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RESULTS: The collapse was obvious at weeks 2 and 6 after modeling, with only few granulation tissues and fibrous tissues. No chondrocytes were observed. At weeks 12 and 18, fibrous tissues were found at the defect site, and bone tissue and cartilage proliferation was observed at the bottom and margin. However, it was greatly different from normal cartilage. Moreover, there was still no chondrocytes.

结果:实验兔后腿关节的股骨髁关节面的关节软骨缺损后,在制模后2,6周缺损部位凹陷明显,只有少许肉芽组织和纤维组织生长填充,未见软骨细胞爬行生长,12,18周见缺损部位以纤维组织增生为主,底部及边缘可见骨组织和软骨增生,但是镜下形态仍与正常软骨组织区别较大,表面仍未见软骨细胞爬行生长。

The histopathological findings of the partial ADD included the loss of chon- drocyte, horizontal splitting of the condylar cartilage and synovitis at early stage, and the hyperplasia of chondrocyte, the disarrangement of collagen fibers at late stage, but the articular surface remained intact.

完全性关节盘前移位和关节盘穿孔的组织病理研究发现,髁状突软骨变薄,部分区域软骨丧失,软骨细胞增生成族,肉芽组织形成,软骨及软骨下骨出现水平及垂直裂隙,软骨下骨纤维形成,甚至出现囊肿。

Objective Cartilage can be divided into three types: hyaline cartilage, elastic cartilage and fibrocartilage,according to different proportion of extra-cellular matrix.

目的:根据软骨细胞外基质的成分不同,将软骨分为透明软骨、弹性软骨和纤维软骨三种类型。

After 12 weeks,the result showed the operation area of group A articular surface was smooth,with the surrounding normal cartilage naturally straight flush, transparent form new cartilage tissue;Group B restoration surgery the basic integrity of the cartilage tissue,but the center of many not fully integrated,there is slight depression,surface wear,good subchondral bone formation;control group, depression joint operation areas,non-articular cartilage formation,such as lack of bone joints.

术后12周取材,A组修复区可见部分软骨样组织,关节软骨无磨损,修复的软骨呈白色半透明外观,与周围正常关节软骨有连续性,可见一明显的凹陷,无明显软骨下骨外露;B组修复区也可见部分软骨样组织,关节软骨在修复区可见磨损,软骨呈白色不透明外观,软骨下骨形成较好;空白对照组术区关节凹陷,无关节软骨组织形成,关节下骨缺如。

RESULTS AND CONCLUSION: The seeded cells had chondrogenesis in scaffolds under certain conditions.

结果与结论:种子细胞在三维支架中特定条件下有成软骨效应,骨髓间充质干细胞在含20%透明质酸的胶原/透明质酸支架扩增少于3代有成软骨效应,软骨细胞传代数更高仍然有成软骨效应,但脂肪源性成熟基质细胞成软骨效应能力较弱。

Elastic fibers were well-distributed,and vascular endothelial cell did not immigrate.We suggested that expression of ectogenic VEGF triggered paracrine and autocrine of VEGF of chondrocyte and co-acted with VEGF receptor 2 to enhance permeability of chondrocyte and improve internal construct of engineering cartilage,and prevent vascularize proceed.

转VEGF基因软骨细胞作为组织工程的种子细胞与pluronic F-127复合后可于裸鼠体内形成转基因组织工程软骨,与对照组相比,转VEGF基因组织工程软骨具良好的生物学特性,结构均一且与正常软骨组织相似,软骨ECM的GAG、COLⅡ、COLⅩ增多,RunX2、Sox9表达增高,细胞处于增生期的肥大状态,初步分析其原因可能是转染后外源性的VEGF持续表达触发了软骨细胞VEGF自分泌,并通过VEGFR-2作用于软骨细胞,提高了软骨细胞活性,促进其存活与增殖,但未在软骨组织内引起血管内皮细胞的迁移及小血管形成。

The human cartilages are composed of chondrocyte and extracellular matrix,the form of chondrocytes are hypertrophy and the quantity are less;the ECM of cartilage are compised of type II collagen and proteoglycan. Articular cartilages are all hyaline with little fibers. Trauma and arthritis are the main cause of cartilage injury,the ommilayer injury ofcartilage can be recovered by marrow,but because of without stimulation mechanism,the new tissues are merely fibrocartilages,they can not be coincide with hyaline cartilage in menchanics;the purely damage of articular cartilage can not stimulate chondrocyte to regenerate because of without blood circulation,thus,the plerosis of articular catilage can not depend on the proliferation of local chondrocyte.Ever since,people tried their best to find a way to reconstruct articular cartilage.

