成纤维细胞

The histopathologic observations of corneas after penetrating keratoplasty in dogs: Thestructure of wholly transparent graft was the same as that of host cornea apart from the upperdonor-host junction where there was minimal irregularity of the corneal lamellae. However, thecorneal lamellae at the lower donor-host junction were neat. The graft with its central part beingtransparent only showed poor coaptation with recipient, in which the graft shifted inward and had marked wrinkles and folds in Descemet"s membrane. On the back of Descemet"s membranethere was hyperplastic stromal fibers that thickened the graft and partially adhered to iris.

5犬穿透性角膜移植术后角膜病理组织学观察完全透明的植片与植床结构一致，仅两者对合处上部纤维紊乱，而下方纤维排列整齐；中央透明的植片与植床对合不良，植片向植床方向嵌入，后弹力层皱缩、折叠，植片下方因有来自植床的基质纤维而增厚，部分对合处与虹膜粘连；浑浊的植片有比较完整的上皮层，植片约2／3深度的基质丧失原来纤维整齐排列结构，代以大量成纤维细胞增生。

Results:(1)NSCs form typical neurospheres under adequate concentration in vitro, which are immunoreactive to Vimentin. Typically and terminally differentiated mature neural cells could not be found without the stimulus of mitogen or only under NSCs self-regulation and self-induction;(2)NSCs derived from hippocampus maintain the character of stem cells much longer with better biological behavior; NSCs passed to the 2-3 passage are the best to graft since they have not differentiated;(3)NSCs cultured in vitro could self-regulate and differentiate into neurospheres and progenitors positively immunoreactive to specific antibodies representing neurons, astrocytes, oligodendrocytes and Schwann cells;(4)There are widespread synaptic contacts between various kinds of descendent clones and cells;(5)Neurospheres could be formed without the stimulus of mitogen when NSCs and OECs are cocultured. Many neurospheres and cells immunoreactive to Vimentin, GFAP, MAP2, 02, p75NGFR, GFAP, S-100, Synaptosis, Vimentin, Tau (Tau is only positive in cocultureof HNSCs+HOECs) could be found;(6)The supernatant fluid triturated from adult rat spinal cord stimulates NSCs to differentiate into neurons, but do not terminally differentiate;(7)Fibroblasts and O4 oligodendrocytes are not supported to grow under this culture medium.Part II: Isolation, culture and identification of rat and human olfactory ensheathing cellsOlfactory ensheathing cells/glials are the most powerful cells to enable the regeneration of axons in the central nervous system.

结果表明：①在适宜的浓度体外培养条件下，NSCs能形成典型的神经干细胞克隆球，Vimentin免疫荧光染色阳性，单靠丝裂原刺激或NSCs自我调节和分化诱导，不会产生典型的终末分化的成熟神经细胞；②海马源性的NSCs维持干细胞特性的时间更长，生物学特性更优；③传至第2～3代的NSCs尚未分化时移植最佳；④体外培养的NSCs能自我调控分化为神经元、星形胶质细胞、O2少突胶质细胞、雪旺氏细胞染色阳性克隆球和前体细胞；⑤各种子代克隆球和细胞存在广泛的突触联系；⑥NSCs与OECs联合培养时，不需丝裂原刺激即能形成克隆球，获得大量Vimentin、GFAP、MAP2、O2、p75NGFR、GFAP、S-100、Synaptosis、Vimentin、Tau(Tau只有人HNSCs+HOECs联合培养时出现阳染)染色阳性的克隆球和细胞；⑦脊髓研磨后的上清液刺激神经干细胞向神经元方向分化，但并不出现终末分化；⑧本研究培养条件不利于成纤维细胞、O4生长。

Results 6 patients with PAP presented with ground-glass opacification of the bilateral lungs on HRCT. Typical chest HRCT scan showed "map-like" lesions in 3 cases and "cobble stone-like" lesions in 3 cases. Multifocal consolidative opacities in the lungs were seen on HRCT in 3 cases. Lung biopsies from 6 patients with PAP revealed a prominent positive periodic acid-Schiff intra-alveolar exudates. There were normal alveolar spetasis in 3 patients while interstitial thickening, fibroblast hyperplasia, interstitial fibrosis in the others.

