细胞原质的

Application of aFGF to linear incisions in rat skin produces a transient increase in wound tensile strength accompanied by enhanced cellularity and deposition of collagen.

到在老鼠皮的直线的切开的aFGF的申请生产由胶原质的提高的细胞性和免职拉力的力量伴随了的创伤的瞬时的增加。

When the pollen tube comes into the micropyle and contact one of the synergids, both synergids do not show any sign of degeneration.

在二细胞原胚的顶细胞和基细胞中，均可观察到与合子中形态相似的双亲质体和线粒体。

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。

Result:After removal of the bilateral lower molars, the remodeling and degenerative activities in different layers of the condyles increased, such as vesiculation of endoplasmic reticulum, distortion of mitochondria and increase of lysosome. Furthermore, the sequential arrangement of collagen fibrils was disturbed, the collagen network broke down and the proteoglycan granules decreased.

结果：细胞发生了不同程度的改建及退行性反应，如粗面内质网的扩张，线粒体肿胀，溶酶体增多等；基质中胶原原纤维的有序排列结构消失，胶原网受到破坏，同时蛋白多糖颗粒减少；基质中脂质小体及基质小囊等成分增多；负重区较非负重区的改变更显著。

Methods: A 5ml bone marrow was extracted from the lilac of human volunteers. By Percoll fluid and density gradient centrifugation, the MSC was obtained; after the cells filled the bottom of vessel, subcultured them, when they subculture in third generation, redigested them, 500 R/min centrifugate, alter the completed medium to chemical definition medium, examined the form change and prolifration of cells by invert microscope, toluidine blue stain、immunocytochemical stain and RT-PCR to test the type Ⅱ collagen mRNA and proteoglycan.

取健康成人髂后上棘处骨髓5ml，经percoll液分离后密度梯度离心，〓／ml密度接种培养，观察原代细胞的贴壁、增殖状况，细胞长满瓶底后进行传代培养；传至第三代细胞，重新消化后以500转／分钟轻度离心5分钟，〓／ml接种，改用化学限定培养基代替完全培养基培养，倒置显微镜观察细胞生长情况及形态变化，甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖，免疫细胞化学染色检测ECM中Ⅱ胶原的蛋白合成，RT-PCR鉴定诱导细胞Ⅱ胶原mRNA的表达。

The aim are:lTo examine the proliferation ability and potential chondroblast differentiation ;2To find an ideal condition stimulated BMSC differentiate into chondroblast;3To examine the chondroblast proliferation in porous scaffolds and to explore the interaction between cells and materials;4To examine the release of cytokine in vitro.

取健康成人略后上棘处骨髓 5ml，经 percoll液分离后密度梯度离心，10'砌l密度接种培养，观察原代细胞的贴壁、增殖状况，细胞长满瓶底后进行传代培养；传至第三代细胞，重新消化后以500转／分钟轻度离心 5分钟，10V加 l接种，改用化学限定培养基代替完全培养基培养，倒置显微镜观察细胞生长情况及形态变化，甲苯胺蓝染色观察诱导细胞合成细胞外基质QCM）中的蛋白多糖，免疫细胞化学染色检测ECM中＊胶原的蛋白合成，RT干CR鉴定诱导细胞＊胶原mRNA的表达。

In the normal uterus, Cytokeratins immunolabelling were detected in glandular cell, luminal epithelial cell, Vimentin immunolabelling were detected in stromal cell and endoblastic cell; CK7 immunolabelling were not detected in any tissue of the yak utenus.

研究结果显示：未妊娠时，泛角蛋白在子宫内膜腺上皮细胞、腔上皮细胞内表达，波形蛋白在子宫内膜基质细胞内表达，平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达，牦牛子宫任何部位均不表达角蛋白7；妊娠30天左右时，泛角蛋白在子宫内膜腺上皮细胞、子宫内膜腔上皮细胞、滋养层细胞、内胚层细胞和尿囊细胞内表达，波形蛋白在子宫内膜基质细胞和内胚层细胞内表达，平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达，角蛋白7在尿囊细胞内表达，偶尔在腔上皮细胞的细胞核边缘表达；消化法进行原代培养时，组织经胶原酶消化并通过100目和400目筛网组合可以有效地分离原代子宫内膜基质细胞和子宫内膜腺上皮细胞；分离得到的子宫内膜基质细胞活率达90%以上，并可在体外传代7次以上；分离得到的子宫内膜腺上皮细胞活率可达85%以上，并可在体外传代5次以上；RPMI1640培养基最适合子宫内膜基质细胞和子宫内膜腺上皮细胞的生长，维持子宫内膜基质细胞正常生长的FBS添加量为20%，维持子宫内膜腺上皮细胞正常生长的胎牛血清添加量为30%。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

