型胶原

• Cell adhesion to surface of the substrate is essential to development of the anchorage - dependent cells . only after adhering to surface followed by spreading can cells develop and proliferate . most synthetic polymers used as orthopaedic matrix substitute present hydrophobicity , which may correlates to the low degree of cell attachment . modification with cell adhesion protein / peptides can be benificial to the cell adhesion on polymers and then affect the cell proliferation and differentiation . cell attachment to substrate is primarily mediated by integrins , a widely expressed family of heterodimeric surface receptors . most extrcellular matrix proteins such as fibronectin , osteopontin , collagen type i , bone sialoprotein and vitronectin contain an arg - gly - asp ( rgd ) sequence which is specific to the fixation of cell membrane receptors like integrin . the main aim of this research is to measure , assess adhesion , proliferation of rabbit marrow stromal cells ( mscs ) on the polymers coated by fibronectin , collagen type i or biotie gen , which includes : ( 1 ) biologic characteristics of rabbit mscs were observed by two types of separating method in primary culture . ( 2 ) adhesion , proliferation and differentiation of mscs cultured on polymers coated with biotiegen were assessed . ( 3 ) also , adhesion , proliferation and differentiation of mscs were assessed on plga film or porous plga substrates coated with fibronectin , or collagen type i respectively . ( 4 ) bone formation was observed on the porous plga substrates coated with collagen type i in vivo . this research aims to give new way to make novel synthetic bone with cell adhesion and high bone induction capabilities
  因此将这些蛋白包被、固定到材料表面，观察骨组织工程种子细胞mscs细胞的粘附、生长特性是本研究的中心环节，并从以下方面进行探讨： ( 1 )采用不同原代细胞分离方法，研究其对mscs细胞的生物学特性影响。 ( 2 )检测基因胜肽胶对mscs细胞粘附、增殖及分化的影响。 ( 3 )分别采用型胶原及纤维粘连蛋白( fibronectin ， fn )包被聚乙醇酸-乳酸共聚物( poly ( 1actide - co - glycolide ) ， plga )膜及多孔块型plga材料，观察细胞在单层或三维培养状态下，型胶原及fn对mscs细胞粘附、增殖及向成骨细胞分化效应及能力。
• It has been confirmed that np30 has sensitizing effect on formation of hepatic egg granulomas in mouse model for hepatic egg granuloma of schistosoma japonicum ; immunization with np30 in kunming mice , c57bl / 6 mice and goats obtained worm reduction of 50 . 46 % , 41 . 67 % and 42 . 78 % , respectively . np30 possesses effects of both anti - fecundity and anti - embryonation immunity on female worms of s . japonicum . moreover , np30 plays a significant down - modulatory role to hepatic granuloma and fibrosis ( the diameter , area and volume of egg granuloma were all significantly less than those of control ; the contents of type i , iii of collagen and fibronectin were also significantly less than those of control )
  已对np30分子进行了较为广泛的研究，应用小鼠日本血吸虫肝肉芽肿模型证实np30对虫卵肉芽肿的形成具有致敏作用；对感染宿主(昆明种小鼠、 c57bl / 6小鼠和山羊)具有较好的免疫保护作用(减虫率分别为50 . 46 、 41 . 67和42 . 78 ) ；用np30主动免疫小鼠具有抗雌虫生殖产卵和抗卵胚发育的双重功效；另外，还对血吸虫病肉芽肿和肝纤维化有明显的负调节作用(虫卵肉芽肿的直径、面积和体积均明显小于对照组，肝组织、型胶原及纤维连接蛋白含量均低于对照组) 。
• In conclusion , the method of enzymolysis combined with hplc to extract cp i from pigskin was proposed . this method showed the adventages of high performance , high selectivity and easily amplification . acoording to the experiments of identifying the physico - chemical property and the biosafety of cp i , it was concluded that cp i had high bioactivity and no toxicity effect , and that cp i had the ability to protect cells from oxidative damnification
  综上所述，利用酶解和高效液相色谱制备相结合的方法可以从猪皮中分离纯理学硕士学位论文: i型胶原蛋白的制备、鉴定及其抗uva氧化损伤作用的初步研究化cpi ，该方法具有高效性，高选择性和易放大性;理化性质鉴定及生物安全性试验表明，该物质为纯活性胶原蛋白，无毒副作用;同时对体外细胞的uva氧化损伤具有较好的保护作用。
• In this paper , how to prepare cp i and identify physico - chemical property had been studied , and uva oxidative damage l929 was perliminary discussed . these datas provided theoretic foundation and scientific proof for further research and development of this product . pigskin was used as material
  本文围绕型胶原蛋白( cp )的制备及其理化性质进行研究，同时在其防止长波紫外线( uva )对细胞的氧化损伤方面进行了初步探索，为cp的进一步研发提供理论基础和科学依据。
