The soluble recombinant SjTsp2-A was expressed in the experiment and high dilution antibody against the recombinant (1 ∶ 32 000 in maximum) was produced in immunized mice. SjTsp2-A reacted positively with sera of acute and chronic schistosomiasis patients but not with sera from healthy persons by Western blotting. Conclusion SjTsp2 has been expressed and shows certain antigenicity.

Objective To screen the mimic epitopes of Rh(D)blood group antigens and identify their immunity from phage display peptide library.Methods A twelve mer phage peptide library was biopanned with anti-Rh(D)monoclonal antibody immobilized on plastic surface.After three round panning,thirty-five clones were randomly selected and positive clones were identified by ELISA and cross-reaction,followed by antibody competition inhibition assay and DNA sequencing to obtain the mimic epitopes of Rh (D)blood type antigens.The target phage clones were characterized and the antigenicity was analyzed by Western blot.Results After the third round screening,phages were enriched,and eleven positive clones were obtained.According to sequencing and competition inhibition analysis,the same"-WP-Q-"structure existed in seven of the eleven clones,and they had more than 40%inhibition ratio.The other clones had no same characteristics with low inhibition ratio possibly due to non-specific binding.Western blot analysis indicated that these phage clones could be specifically recognized by the anti-Rh(D)serum and they shared the same antigenicity of Rh(D)protein.Conclusions Rh(D)mimotope of"-WP-Q-"structure is successfully obtained by phage peptide library screening with anti-Rh(D)monoelonal antibody.The results lay the foundation for further exploration of pathogenesis and vaccine development of Rh(D)hemolytic diseases of newborn.

Objective To investigate physiological changes in peri extracorporeal circulation period of patients who underwent cardiac valve replacement surgeries with crystalloid solution mixed with colloidal solutions and pure crystal solution as extracorporeal circulation priming solution, and explore the clinical value and practicability of crystalloid solution as the sole extracorporeal circulation priming solution.Methods A retrospective analysis was performed in 130 patients who underwent cardiac valve replacement surgeries.Pure lactated Ringer's solution liquid and Lactated Ringer's solution mixed with Voluven as the extracorporeal circulation priming solution were used.We respectively compared hematocrit at different time points, postoperative blood routine, liver and kidney function, blood coagulation index, duration of intensive care and trachea cannula in two groups.Results There were no significant differences in ages, preoperative blood routine, kidney function, blood coagulation function, duration of operation, clamping time, bypass time, intensive care, postoperative blood routine, kidney function, blood coagulation function and hematocrit at different time points in two groups (P ＞0.05).However, the hospital day of group which used crystalloid solution as extracorporeal circulation priming solution was significant shorter compared to group which used lactated Ringer's solution mixed with Voluven (P ＜ 0.05).Alanine aminotransferase of group which used crystalloid solution as extracorporeal circulation priming solution was significant higher compared to group which used lactated Ringer's solution mixed with Voluven (P ＜0.01).Conclusions Crystalloid solution as extracorporeal circulation priming solution is safe and economy in cardiopulmonary bypass.Pure crystalloid solution as the sole extracorporeal circulation priming solution can be safely used on patients (New York Heart Association class Ⅱ-Ⅲ) who have normal liver and kidney function before the operation of adult heart valve replacement with cardiopulmonary bypass.

Objective To obtain novel vaccine candidate antigens against Schistosoma japonicum. Methods S. japonicum schistosomula cDNA library was screened by using sera of Microtus fortis that was naturally resistant to schistosomiasis. The positive clones were transformated into Escherichia coli BM25.8, E. coli clones containing the plasmid cultured in LB, and then selected for plasmid extraction, the plasmid DNA was digested by EcoRⅠand Hind Ⅲ, and analysed by agarose gel electrophoresis. The positive clones were also sequenced and the data were analysed through the internet Nucleotide BLAST software of NCBI and Expert Protein Analysis system of GeneRunner and HNN. Results Twelve positive clones were obtained after repeatedly immunoscreening the library and their sizes ranged from 300 bp to 1100 bp. Two novel genes (named as Sj-sMf1 and Sj-sMf2) with complete ORF were obtained. The deduced protein of Sj-sMf1 consisted of 93 amino acids while Sj-sMf2 consisted of 61 amino acids. Sj-sMf1 protein predicted containing one cAMP phosphorylation site and Casein kinase C phosphorylation site, respectively. Sj-sMf1 protein predicted containing one Casein kinase C phosphorylation site and two Protein kinase C phosphorylation sites. Conclusion Two novel genes predictably encoding unknown proteins are obtained from immunoscreening of Schistosoma japonicum schistosomula cDNA library by M. fortis sera.

Objective:To explore the design of an evaluation index system with feasibility and appropriateness, and to conduct cloud model evaluation, in order to provide reference for the high-quality development of public traditional Chinese medicine(TCM) medical institutions in China.Methods:The Delphi method and analytic hierarchy process were used to construct an evaluation index system for public TCM medical institutions and calculate the index weights. Then the cloud model theory was applied to comprehensively evaluate the index system.Results:The index system of TCM medical institutions including 5 primary indicators and 12 secondary indicators of " external governance" module and 9 primary indicators and 62 secondary indicators of " internal management" module was constructed. The cloud model of the index system was(4.608, 1.022, 0.151), and the cloud rank was in the range of " very good" .Conclusions:The indicator system constructed in this study is relatively objective, scientific, and reasonable. While ensuring the accuracy and credibility of the evaluation results, it also adds evaluation information, which has certain guiding significance for promoting the high-quality development of public TCM medical institutions.

Objective : To observe the dynamic expression of recombinant lysyl oxidase like protein 1 ( LOXL1) in the lysine oxidase family in the liver of C57BL/6 mice infected with Schistosomajaponicum and explore its role in hepatic fibrosis． Methods: Mice were infected subcutaneously with cercariae of S．japonicum，and sacrificed with euthanasia in 6，9 and 12 weeks after infection．The sera and liver tissues were collected．The levels of liver fibrosis in mice was dynamically evaluated by HE and Sirius red staining，and the serum transaminases were detected．The dynamic expression levels of collagen type Ⅰ ( Col1) ，LOXL1 and α-smooth muscle actin( α-SMA) in liver tissues were determined respectively by Western blot and qPCR. Finally，the dynamic levels of soluble and insoluble collagens were detected. Results: The result of HE and Sirius red staining showed that hepatic fibrosis levels increased at 6 weeks，peaked at 9 week，and decreased at 12 week in response to S．japonicum infection．Western blot and q-PCR showed that the expression levels of LOXL1，Col1 α1，Col3 α1 and α-SMA was significantly up regulated and reached maximum at the 9th week in response to S．japonicum infection．Soluble collagen protein levels reached maximum at the 9th week．and decreased at 12 week，however insoluble collagen protein levels continued to increase． Conclusion There may be a correlation between LOXL1 and fiber cross-linking in the process of hepatic fibrosis in S．japonicum，and it plays a role in promoting hepatic fibrosis.