染色质的

P1 BMSCs, P3 CCs, and P3 ADAS were implanted into collagen/hyaluronic acid scaffold at different concentrations of 10%, 20%, 50%, 80%, and 100%, and the cells were then three-dimensionally cultured in serum-free culture media. Two weeks later, SO staining and immunohistochemical staining were employed to observe the synthesis between of chondroitin sulfate and type II collagen.

取P1骨髓间充质干细胞，P3软骨细胞和P3脂肪源性成熟基质细胞种植入不同比例(10%，20%，50%，80%，100%)的胶原/透明质酸支架中，在无血清培养液中三维培养。2周后，用SO染色和免疫组织化学染色分析，观察硫酸软骨素和Ⅱ型胶原合成情况。

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。

The present studies showed that two cell populations were found in haemocytes: large cell with high granularity and small cell with low granularity by flow cytometry FCM on light scanttering pattern. Two distinct cell types were identified based on phase contrast microscope: one type of cell was dark and dioptric aberration, while the other was bright and dioptric strong. By Giemsa and H.E staining, cytoplasmic staining were heterogeneous and internal particles were obvious in one type of cell, while cytoplastic staining were homogeneous and internal particles were inexistent in the other type of cell. By transmission electron microscope, we found that the mitochondria, Golgi apparatus organelles were rich and internal particles were obvious in one type of cells, and contrary to the another cells.

流式细胞术光散射图谱显示血细胞被分两类，一类为颗粒度高的大细胞，另外一类为颗粒度低的小细胞；相差显微镜观察显示，血细胞可分为胞体暗、折光性差和胞体明亮、折光性强的两类； Giemsa和H.E染色显示细胞分为胞质染色不均一、胞内颗粒明显和胞质染色均一、胞内颗粒不明显的两类；透射电镜超薄切片观察显示，颗粒明显的细胞胞质内线粒体、高尔基体等细胞器较丰富，颗粒不明显的细胞胞质内细胞器较少；负染结果表明血细胞主要分为表面不光滑、突起明显和细胞表面光滑、突起较不明显的两类。

Results Among the three groups,the children's rib cartilage had the most blood vessels,the most chondrocytes,well-distributed stain of matrixes,and the type Ⅱ collagen was expressed actively and highest in photedensity.The rib cartilage of teenager group had less blood vessels,unhomogeny distributed stain of matrixes,the enlarged and separated cartilage lacunas.The rib cartilage in adult group showed the least blood vessels,the least chondrocytes.the hyalinization of perichondium,the most deposition of calcium salt,and the type II collagen was expressed at the lowest level in photodensity.

结果 儿童组肋软骨膜血管最丰富，软骨基质染色均匀，软骨细胞数目最多，Ⅱ型胶原蛋白表达最活跃，平均积分光密度值最高；青少年组软骨膜内血管减少，软骨基质染色出现明显的不均质状，软骨陷窝体积变大，并呈分隔状，陷窝内软骨细胞数目减少，II型胶原蛋白表达较儿童组减弱；成人组软骨膜血管、细胞成分明显减少，软骨膜内的纤维成分明显玻璃样变，钙盐沉积较青少年组时明显增多，Ⅱ型胶原蛋白表达较青少年组减弱。

Results Microscopically, different degrees of hepatocytic degenerative changes and hyperplasic fibrous tissue and typical false lobule formation could be seen. Timm's staining result showed uneven distributed black granular deposits in the hepatocytes. No specific PAS staining was observed. Utrastructurally, the mitochondria were increased in volume and dramatically different in shape. The number of lysomes were increased.

结果 光镜下肝细胞表现为不同程度的退行性变，胶原纤维增生以及典型假小叶形成；Timm's染色阳性，发现不均匀分布黑色颗粒或团块样物质沉积；PAS染色则普遍缺乏特异性染色；超微结构显示线粒体形态多样，体积增大，溶酶体增多，粗面内质网管腔扩张，附着核糖体的脱颗粒，以及胞质水肿，质膜溶解。

Staining observation on complex mesochondrium of marrow-derived mesenchymal stem cells-TCP: It was found with TB staining that the complex of marrow-derived mesenchymal stem cells-TCP inside the joint cavity obviously had positive staining, which indicated that there were glycosidoprotein base formed.

骨髓基质干细胞-磷酸三钙复合体软骨基质特染观察结果：甲苯胺蓝染色发现，植入关节腔的骨髓基质干细胞-磷酸三钙复合体有明显阳性染色，表明有糖蛋白基质形成。

RESULTS: Light microscope showed that the collagen fibers arranged regularly in AECM with blankness of bone lacunas by HE, Mallory-Heidenhain rapid one-step dyeing and that the region around bone lacunas was stained different degrees of blue-green by Alcian blue dyeing.

