原细胞

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。

Small intestinal biopsies showed subepithelial collagen deposition with varying degrees of villous atrophy and varying numbers of intraepithelial lymphocytes. Four patients had previous biopsies showing enteropathic changes without collagen deposition. Seven cases were associated with collagenous colitis and 1 also had features of lymphocytic colitis. Three patients also had collagen deposition in gastric biopsies. One case was associated with lymphocytic gastritis. Celiac disease (CD, gluten-sensitive enteropathy) was documented in 4 patients. Five patients made a clinical improvement with combinations of a gluten-free diet and immunosuppressive therapy. Two patients died of complications of malnutrition and 1 of another illness. Clonal T-cell populations were identified in 5 of 6 cases tested. Four of these patients improved clinically after treatment but 1 has died. Collagenous sprue evolved on a background of CD in 4 cases. There was no history of CD in others and these cases may be the result of a biologic insult other than gluten sensitivity.

小肠活检表现为上皮下胶原沉积，绒毛萎缩程度不一，上皮内淋巴细胞浸润数量不等。4例患者以往曾做过活检，表现为其他肠病改变，并没有胶原沉积。7例患者合并胶原性结肠炎，且1例还有淋巴细胞性结肠炎的特征。3例患者胃活检也发现有胶原沉积。1例患者合并淋巴细胞性胃炎。4例患者有乳糜泻（CD，谷蛋白敏感性肠病）病史。5例患者经无麸质饮食和免疫抑制治疗后，临床症状有所改善。2例患者因营养不良合并另一种疾病而死亡。6例患者做了T细胞受体基因重排，其中5例发现有克隆性T细胞群，这5例中有4例治疗后临床症状加重，且1例死亡。4例患者在CD的基础上病发胶原性口炎性腹泻；其余患者无CD病史，他们的发病也许是生物源性损害，而非谷蛋白敏感。

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。

The cells were cultured under the following serum microenvironment. The primary cells of autoserum group were cultured with autoserum, changing the medium with fetal bovine serum after passage. The primary cells of homogeneity foreign serum group were cultured with homogeneity foreign serum, changing the medium with fetal bovine serum after passage. The primary cells of fetal bovine serum group were cultured with fetal bovine serum, and cultured with fetal bovine serum after passage. The primary cells of Dulbecco's modified Eagle's medium group were cultured with serum-free DMEM, changing the medium with fetal bovine serum after passage.

分别以下列血清微环境培养：自身血清组：分离细胞后原代用含大鼠自身血清的培养基培养，传代后换用含胎牛血清的培养基培养；同种异体血清组：分离细胞后原代用含同种异体血清的培养基培养，以后用胎牛血清培养基培养；胎牛血清组：分离后用含胎牛血清培养基培养，以后一直用含胎牛血清培养基培养；DMEM组：分离细胞后原代用不含血清的DMEM培养基培养，传代后换用含胎牛血清的培养基培养。

The healing rate of wound of control group is obviously higher than the model group.(3) Observe under the microscope: When 3 days after we practice the surgery, HE and observe control group wound have a large number of inflammation nature cell soak, model group wound surface inflammatory cell relatively sparse in Balb/c mouse;The control group can seeⅠtype collagenic and green type collagenic day, model group type be obviously less than the control group; When 7 days after we practice the surgery,HE and bitter acid-staining observe control group peripheral inflammation cell soak, have appeared granulation organize.