造成人体关节软骨损伤的原因主要为创伤和关节炎,关节软骨全层损伤可由于骨髓中间充质干细胞的高速增殖修复,但这种修复由于缺乏相应的刺激机制,只能形成纤维软骨,而不能形成符合关节生理、力学要求的透明软骨;单纯软骨部分损伤软骨组织内无血管,软骨细胞迁移迟缓,无法使损伤区域软骨细胞再生,因此,关节炎及关节创伤后的软骨修复不能依赖于软骨细胞的增殖和迁移。

The human cartilages are composed of chondrocyte and extracellular matrix , the form of chondrocytes are hypertrophy and the quantity are less; the ECM of cartilage are compised of type Ⅱ collagen and proteoglycan. Articular cartilages are all hyaline with little fibers. Trauma and arthritis are the main cause of cartilage injury, the ommilayer injury ofcartilage can be recovered by marrow, but because of without stimulation mechanism, the new tissues are merely fibrocartilages, they can not be coincide with hyaline cartilage in menchanics; the purely damage of articular cartilage can not stimulate chondrocyte to regenerate because of without blood circulation, thus, the plerosis of articular catilage can not depend on the proliferation of local chondrocyte.

造成人体关节软骨损伤的原因主要为创伤和关节炎,关节软骨全层损伤可由于骨髓中间充质干细胞的高速增殖修复,但这种修复由于缺乏相应的刺激机制,只能形成纤维软骨,而不能形成符合关节生理、力学要求的透明软骨;单纯软骨部分损伤软骨组织内无血管,软骨细胞迁移迟缓,无法使损伤区域软骨细胞再生,因此,关节炎及关节创伤后的软骨修复不能依赖于软骨细胞的增殖和迁移。

The human cartilages are composed of chondrocyte and extracellular matrix , the form of chondrocytes are hypertrophy and the quantity are less; the ECM of cartilage are compised of type Ⅱ collagen and proteoglycan. Articular cartilages are all hyaline with little fibers. Trauma and arthritis are the main cause of cartilage injury, the ommilayer injury ofcartilage can be recovered by marrow, but because of without stimulation mechanism, the new tissues are merely fibrocartilages, they can not be coincide with hyaline cartilage in menchanics; the purely damage of articular cartilage can not stimulate chondrocyte to regenerate because of without blood circulation, thus, the plerosis of articular catilage can not depend on the proliferation of local chondrocyte. Ever since, people tried their best to find a way to reconstruct articular cartilage.

中文题名人骨髓基质干细胞成软骨诱导及多孔复合材料作为细胞载体的体外实验研究副题名外文题名 Cartilage induction of human mesenchymal stem cells and experiment on compound porous materials as cells' scaffold in vitro 论文作者刘晓岚导师周江南学科专业外科学研究领域\研究方向学位级别博士学位授予单位中南大学学位授予日期2003 论文页码总数68页关键词骨组织工程软骨细胞骨髓基质干细胞壳聚糖高分子外消旋聚乳酸馆藏号BSLW /2003 /R68 /10 造成人体关节软骨损伤的原因主要为创伤和关节炎,关节软骨全层损伤可由于骨髓中间充质干细胞的高速增殖修复,但这种修复由于缺乏相应的刺激机制,只能形成纤维软骨,而不能形成符合关节生理、力学要求的透明软骨;单纯软骨部分损伤软骨组织内无血管,软骨细胞迁移迟缓,无法使损伤区域软骨细胞再生,因此,关节炎及关节创伤后的软骨修复不能依赖于软骨细胞的增殖和迁移。

Using New Zealand rabbit ears cartilage makes chondrocyte suspension, Fibrinogen was mixed with D-hank's makes a final fibrinogen concentration of 50mg/ml mixture solution, then chondrocytes resuspend with fibrinogen solution, chondrocytes-seeded fibrinogen was mixed with thrombin (25U/ml in 40mM calcium choride) to make chondrocytes/fibrin glue polymer. Pluronic F-127 was mixed with D-hank's makes a final Pluronic F-127 concentration of 400mg/ml mixture solution, then chondrocytes resuspend with Pluronic F-127 solution to make chondrocytes/Pluronic F-127 polymer. The chondrocyte concentrations was 10 million chondrocyte/ml of polymer.

为确定纤维蛋白凝胶与Pluronic F-127在注射方式形成组织工程化软骨过程中的优劣,我们进行了如下实验:应用新西兰大白兔耳软骨获取软骨细胞并体外培养,纤维蛋白原应用D-hank's液配制为50mg/ml,然后以纤维蛋白原溶液重悬软骨细胞,再与40mM的氯化钙配制的25U/ml凝血酶混合形成软骨细胞—纤维蛋白凝胶复合物;Pluronic F-127应用D-hank's液配制为400mg/ml,同样用PluronicF-127溶液重悬软骨细胞而形成软骨细胞—Pluronic F-127凝胶复合物。

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