结果 HE、Mallory- Heidenhain氏快速一步法染色后镜下可见AECM胶原纤维排列规则，骨陷窝空虚；阿利新蓝染色可见 AECM骨陷窝周边染色呈不同程度蓝绿色；免疫组织化学染色可见 FN和 L N的抗体染色阳性部位在细胞外基质。

After treated in 4% paraformaldehyde,we applied Hematoxylin-Eosin and Nissl staining to exhibit the structure of the retinae and ganglion cells respectively,and also we used immunohistochemical ABC method to display the distribution of GFAP-immunoreactive cells that were specifically classified as the astrocytes.

E 染色以显示视网膜结构，Nissl染色显示神经节细胞，免疫组织化学ABC法染色以显示星形胶质细胞特征性标志物胶质纤维酸性蛋白的阳性反应细胞的分布。

Histologically, the seminiferous tubules contained numerous Sertoli cells and more Sertoli-spermatozoa complexes, accompanied by the depletion of Leydig cells with deeply stained nuclei. Mature spermatozoa were stored up in the epididymis, but only a few in the efferent ducts. In the second place was testicular atrophy（32/120; 26.7%）. The seminiferous tubules showed moderate to severe inactivity of spermiogenesis with evidence of only spermatogonia, spermatocytes and Sertoli cells. The Leydig cells were obviously decreased in numbers associated with decrease of lipid droplets in their cytoplasms. Testicular hypoplasia was the third disorders（22/120; 18.3%）. Only a few spermatogonia and Sertoli cells appeared without any spermiogenesis. The associated changes was decreased in Leydig cells and fibrous hyperplasia in the interstitium.Epididymal stones were sometimes found（12/120; 10%）. Grossly, yellowish-white nodules with various sizes and firm in consistency were observed in the epididymis and the front efferent ducts. Microscopically, the epididymal ducts were dilated with voluminous spermatozoa storage, even showed calcification in severe cases. The deposited calcium salts were stained positively by Von Kossa and Alizarin red methods.Amyloidosis was also detected in 10 roosters（8.3%）. Eosinophilic, homogeneous, amyloid-like substances were deposited mainly in the testicular interstitium and the periphery of blood vessels. These substances showed positive reaction by Congo red staining. Five roosters（4.2%）had Marek's lesions in the testis, epididymis and peripheral nerves with infiltration of pleomorphic lymphocytes. Only one case showed epithelial necrosis of seminiferous tubules accompanied by fibrous proliferation in the interstitium.

结果发现，在总共搜集的120个病例中，其中因年老所导致的产精力不佳为最多，占38例（31.7%），於镜下可见大量精虫黏附於Sertoli cell的表面，并可见Sertoli cell数量明显增多而Leydig cell明显减少，且其细胞核呈现浓染的现象，而在其副睪中仍可见到成熟精虫蓄留於管腔中，但在其输精管内却只有少量精虫存在；其次为睪丸萎缩，占32例（26.7%），镜下可见中度至重度无造精作用，其生精小管中只见到精母细胞、精原细胞及Sertoli cell存在，但Leydig cell数量明显减少且其细胞质内的脂质也明显减少；睪丸发育不全，占22例（18.3%），於生精小管内只见到精母细胞及少量Sertoli cell存在，不见造精细胞分化，於生精小管间质可见Leydig cell减少并伴随结缔组织增生；副睪结石，占12例（10%），肉眼下可在副睪及输精管前段见到黄白色大小不一的结节，触感坚硬，於镜下可见副睪管扩张并有大量成熟精虫蓄积，严重时可见钙化现象，以Von Kossa及茜素红染色均呈阳性反应；类淀粉沉著症，占10例（8.3%），镜下在睪丸间质及血管周围可见粉红均质样的物质沉积，以刚果红染色成阳性反应；马立克病，占5例（4.2%），镜下可在睪丸、副睪实质及周边神经内均可见到嗜碱性大小不一的淋巴样细胞浸润；睪丸坏死，占1例（0.8%），镜下可见生精小管上皮细胞坏死脱落及间质结缔组织增生。

Method］Isolated bMSCs were cultured in vitro. rh Fibroblast Growth Factor 1(rhFGF-1)、rh Transforming Growth Factor-β1(rhTGF-β1)、rh Insulinlike Growth Factor-Ⅰ were utilizated. The proliferation of cells was detected by MTT assay, and the macroscopic histology , HE staining and immunohistochemical examinations were performed to seek the best cell factor. In vivo , to investigate the repair of the articular cartilage, bMSCs combined with fibrin glue and rhTGF-β1、rhIGF-I was compared with control group.

方法］rhFGF1、rhTGF-β1、rhIGF-I单独或联合应用对骨髓基质干细胞进行体外诱导培养，应用常规染色、MTT、免疫组织化学染色的方法筛选诱导骨髓基质干细胞成软骨细胞的最佳细胞因子，并将其与骨髓基质干细胞复合于纤维蛋白凝胶制成凝胶复合物，直接种植到兔膝关节实验性关节软骨缺损处，并与对照组相比较，观察软骨修复效果。

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。