结果 6例患者在不同层面可见磨玻璃影，其中2例可见小结节影；3例表现为两肺斑片状磨玻璃影与周围肺组织分界清楚，呈地图样改变；3例因磨玻璃影与小叶间隔增厚交织成铺路石样改变；3例可见肺泡实变融合成密度较高的斑片状阴影，1例在肺泡实变区可见&空气支气管征&。6例患者活检肺组织在光镜下显示肺泡腔内充满大量块状或颗粒状嗜伊红物质，PAS染色阳性，AB染色阴性，其中3例肺泡间隔正常，3例肺泡间隔增宽，可见慢性炎性细胞浸润、成纤维细胞增生和胶原沉积。

When reached logarithm growth phase, it was poured or the medium was removed. 5 mL medium containing 10 mg/L mitocin-C was added , kept at 37 ℃, and then cultured in saturated humidity warm box with the CO2 of 0.05 fraction volume for 2 or 3 hours. It was coated with 0.1% gelatin in 6-hole plate, laid for over 30 minutes, and then threw away or the medium with mitocin-C was removed, washed with phosptat buffer for 5 times so as to remove the mitocin-C. 2 mL 0.05% trypsin digestive cells were added, and it was observed under microscope. When crevice appeared, cells became round (about 2－4 minutes), digestion was stopped by adding medium of the same volume, and blew up with straws repetitively to make it into monoplast suspension. Special-used cover glass was put in the center of cell counting chamber. Cells were sucked in by glass siphon, and cell suspension flew out at bucket of up or down-sides of counting chamber, until the cover glass was filled with fluid. Living cell were inoculated at concentrations of 3×108, 5×108, 1×109 L－1 after their number counted.

选取2～5代的小鼠胚胎成纤维细胞，待其至对数生长期倒掉或吸掉培养基，加入含10 mg/L丝裂霉素C的细胞全培养液5 mL，置37 ℃，体积分数0.05的CO2饱和湿度温箱中培养二三小时，在6孔板中加入0.1%明胶包被，放置30 min以上，倒掉或吸掉含丝裂霉素C的培养基，磷酸盐缓冲液清洗5遍，尽量除去丝裂霉素C，加入2 mL 0.05%的胰蛋白酶消化细胞，镜下观察，当细胞间出现裂隙，细胞变圆时（约2～4 min），立即加入等量的全培养液终止消化，并用吸管反复吹打，使之成为单细胞悬液，在细胞计数板中央放置专用的盖玻片，用玻璃虹吸管吸取细胞，让虹吸管在盖玻片上或下侧的计数板凹槽处流出悬液，至盖玻片下被液体充满为止。

Many factors may be involve in the course. To investigate the regulation activity of mesenchymal cells to differentiation of epithelial cells from hair follicle and to study its differentiation property, mesenchymal cells gel was made by nubby dermal papilla cells, free dermal papilla cells, skin fibroblasts. Skin keratinocytes and epithelial cells from hair follicle were inoculated on the gel surface and cultured in air-liquid interface. Three-dimensional model of DPC using to induce epithelial cells differentiation is built in vitro.

为了进一步研究毛囊细胞间的相互作用，探讨毛囊间质细胞对毛囊上皮细胞分化的调节作用，研究毛囊上皮细胞的分化特性，我们利用团块状的毛乳头细胞，游离分散的毛乳头细胞或皮肤成纤维细胞制成间质细胞胶原凝胶，表面接种皮肤角质形成细胞或毛囊上皮细胞，进行气-液界面培养，在体外建立了毛乳头细胞诱导毛囊上皮细胞分化的立体模型。

Skin keratinocytes and epithelial cells from hair follicle organized into epidermoid cyst-like spheroids when cultured on nubby dermal papilla cells gel and epidermoid layer-like structure was formed when they were cultured on free dermal papilla cells and skin fibroblasts gel.

