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.

Objective: To investigate the expression of uteroglobin-related protein 1(UGRP1) in thyroid cells induced by interleukin-1β(IL-1β) and its correlation with Fas/FasL mediated apoptosis. Methods: Control group, IL-1β group and IL-1β+anti-FasL antibody group were established, rat thyroid cells(FRTL-5 cells) were culturedin vitro. The expression levels of UGRP1 and Fas mRNA of each group were detected by real-time PCR and the apoptosis rate of each group was detected by flow cytometry. Results: Compared with control group, the mRNA expression levels of UGRP1 and Fas in IL-1β group and IL-1β+anti-FasL antibody group increased, and the difference was statistically significant(P<0.05). Compared with control group, the early apoptosis rate of thyroid cells in IL-1β group increased(7.49%±1.91%vs28.46%±3.17%), and the difference was statistically significant(P<0.001). Compared with IL-1β group, the early apoptosis rate of thyroid cells in IL-1β+anti-FasL antibody group decreased(28.46%±3.17%vs19.20%±1.75%), and the difference was statistically significant(P<0.05). Compared with IL-1β group, the expression levels of UGRP1 mRNA(2.22±0.31vs2.66±0.28) and Fas mRNA(2.75±0.18vs3.03±0.16) in IL-1β+anti-FasL antibody group were not significantly different(P>0.05). Conclusion IL-1β induces the high expression of UGRP1 and Fas and apoptosis of thyroid cells, the high expression of UGRP1 is not associated with Fas/FasL mediated apoptosis.

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.

OBJECTIVETo prepare sodium alginate-nanohydroxyapatite composite material and to explore its feasibility as a bone repair material.

METHODSSodium alginate-nanohydroxyapatite composite material was prepared using chemical cross-linking and freeze-drying technology. The composite was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM) and its porosity was measured by liquid displacement method. The fifth passage of bone marrow stromal stem cells (BMSCs) were incubated on the composite material and then growth was observed by inverted microscope and SEM. BMSCs were cultured with liquid extracts of the material, methyl thiazolyl tetrazolium (MTT) assay was used to calculate the relative growth rate (RGR) on 1, 3, 5 d and to evaluate the cytotoxicity. Fresh dog blood was added into the liquid extracts to conduct hemolysis test, the spectrophotometer was used to determine the optical density (OD) and to calculate the hemolysis rate.

RESULTSSodium alginate-nanohydroxyapatite composite material displayed porosity, the porous pore rate was (88.6 +/- 4.5)%. BMSCs showed full stretching and vigorous growth under inverted microscope and SEM. BMSCs cultured with liquid extracts of the material had good activities. The toxicity of composite material was graded as 1. Hemolysis test results showed that the hemolysis rate of the composite material was 1.28%, thus meeting the requirement of medical biomaterials.

CONCLUSIONThe composite material fabricated in this study has high porosity and good biocompatibility.

Alginates ; Biocompatible Materials ; Cells, Cultured ; Glucuronic Acid ; Hexuronic Acids ; Humans ; Mesenchymal Stromal Cells ; Porosity ; Tissue Engineering ; Tissue Scaffolds

Objective To prepare sodium alginate-nanohydroxyapatite composite material and to explore its feasibility as a bone repair material. Methods Sodium alginate-nanohydroxyapatite composite material was prepared using chemical cross-linking and freeze-drying technology. The composite was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM) and its porosity was measured by liquid displacement method. The fifth passage of bone marrow stromal stem cells (BMSCs) were incubated on the composite material and then growth was observed by inverted micros-cope and SEM. BMSCs were cultured with liquid extracts of the material, methyl thiazolyl tetrazolium (MTT) assay was used to calculate the relative growth rate (RGR) on 1, 3, 5 d and to evaluate the cytotoxicity. Fresh dog blood was added into the liquid extracts to conduct hemolysis test, the spectrophotometer was used to determine the optical density (OD) and to calculate the hemolysis rate. Results Sodium alginate-nanohydroxyapatite composite material displayed porosity, the porous pore rate was (88.6±4.5)%. BMSCs showed full stretching and vigorous growth under inverted microscope and SEM. BMSCs cultured with liquid extracts of the material had good activities. The toxicity of composite material was graded as 1. Hemolysis test results showed that the hemolysis rate of the composite material was 1.28%, thus meeting the requirement of medical biomate-rials. Conclusion The composite material fabricated in this study has high porosity and good biocompatibility.

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.

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 obtain genes encoding the novel molecules for diagnosis of schistosomiasis. Methods The cDNA library of Schistosoma japonicum (Sj) schistosomula of day 15 post-infection was screened with positive patients sera,and the inserts of positive clones were sequenced. Then the sequences were compared with all sequences in GenBank database by Internet. The positive clones were analyzed by the software in bioinformatics. Results After three rounds of screening,15 positive clones were obtained,and among them 11 clones had the meaningful sequences,in which 5 genes coding for Sj mitochondrion,1 gene coding for Sj myosin and the others were SjCHGC05315,SjCHGC01371,SjCHGC04782,SjCHGC05166,SjCHGC09769,respectively. Conclusion Immunological screening by using positive patients sera can be used to discover Sj antigen genes with a potential value for diagnosis.

Objective To investigate the morphological features of reproductive system of adult Schistosoma japonicum under an optical microscope.Methods Adult schistosomes were obtained from infected mice with cercariae shedding from Oncomelania snails.The adult worms fixed with 10% formalin,dehydrated,imbedded in paraffin,cut at 3 ?m thick,stained by HE staining and then observed under an optical microscope.Results The reproductive organs of adult Schistosoma japonicum such as testicle,ovary,fallopian tube,vitellarium,yolk duct and hystera were displayed distinctly and typically.Conclusions The morphological features of reproductive system of adult Schistosoma japonicum are distinct and typical by using routine pathological techniques preparing and HE staining,which establishes a morphological foundation for the morphological teaching of schistosomes and reproductive biology research.

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.