Expression and refolding of OLA Ⅰ protein with peptides derived from sheeppox virus / 生物工程学报

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.

Advances in research on echinococcus shiquicus tapeworm / 中华预防医学杂志

Echinococcosis is an age-old disease that causes serious damage to the animal husbandry and the human health perennially. As a newly discovered species of Echinococus, E. shiquicus has the potential public health significance and could be a potential parasitic zoonosis. In this review, its etiology, life cycle, epidemiology, detection and diagnoses, public health etc. are discussed or summarized. Also, a series of comparisons among E. granulosus, E. multilocularis and E. shiquicus are made.

Analysis of positive results of early screening of women of child-bearing age in six provinces(cities)in southern China / 国际检验医学杂志

Objective To investigate the regional distribution of positive results of early screening for women of child-bearing agein the six provinces of southern China(Guizhou,Guangxi,Yunnan,Sichuan, Chongqing and Hunan).Methods The screening of all were performed by red blood cell related parameters、quantitative analysis of hemoglobin、Osmotic fragility test of erythrocyte incubation(one tube method)、dena-tured hemoglobin inclusion body test among 222 645 peoples reproductive age in the six provinces of southern China,and statistical analysis of abnormal screening in different provinces(cities)was carried out.Results A-mong 222 645 cases in the six provinces(cities),abnormal samples were detected in 32074 cases,and the abnor-mal detection rate was 14.39%.The total positive detection rate from high to low according to the order of Guangxi(24.87%),Sichuan(17.53%),Yunnan(14.76%),Hunan(11.53%),Chongqing(10.69%)and Guizhou(8.61%).Among the six provinces(city),the total α and β thalassemia screening positive rate were 16.37%,17.81%,the specific distribution(α/β):Guangxi(7.06% /5.89%),Sichuan(2.74% /3.02%),Hunan (1.87% /2.32%),Yunnan(1.56% /1.57%),Guizhou(1.85% /3.04%)and Chongqing(1.29% /1.97%).The total α and β thalassemia screening suspected positive rate were 50.56%,3.25%,the specific distribution(α/β):Guangxi(11.59% /0.33%),Sichuan(11.27% /0.50%),(9.58% /2.05%)in Yunnan,Chongqing(7.35% /0.08%),(7.12% /0.22%)of Hunan and Guizhou(3.65% /0.07%).Conclusion The six southern provinces (city)positive screening rate distribution is different,the highest in Guangxi,followed by Sichuan,Guizhou is the lowest;suggestions will flow into the southern city and high malaria area included in in screening of thalas-semia.The four-way experiment has a higher positive rate of screening for thalassemia,and screening out the number of abnormal hemoglobin disease,and has great value of reduce the misdiagnosis.In order to further re-duce the misdiagnosis,suggested that serum iron detection into thalassemia screening.Strengthen the screen-ing efforts for thalassemia in high risk families,reducing the birth rate of thalassemia major,improve popula-tion quality,reduce the economic burden of the family and the country.

Anti-Inflammatory and antiviral effects of water-soluble crude extract from Phragmites australis in vitro

Phragmites australis [P. australis], a world wide distributed wetland grass, is traditionally used as food-making helper and spice in China. The pharmacological effect of this plant is poorly understood. Here, we demonstrated that lipopolysaccharide [LPS]-induced production of inflammatory mediators nitric oxide [NO] and reactive oxygen species [ROS], and the pro-inflammatory cytokines tumor necrosis factor-a [TNF-alpha] and interleukin-lp [IL-1P] in RAW264.7 macrophage were significantly inhibited by the crude extract. The inflammation pertinent signaling extra cellular signal-regulated kinase 1/2 [Erkl/2], P38MAPK, C-Jun and NF-kappaB [NF-KB] activated by LPS could be dramatically inhibited by this extract. It also remarkably inhibited bovine herpes virus type 1 [BoHV-1] replication in MDBK cells. Taken together, here, for the first time we provided P. australis

Identification of the 987P protein receptors for enterotoxigenic Escherichia coli / 生物工程学报