Objective: To explore the mechanisms resulting in the recurrence of urethral scar which make urethral strictures difficult to be cured, a series experiments were conducted to find potential effective factors involved in urethral scar formation and degradation, including the studies of extracellular matrix component of urethral stricture scar, the characteristics of urethral scar fibroblast, and the effects of urine on urethral fibroblast in vitro, as well as the studies to compare the difference of collagenase activity, type Ⅰ collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues and cultured fibroblasts from normal urethra and strictured urethra respectively, and the studies to investigate the effect of antisense TIMP-1 oligodeoxyonucleotide on cell proliferation and collagenase activity of urethral scar fibroblast.

中文题名尿道瘢痕基础研究副题名胶原酶活性，TIMP-1的表达及其反义基因治疗外文题名 Experimental study on urethral scar-activity of collagenase，expression of TIMP-1，and antisense TIMP-1 gene transfection of urethral scar fibroblast 论文作者黄翔导师杨宇如教授学科专业外科学研究领域\研究方向学位级别博士学位授予单位四川大学学位授予日期2002 论文页码总数104页关键词尿道手术瘢痕胶原酶成纤维细胞尿道瘢痕馆藏号BSLW /2003 /R699 /12 目的：研究尿道瘢痕的细胞外基质的组成，尿道瘢痕成纤维细胞的生物学特性以及尿液对其生长的影响；比较胶原酶活性，金属蛋白酶组织抑制因子-1（TIMP-1）以及Ⅰ型胶原含量在尿道瘢痕和正常尿道组织及体外培养的成纤维细胞中的差异；研究反义TIMP-1寡核苷酸对尿道瘢痕成纤维细胞增殖以及胶原酶活性的影响。

On the termination date, the cultured explants were all examined by Western blot, HE and transmission electron microscope. Our results showed that after 12-days in culture, the cultivation treated with AS-ODN reduced the synthesis of AMBN and had a deformed dental cusp with thinner enamel matrix. Ultrastructure analyses showed that there was hardly any cisternae of the rough endoplasmic reticulum in the ameloblasts at the tip of the cusp of AS-ODN treatedexplants. However, on average the enamel matrix was thinner compared with that in the control group. Furthermore, the collagen fibers in extracellular matrix were found disorganized. These findings seemed to provide a direct experimental evidence that tended to indicate that the arrested AMBN translation in cultured tooth germs might result in the delay of the tooth development.

经用Western蛋白印迹检测表明，所设计的反义核酸对AMBN InRNA具有良好的封闭效果并成功阻断了牙胚对AMBN的表达；在缺乏AMBN情况下，与对照组相比，实验组牙胚在体外可以继续生长发育至钟状晚期，出现成釉细胞和成牙本质细胞的分化，成釉细胞可以分化成为分泌期型成釉细胞，胞浆中缺少合成蛋白质所必需的粗面内质网和高尔基氏体，缺乏溶酶体，表明对蛋白合成和脚的能力降低；实验组牙胚有牙尖形成和基质分泌，但牙尖形态异常，基质形成减少，牙尖周围基质最厚处为O.6卜m，明显薄于对照组的5.spin，基质中胶原纤维粗细不等，排列稀疏， 3 第四军医大学硕士学位论文未见钙化现象，充分证明了AMBN在牙胚发育中参与釉质基质形成和矿化过程，影响胶原纤维和牙本质基质的合成，促进成釉细胞对蛋白质的合成和釉质基质蛋白降解。

The absorption and distribution of chromium were studied in ryeusing nutrient culture technique and pot experiment.

采用不同浓度K2CrO4(0，0.4，0.8和1.2 mmol/L)的Hoagland营养液处理黑麦幼苗，测定铬在黑麦体内的亚细胞分布、铬化学形态及不同部位的积累。

By analyzing theory foundation of mathematical morphology in the digital image processing, researching morphology arithmetic of the binary Image, discussing two basic forms for the least structure element: dilation and erosion.

通过分析数学形态学在图像中的理论基础，研究二值图像的形态分析算法，探讨最小结构元素的两种基本形态：膨胀和腐蚀；分析了数学形态学复杂算法的基本原理，把数学形态学的部分并行处理理念引入到家实际应用中。