细胞原质的

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 Three-dimensional gels of type I collagen containing rabbit keratocytes were incubated in the presence of different concentrations of exotoxin A (0.1,1.0,10 mg·L-1),cultivated for 24 h at 37℃, the change of keratocytes in morphology was observed under the light microscope,and the amount of lactate dehydrogenase was measured by enzyme linked immunosorbent assay.

含兔角膜基质细胞（1×105·mL-1）三维胶原凝胶表面添加提纯的不同浓度绿脓杆菌外毒素A（0.1、1.0和10 mg·L-1）后，放入无血清MEM培养液中37℃培养24 h，采用光学显微镜观察培养的兔角膜基质细胞形态学变化，采用酶联免疫吸附法检测损伤的兔角膜基质细胞释放的乳酸脱氢酶含量。

At the small growth stage, three kinds of the sex steroid receptors usually were localized in the cytoplasm or nuclear membrane of oogonia and early primary oocyte, and localized in the cytoplasm and nucleoplasm of oocytes at the large growth stage and mature stage.

在小生长期，3种受体通常定位在卵原细胞和早期初级卵母细胞的胞质或核膜；在大生长期和成熟期，则定位在胞质和核质。

Staining results of ADSCs: The extracellular matrix Alcian blue staining, Safranin O/Fast Green staining and collagen Ⅱ immunohistochemistry were positive.

脂肪间充质干细胞诱导后的细胞化学染色结果：诱导后脂肪间充质干细胞胞外基质阿尔新兰染色、蕃红O/固绿染色、和Ⅱ型胶原免疫细胞化学着色阳性。

Methods Human PDLC cultured in vitro was collected and seeded on Col, HA, HA/Col scaffolds crosslinked by carbodiimide. The influences of scaffolds on cell adhesion and growth were observed by MTT assay. The growth of human PDLC on scaffolds was observed through inverted phase contrast microscope and scanning electron microscope.

将体外培养的人牙周膜细胞接种到碳化二亚胺交联的胶原、透明质酸及透明质酸/胶原支架上；MTT法检测支架对人牙周膜细胞黏附、生长的影响；并用倒置相差显微镜和扫描电镜观察形态变化。

CD34 hematopoietic stem/progenitor cells reside in the bone marrow in close proximity to the endosteal bone surface, surrounded by osteoblasts, stromal cells, and various extracellular matrix molecules. We used a bioartificial matrix of fibrillar collagen I, the major matrix component of bone, as a scaffold for ex vivo expansion of HSCs.

CD34 的造血干细胞/祖细胞存在于接近骨内膜表面的骨髓中，由成骨细胞、间充质细胞以及各种细胞外基质分子所包绕，我们采用生物人工合成的纤维胶原I

MAIN OUTCOME MEASURES: Morphological changes in primary cultured and subcultured mouse BMSCs were measured. BMSC surface antigen of the second passage of mice (CD105, CD44, CD25, CD34) were determined using flow cytometry. The modified method was assessed to harvest the purity of stem cells. Activity of mouse BMSCs was identified using adipogenic and osteoplastic differentiation. The strength of fluorescent cells following multiple passage was observed.

主要观察指标：原代及传代小鼠骨髓间充质干细胞形态变化；对第2代小鼠骨髓间充质干细胞表面抗原，CD105，CD44，CD25，CD34进行流式细胞仪检测，评价改良方法获取干细胞纯度；通过成脂成骨分化，鉴定小鼠骨髓间充质干细胞的活性；观察多次传代后荧光细胞强度。

The results are unsure for experimental use;The rabbit's corneas that were removed with upper-half of corneal limbal epithelium lamella and erased the center corneal epitheliums were transparent with intact corneal epithelium;In the approach,the corneal and limbal epitheliums were burned with a cotton swab socked in 1 mol/L NaOH,there were 4 rabbits' corneal stroma happened perforation or ulcer and symblepharon,and the other one presented corneal epithelium phenotype.This is an applicable method to create the pathological model of corneal limbal stem cell total deficiency.

结果表明，处理后4周，全周角膜缘上皮板层手术切除，中央角膜上皮层用1 mol/L NaOH擦除的5只试验家兔角膜表面全部血管化、结膜化，未发生睑球粘连，角膜基质胶原纤维完整未见溃疡、穿孔等病变，细胞印迹学检查为结膜表型，可作为实验性角膜缘干细胞移植的病理模型；全周角膜缘上皮板层手术切除，中央角膜上皮用生理盐水擦除的5只试验家兔，有2只为结膜表型，另3只为角膜表型，观察期内结果不稳定；半周角膜缘上皮板层手术切除，中央角膜上皮层用生理盐水擦除的5只试验家兔，角膜表面透明，全部为角膜表型；直接用1 mol/L NaOH擦除角膜缘和中央角膜上皮的试验家兔，有4只角膜基质胶原纤维断裂、溶解，并伴有严重的溃疡、穿孔、睑球粘连等病变，不能用于移植试验，另1只角膜表面透明，未见结膜和新生血管长入，细胞印迹学检查为角膜表型。

Two days after bone marrow MSCs compounded to nano-CS/COL scaffold, bone marrow MSCs presented globular shape and were scattered; Four days later, bone marrow MSCs presented shuttle shape, extended and anchored on the surface of nano-CS/COL by pseudopods; Eight days later, bone marrow MSCs proliferated and fused each other, and they secreted a lot of extracellular matrix, then which covered most material particles.

骨髓基质干细胞复合到纳米壳聚糖-胶原纤维支架后2 d，细胞呈球形散在分布；4 d 后细胞呈梭形，延展爬行且有伪足与材料表面锚靠；8 d 时细胞增殖，相互间融合，并有大量的细胞外基质分泌，大部分材料颗粒被覆盖。

In order to elucidate the regulation mechanism of RU5 region to BFV gene expression, BFV3026 provirus DNA was used to construct the plasmids containing different deletion of R region, which were cotransfected with luc report gene locatied behind the IP promoter to BL12 cells, and luciferase activities was assayed, confirming that U5 region could repress the initiation function of LTR as well as IP. The BFV structure genes with different deletion of R region were cloned closely behind to heterogenous CMV promoter, then transfected to 293T cells, RT activity was performed, testifying the R region was required for BFV pol gene expression, and also the function domain was identified within the 100n.t. sequence at the 5′end.

以牛泡沫病毒(Bovine foamy virus， BFV)中国株BFV3026原病毒DNA为材料，构建R区系列缺失质粒，通过对其转染细胞中RT水平及对缺失质粒与luc报告质粒共转染细胞中萤火虫荧光素酶活性的测定，确立U5区对于BFV3026两类启动子LTR和IP均具有负调控作用；同时将带有不同R区的BFV3026结构基因片段克隆于异源启动子CMV之下，通过对其转染细胞293T中RT酶活性的测定，确立R区对于病毒结构基因pol的表达具有一定的调节作用，并将其功能区域初步界定在R区5′端100bp内。

On the other hand, the more important thing is because the urban housing is a kind of heterogeneity products.

另一方面，更重要的是由于城市住房是一种异质性产品。

Climate histogram is the fall that collects place measure calm value, cent serves as cross axle for a few equal interval, the area that the frequency that the value appears according to place is accumulated and becomes will be determined inside each interval, discharge the graph that rise with post, also be called histogram.

气候直方图是将所收集的降水量测定值，分为几个相等的区间作为横轴，并将各区间内所测定值依所出现的次数累积而成的面积，用柱子排起来的图形，也叫做柱状图。