The 987P fimbriae of enterotoxigenic Escherichia coli (ETEC) mediates adhesive interactions with brush border vesicle (BBV) of the intestinal epithelial cells from the neonatal piglets. By adhering to intestinal epithelial cells, producing localized multiplication, the 987P ETEC can progress to mucosal surface colonization and concomitant effective enterotoxin delivery. To identify the receptors for the 987P, BBV proteins from piglet intestinal villous epithelial cells were separated by SDS-PAGE and analyzed by Ligand blot, protein bands with a set of 32-35 kD recognized by the 987P fimbriae were subjected to in gel proteolysis with trypsin. The tryptic fragments were separated by microbore reversed phase HPLC(RP-HPLC), samples shown to contain one major peak by MALDI-MS were submitted to Edman sequencing, three peptides were sequenced successfully and the all of three peptides matched the sequences of human or porcine histone H1 proteins. Porcine histone H1 proteins isolated from both piglet intestinal epithelial cells and BBV demonstrated the same SDS-PAGE migration pattern and 987P-binding properties as the 987P-specific protein receptors from piglet intestinal brush border did. The above results indicated that the 987P protein receptors are piglet BBV-derived Histone H1 proteins.

Evaluation of expanded polytetrafluoroethylene membrane in guided periodontal tissue regeneration with scanning electron microscope / 实用口腔医学杂志

Objective: To evaluate the effects of expanded polytetrafluoroethylene (ePTFE) barrier membrane in guided periodontal tissue regeneration(GTR) . Methods:Nine patients with Ⅱ～Ⅲ furcation lesions were treated by guided tissue regeneration using ePTFE membrane. The membrane was retrieved 5 weeks after operation and examined by scanning electron microscopy (SEM) . Results:Large amount of lymphocytes were found on the external upper surfaces of ePTFE, with a little bacteria. The external lower surface of ePTFE was full of fibrocytes, red blood cells and lymphocytes.Conclusion:ePTFE membrane is biocompatible and can close the primary wound over a barrier which stops the downgrowth of junctional epithelium at the early stage of periodontal tissue repair after operation.

Effects of collagen membrane in the treatment of class Ⅱ-Ⅲ furcation defects / 实用口腔医学杂志

Objective: To evaluate the effects of guided tissue regeneration(GTR) in the treatment of degree Ⅱ-Ⅲ furcation defects with collagen membranes.Methods: 11 sites with Ⅱ-Ⅲ furcation defects were treated with collagen membranes(group of GTR); 10 sites received flap operation(group of FO). Clinical indexes, radiographs and gingival crevicular fluid(GCF) was collected before surgery and 3 months after treatment. Alkaline phosphatase(ALP) in GCF was measured. The data were analyzed statistically. Results: 3 months after surgery, the clinical parameters of PD, PFD and AL were decreased(P0.05). The decreace of the clinical parameters measured in group GTR was more than that in group FO(P

Scanning electronic microscope observation of bone marrow stem cells transfected by IGF-1 gene and cultured onto ostrich true bone ceramic / 实用口腔医学杂志

Objective:To evaluate the biocompatibility of insulin-like growth factor I (IGF-1) gene transfected bone marrow stem cells (MSCs) with ostrich true bone ceramic (OTBC). Methods:Rat MSCs were transfected with IGF-1 gene, and positive clones were selected by G418. The expression of IGF-1 protein in the MSCs was detected by immunocytochemical technique. The IGF-1 transfected MSCs were cultured with OTBC and the morphology of the cells was observed by scanning electronic microscope(SEM) at different time point. Results:Immunohistochemical staining suggested that the IGF-1 protein was expressed in the IGF-1 transfected MSCs. The cells adhered to OTBC and stretched well after 24 h of culture. The IGF-1 transfected MSCs proliferated on the surface of OTBC with culture time.Conclusion:The OTBC has a good biocompatibility with IGF-1 transfected MSCs.

Experimental study of atherosclerosis ⅣChanges of aortic smooth cell proliferation, platelet free calcium and aggregation in atherosclerotic rabbits and the effects of 8501 on these changes / 中国药理学通报

Smooth muscle cell (SMC) proliferation, platelet free calcium level and aggregation of experimental atherosclerotic rabbits were investigated in this study. Aortic SMC ofhyperlipidemic rabbits in vitro showed higher growth activity than did normal rabbit SMC. And also hyperlipidemic serum stimulated SMC to proliferate at a significantly greater rate than control serum. Moreover, the level of platelet free calcium and the platelet aggregation was also higher in hyperlipidemic rabbits, indicating that activitated platelets possibly release more PDGF to act as a stimulator to SMC proliferation and calcium is an important factor to activate platelets. Furthermore, SMC from 8501-treated rabbits appeared lower proliferative rate than thecells from hyperlipidemic rabbits. And serum from those rabbits inhibited SMC proliferation compared with hyperlipidemic serum, the inhibitory effect was even stronger than that of normal serum. It may be relevant to the favorable effects of 8501 to TXA2/PGI2 balance.