染色质组型

The chondrocytes of different generation were observed with light-microscope and transmission electron microscope for cellular growth and ultromicrostructure, with the method of MTT assay for grow curve and proliferation, with alcian blue test for GAG of ECM, with immuocytochemistry and RT-PCR (reverse transcript -polymerase chain reaction)for type Ⅱcollagen, with flow cytometry for cell life cycle ,histochemistry for S-A-β-galand so on by which to identify cataplasia and senescence of chondrocytes cultured in vitro.

对软骨细胞退变老化进行观察，采用相差显微镜观察其生长情况，透射电镜观察细胞结构，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，免疫细胞化学法和RT-PCR方法检测Ⅱ型胶原鉴定软骨细胞，以MTT比色法描绘生长曲线、检测生长状态，组化法检测老化相关β-半乳糖苷酶，流式细胞仪分析细胞周期和增殖指数。3。

The third passage chondrocytes were divided into blank group, different desity PAP groups, different desity glucosaminsalfate groups which were passaged to 4th generation and contrast to the 2nd passage group. The chondrocytes of different groups were detected with the method of histochemistry for S-A-β-gal,and with alcian blue test for the content and constructure of GAG of ECM, immuocytochemistry for type Ⅱcollagen and PCNA, MTT assay for proliferation, RT-PCR for type Ⅱcollagen and Aggrecan, flow cytometry for cell life cycle and proliferation index,by which to observe PAP's function regarding to the appearance and functional status in the process of chondrocyte's cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

The 3rdpassage chondrocytes were divided into blank group, different concentration PAP groups,different concentration glucosaminsalfate groups and were sequently passaged to 4thgeneration. The 2nd passage chondrocytes was contrasted as young cells group. Thechondrocytes of different groups were detected with the methods of histochemistry forS-A-β-gal, and with alcian blue test for the content and constructure of GAG of ECM,immuocytochemistry for typeⅡcollagen and PCNA, MTT assay for proliferation, RT-PCRfor typeⅡcollagen and Aggrecan, flow cytometry for cell life cycle and proliferationindex,by which to observe PAP"s function regarding to the appearance and functional status inthe process of chondrocyte"s cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

Objective: To study osteogenic capability of rabbit bone marrow stromal cells and biological characteristical identification in vitro in order to explore the appropriate methods of osteoblast culture in vitro and to select an idea source of seed cells for bone tissue engineer.

目的：研究兔骨髓基质细胞在体外培养条件下的成骨能力及生物学特性，探讨简便易行的成骨细胞体外培养方法，为组织工程选择理想的种子细胞来源。方法：取新西兰白兔骨髓液经离心后得骨髓单个核细胞，以1× 106／ml的细胞浓度进行培养，经传代培养，选择条件培养液培养(1640完全培养液中加入地塞米松10-8Smol／l，β-甘油磷酸钠10mmol／l，VitC 50μg／ml)作为骨髓基质细胞增殖和成骨的调节因子，通过倒置显微镜、HE染色、扫描电镜、碱磷酸酶(alkaline phosphatase，ALP)测定、Ⅰ型胶原免疫组化染色和四环素荧光钙染色等手段对获得的细胞进行生物学特性研究。

Methods 12 guinea pigs of 20 months with 3 cm×3 cm hair sheared from their backs were divided into control group and experimental group. Rhodosin and deer serum preparation was besmeared on the skin of guinea pigs in experimental group, and cosmetic matrix was besmeared on the skin of animals in control group for 60 d. The distribution of type Ⅲ collagen was observed with Sirius red staining and HE staining. The fission index of the cells of stratum basale, the thickness proportion of stratum corneum to epidermis, the volume density of the collagen fibers and small blood vessel in dermis, the number of fibroblasts in aging skin were meassured with technique of sterology.

背部被剪去3 cm×3 cm毛的12只20月龄豚鼠被随机均分为给药组和对照组，给药组涂抹红鹿制剂，对照组涂抹化妆品基质，连续涂抹60 d；应用天狼猩红、HE染色和体视学技术分别观察和测定老化皮肤中Ⅲ型胶原的分布、表皮基底层细胞的分裂指数、角质层同表皮厚度的比例、真皮胶原纤维和微血管的体密度和成纤维细胞的数目。

Bone marrow-derived mesenchymal stem cells are capable of chondrogenesis, making them a possible source of cells for injectable cartilage tissue engineering. There exist different ideas on the ability of mesenchymal stem cells's chondrogenesis in monolayer culture. Because of this, the effect of adult rabbit's bone marrow-derived mesenchymal stem cells chondrogenesis in monolayer culture was studied. The mesenchymal stem cells was isolated from adult rabbit's bone marrow and monolayer cultured. TGF-β1, Vit-C and Dexamethasone were used. Immunohistochemistry analyses and histological staining of H-E, Methylaniline blue and Alcian blue were performed to identify the expression of collagen type Ⅱ and cartilage associated matrix. The results showed that the induced cells expressed and produced collagen type Ⅱ and cartilage associated matrix. This suggests that the differentiation of adult rabbit's marrow-derived mesenchymal stem cells into chondrocyte in monolayer culture is feasible and may be induced by TGF-β1, Vit-C and Dexamethasone.

骨髓基质干细胞的软骨分化潜能使其可能成为可注射组织工程化软骨研究的种子细胞，为探讨体外培养的骨髓基质干细胞在平面诱导条件下软骨分化的可行性，我们进行下列实验：获取并体外平面培养成体兔骨髓基质干细胞，应用TGF-β〓、Vit-C和地塞米松对其软骨分化诱导，诱导后的骨髓基质干细胞行细胞爬片组织学和Ⅱ型胶原免疫组化，结果证实，诱导后骨髓基质干细胞可分泌Ⅱ型胶原，组织学染色可见类似于软骨细胞，由此证明体外培养的骨髓基质干细胞在平面诱导条件下可以软骨分化，其软骨诱导因子为TGF-β〓、Vit-C和地塞米松。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。