• To confirm the approval of recombinant pcdnas - tgfjtf , gene and the results of its transfection , we used immuhistochemical staining ( sabc ) . the department of or a logy 4 multiplication and differentiation of bmscs were observed by flow - cytometry ( tcm ) , transilluminating electron microscope ( tem ) and other methods . the bmscs in controlled group was transfected with pcdna3 only
  采用电泳检测pcdna3 - tgf _ 1构建是否成功；通过tgf _ 1免疫组化染色检测转染是否成功；运用形态学观察、透射电镜( tem ) 、流式细胞仪( fcm ) 、碱性磷酸酶( alp )染色、型胶原免疫组化染色等方法，观察转染后bmscs增殖与分化情况。
• In this experiment , radio - immunoassay and hybridization in situ were applied to observe the insulinotropic activities of glp - 1 ( 7 - 36 ) nh2 and reveal the mechanisms underlying this process . methods : rat pancreases were removed from 3 - 5 day - old sprague - dawley rats and dissected into 0 . 5mm3 segments and islets were isolated by the collagenase digestion method of wangling et al . thoroughly washed islets and suspended in modified rpmi - 1640 medium supplemented with 10 % fetal bovine serum , and added to 50ml cell culture flasks
  方法：胰岛的分离参照王玲等的方法，每次实验取新生3 - 5天sd大鼠，无菌条件下剖腹取出胰腺，剪切为0 . 5mm ~ 3的组织块， v型胶原酶消化30min后，离心洗涤，悬浮于完全培养基，接种入50ml培养瓶，于5 co _ 2 、 95空气条件下培养20h ，转板纯化，接种于96孔培养板培养24h ，按实验要求进行实验。
• Methods the 54th generation of transformed human embryonic tendon cells and artificial composite materials of carbon fibers ( cf ) and polyglycolic ( pga ) were co - cultured in vitro to construct tet . lt was frozen in liquid nitrogen with four kinds of cpa for 2 months . post - thawed quickly and transplanted into hind limbs of nude mice , and repaired the defects of achilles tendon . after 2 , 4 , 6 , 8 , 12 weeks , the morphological , histological , ultrastructure , short tandem repeat loci and immunohistochemistry examination were detected , and biomechanical strength of tet were examined . result tendon cell survived and could secret type i collagen after 12 weeks to transplanted into nude mice . in the group of dmso + raffmose + kh2o4 , vacuole in mitochondrion degraded i tendon cell ranged in order , abundant collagen fibers were found and linked each other and the biomechanical strength was increased as time elapsed . c onclusion dmso + raffmose + kh2o4 could protect tet in deep low temperature
  组织工程肌腱制备完成后在四种抗冻剂保护下液氮冻存2月；快速复温后植入裸鼠以修复跟腱缺损， 2 、 4 、 6 、 8 、 12周后取出，观察形态学、组织学、电镜和免疫组织化学变化，短串联重复位点检测和生物力学变化。结果实验组组织工程肌腱体内植入12周后仍有肌腱细胞存活并分泌型胶原；随着时间延长, 10二甲基亚砜( dmso ) +棉子糖( 30mmol l ) + kh _ 2po _ 4 ( 25mmol l )组线粒体空泡减少，肌腱细胞排列整齐，胶原纤维增粗并连接，抗拉强度增高。
• There was no difference in other biologic characteristic of mscs between the two separation method , such as cell anchorage ratio and clone formation ratio . ( 2 ) plga film presented uniformity frame with no protuberance and fissure under scanning electron microscopy ( sem ) . big aperture with smooth wall and average 400 m i n size running - through each other was observed in porous plga substrate , around the big aperture there were many round micropores about 5 m size . all of the structure were equal and uniform , which satisfied the further research work . ( 3 ) mscs adhesion at earlier time was promoted by biotiegenrafter 3h , cell number was ( 1 . 5 0 . 18 ) 105 in the plga film coated with biotiegen group , which was significantly higher than that in plga film group ( p < 0 . 01 ) and higher than that in coverslip group ( p < 0 . 05 ) , which cell number was ( 1 . 04 0 . 21 ) 105 . after 6h and 12h biotiegen could not promote cell adhesion , and cell proliferation and alkaline phosphatase ( alp ) activity were not promoted dramatically during 9 days . ( 4 ) cell adhesion was promoted by fibronectin or collagen type i
  G ） i型胶原、纤维粘连蛋白促进细胞增殖，细胞接种后3 、 6 、 gd三个检测时间点，实验组细胞均明显高于对照组。与1型胶原相比，纤维粘连蛋白刺激作用更强。 ） i型胶原、纤维粘连蛋白尚能诱导mscs细胞向成骨细胞分化，不仅表达成骨细胞标志物ocn 、 alp 、 opnmrna ，而且碱性磷酸酶活性明显增高，碱性磷酸酶及钙结节7第四军医大学博士学位论文一染色均强阳性， i型胶原组mscs细胞碱性磷酸酶活性较fn组更高，有显著性差异；同时，兔疫组化染色表明，经纤维粘连蛋白作用的mscs1型胶原表达阳性。