团块状的毛乳头细胞诱导毛囊上皮细胞和皮肤角质形成细胞形成球形结构；而游离分散的毛乳头细胞和皮肤成纤维细胞诱导毛囊上皮细胞和皮肤角质形成细胞形成表皮样层化结构。

Study the bioactivity of the n-HA/PA66 composite and the effects it would be to body's metabolism of calcium and phosphorus ion in vivo.(3) Study the osteo-conductivity and the ability to repair bone defect of the porous n-HA/PA66 composite and the feasibility use it as the scaffold of bone tissue engineering. Objects and Methods as follows: 1.To evaluate the biocompatability of nano-hydroxyapatite crystals and polyamide composite (n-HA/PA66) with the L929 cells.To proceed the morphological observation and take pictures of L929 cells after 1d,2d,4d,and 7d of co-cultured with extract of n-HA/PA66 ,and direct contact with n-HA/PA66.To determine light absorbtion value of every hole under 500 nm with enzyme linked immunity instrument after 1 d,2 d,4 d,and 7 d of contact of n-HA/PA66 extract with L929 cells,and direct contact with n-HA/PA66.In the meanwhile calculate the relative multiplication rate of cells,and evaluate them by six degree tests for cytotoxicity. To investigate the acute and chronic toxic reaction on the whole body induced by the new nano-hydroapatite crystals and polyamide composite(n-HA/PA66)after implanting in vivo and its effects on partial constitution of animal organs after implanting in vivo,and evaluate the potential and degree of subcuticular stimulation reaction.

本实验主要由以下三部分组成：一、n-HA/PA66 复合材料在动物体内、体外的生物相容性及生物安全性评价二、n-HA/PA66 复合材料植入动物体内的生物活性及近期对机体钙、磷代谢影响的实验研究三、网孔 n-HA/PA66 复合材料作为支架修复兔桡骨节段缺损的动物实验研究主要研究目标及方法如下：参照 GB/T16886.5-1997-ISO 10993-5：1992《医疗器械生物学评价细胞毒性试验体外法》之评价标准和要求，采用规定的 L929 细胞(小鼠结缔组织成纤维细胞)，分别经直接接触和材料浸提液与细胞共培养等方式对 n-HA/PA66 复合材料进行细胞毒性测试，采用细胞形态观察法观察两种细胞各组在 24h、48h、72h、5 天后各时相点的细胞形态学变化，并在显微镜下照相，从而对细胞与材料的生物相容性进行定性评价；同时采用细胞生长抑制法，以酶标仪定量测定评价各组 1，2，4，7 天 L929 细胞的相对增殖率，以定量测定并判别材料对细胞的毒性程度。

Methods: Myoblasts isolated from rat muscle samples were cultured and purified clonally in vitro. The 3rd passage cells were incubated with medium including basic fibroblast growth factor and dimethyl sulfoxid.

体外分离培养大鼠成肌细胞并克隆纯化，培养至第3代时加入含有碱性成纤维细胞生长因子和二甲基亚砜的培养液诱导分化，进行RT-PCR分析和免疫荧光细胞化学染色鉴定。

RESULTS AND CONCLUSION: Osteoblast was fusiformed-shaped and had plentiful processes. Nucleus was orbicular-ovate and leaning to lateral side. Soma was large, and plasma was abundant. Alkaline phosphatase staining suggested that a great number of gray-black particles were observed in plasma, and some region was darkly stained.

结果与结论：成骨细胞呈梭形，多突起；细胞核呈卵圆形，偏于一侧；胞体大，胞浆丰富；碱性磷酸酶染色可见胞浆中含有大量的灰黑色颗粒，某些部位染成黑色，定量分析细胞内的碱性磷酸酶、骨钙素水平明显高于成纤维细胞；细胞免疫组织化学染色表明其主要合成Ⅰ型胶原。

RESULTS: Wound healing rate, wound healing time, histopathology analysis, quantity assay of macrophage, determination of hydroxyproline, proliferation of cell, assay of DNA contents and circle of cells, level of transforming growth factor-alpha, levels of interleukin-1, interleukin-6 and tumor necrosis factor, assay of keratinocyte collagenase-1, level of fibroblast growth factor receptor-1, level of monocyte chemoattractant protein-1 and level of keratinocyte plasminogen activator inhabitor type 2 were selected as the evaluation criteria of wound healing.

结果 筛选了创面愈合率、创面愈合时间、组织病理学分析、巨噬细胞定量分析、羟脯氨酸含量测定、细胞增殖情况、细胞DNA含量和细胞周期分析、转化生长因子-α水平、白细胞介素-1、白细胞介素-6和肿瘤坏死因子水平、角质细胞胶原酶-1含量测定、成纤维细胞生长因子受体-1水平、单核细胞化学诱导蛋白-1水平和角质细胞纤溶酶原活化抑制剂-2水平等十三种创面愈合评价指标。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。