3镜下观察：创伤后第3d，HE染色观察对照组伤口有大量炎性细胞浸润，模型组小鼠创面炎症细胞较稀疏，天狼猩红染色对照组可见少量呈黄色的Ⅰ型胶原和呈绿色的Ⅲ型胶原，模型组Ⅲ型胶原要明显少于对照组；创伤后第7d， HE染色观察对照组周边炎性细胞浸润，有肉芽组织出现。

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。

The periostea of both experimental and control side of the mandibular ramus were taken and prepared, 2 of each 5 rabbits in a group were prepared for HE stain detection and 3 for proliferating cell nuclear antigen immunohistochemical detection.Results:1, The newly formed bone was detected on the lateral aspect of mandibular ramus after periosteal distraction. The bone was shaped like a hill. It looked very low and was full of holes at postoperative day 28. With the time of consolidation period lengthened, the newly formed bone matured gradually. X-ray examination showed the new bone shaped like a hill. The average values of new bone height at postoperative days 28,35,42 and 56 were 1.86 + 0.15mm, 2.29 + 0.29mm,3.19 + 0.13mm and 4.70 + 0.45mm. Histological examination of both HE stain and picricacid-fuchsin stain showed the increase in the number of osteoblasts and the change in the orientation of collagen fibers and bone trabecula. There were no significant differences between newly formed bone and original bone on the lateral aspect of mandibular ramus at postoperative day 56 histologically.2 Compared with the control side, the distracted periostea proliferated obviously under the microscope, and the number of periostealcells increased with satiation of cellular nuclear per unit area. The images of PCNA immunohistochemical stain of periosteum showed that the experimental periosteum proliferated obviously after distraction compared with the control side.

结果：骨膜牵张成骨的实验研究南京医科大学硕{学位论文l、骨膜牵张后，可见下领升支外侧的骨皮质上有新骨形成，新骨呈山峰状凸起，术后第28天的新生骨较低平，多孔隙，随着固定时间的延长，新骨逐渐成熟；下领升支前后向切线位X线投照显示新骨呈山峰样隆起；经测量，术后第28、35、42和56天组平均新生骨厚度分别为x.86士0.15mm、2.29士0.29mm、3.19士0.13mm和4.70 土0.45mm；脱钙骨组织的HE染色和不脱钙骨组织的苦味酸一品红染色的组织学观察均显示了新生骨在成骨细胞数量上的增长，以及胶原纤维和骨小梁排列方向上的变化，术后第56天的新生骨在组织学上与原升支骨组织已无明显区别。2、HE染色显示，与对照组相比较，实验侧骨膜增生明显，细胞间排列紧密，单位面积内骨膜细胞数增多，细胞核饱满；骨膜PCNA 免疫组化染色显示，与对照侧相比较，实验侧骨膜在牵张后出现了明显的增生迹象，PCNA阳性细胞分布紧密，单位面积内阳性细胞数较对照组多，靠近骨表面的骨膜中的阳性细胞数更多而且分布更为紧密。

The primary culture chondrocytes were round or polygonal shape, half of the 4th generation turned to spindle shape, most of the 6th passage were long spindle shape; The rough endoplasmic reticula and the mitochondrial were rich and well developed, the ability of proliferation of the 3th generation were stronger than the first one, while the 5th generation was weaker.

原代软骨细胞呈圆形或多角形，第4代有一半变成梭形，第5代绝大部分变为长梭形；原代软骨细胞胞浆内粗面内质网和线粒体丰富，第3代软骨细胞生长增殖能力强于原代，第5代增殖能力明显减弱。

When cultured in three-dimensional matrix, the cells were elongated and migrated into the gel, with a phenotype similar to that in the process of angiogenesis and vasculogenesis in vivo .

在二维平面培养中，原代心脏血管内皮细胞呈铺路石样排列；而在三维胶原培养模式中，内皮细胞呈长梭状形态，并迁入胶原培养介质中，和体内血管新生及血管生成过程中的内皮细胞活化表型相似。

Lugalbanda was a god and shepherd king of Uruk where he was worshipped for over a thousand years.

Lugalbanda 是神和被崇拜了一千年多 Uruk古埃及喜克索王朝国王。

I am coming just now,' and went on perfuming himself with Hunut, then he came and sat.

我来只是现在，'歼灭战perfuming自己与胡努特，那麼，他来到和SAT 。