组织细胞

The dynamic changes of the number of T or B lymphocytes and the content of immunoglobulin G, IgM, IgA in peripheral blood, the number of CD〓, CD〓 lymphocyte subpopulation, T〓 cells and IgG, IgM, IgA antibody-producing cells, the ratios of CD〓 and CD〓 T cells in immune organs (thymus, bursa of Fabricius and spleen) and immune tissues (cecal tonsils and Harderian glands), the content of IgA, IgM, IgG in local fluid (tear, tracheal fluid, intestinal fluid and bile) of one-day chicks infected with chicken infectious anemia virus and CIAV-infected chicks immunized with Newcastle disease vaccine were determined respectively by immunohistochemistry, immunogold-silves and immunoenzyme staining in order to reveal mechanism of immunopathogenesis of chicks infected with CIAV and influence of immune responses of CIAV-infected chicks to ND vaccine.

中文题名雏鸡感染鸡传染性贫血病毒后免疫变化和对新城疫疫苗的应答及其免疫机理副题名外文题名 Immune changes of chicks infected with chicken infectious anemia virus and immune response and mechanism to Newcastle disease vaccine immunity 论文作者郑世民导师刘忠贵教授学科专业基础兽医学研究领域\研究方向学位级别博士学位授予单位东北农业大学学位授予日期2002 论文页码总数105页关键词鸡传染性贫血病雏鸡新城疫疫苗馆藏号BSLW /2003 /S858 /9 本项研究采用免疫组织化学、免疫胶体金及免疫酶方法，首次对CIAV感染1日龄雏鸡和CIAV感染雏鸡ND疫苗免疫及vNDV攻击后，其外周血液的T细胞、B细胞数量和IgG、IgM、IgA含量；中枢免疫器官胸腺和法氏囊，外周免疫器官脾脏及呼吸道和消化道局部粘膜免疫组织哈德尔腺和盲肠扁桃体的ANAE〓、CD4〓、CD〓T细胞、IgG、IgM、IgA抗体生成细胞数量和CD〓与CD〓T细胞比值以及泪液、气管液、肠液、胆汁的IgA、IgM、IgG含量的动态变化规律和CIAV感染对ND疫苗的免疫保护率等进行了全面系统地研究。

Microscope examination:in group A,at 2 weeks,a few inflamatory cells appeared in the holes with many osteoblasts and ostoid appeared around the drilled holes.At 8 weeks,marrow formed in the drilled holes in group B,at 2 weeks,there were a large amount of osteoblasts in the drilled holes and some ostoid formed around thc drilled holes.At 4 weeks,the drilled holes were full of trabeculae.At 8 weeks,the trabeculae matured with the marrow appeared in the intertrabecular space.

组织学结果：2周时A组钻孔区出现少许炎症细胞，边缘出现较多成骨细胞并有骨组织形成，至8周时，钻孔区内形成骨髓组织，只在边缘形成骨小梁结构。2周时B组钻孔区有大量的成骨细胞，边缘有较多骨组织形成，4周时钻孔区内充满重生骨小梁结构，8周时钻孔区内骨小梁成熟，小梁有骨髓组织填充。

Microscope examination:in group a,at 2 weeks,a few inflamatory cells appeared in the holes with many osteoblasts and ostoid appeared around the drilled holes.at 8 weeks,marrow formed in the drilled holes in group b,at 2 weeks,there were a large amount of osteoblasts in the drilled holes and some ostoid formed around thc drilled holes.at 4 weeks,the drilled holes were full of trabeculae.at 8 weeks,the trabeculae matured with the marrow appeared in the intertrabecular space.

组织学结果：2周时a组钻孔区出现少许炎症细胞，边缘出现较多成骨细胞并有骨组织形成，至8周时，钻孔区内形成骨髓组织，只在边缘形成骨小梁结构。2周时b组钻孔区有大量的成骨细胞，边缘有较多骨组织形成，4周时钻孔区内充满新生骨小梁结构，8周时钻孔区内骨小梁成熟，小梁有骨髓组织填充。［结论］骨髓基质干细胞对兔股骨头缺血性坏死有良好的修复作用。

Bone marrow-derived mesenchymal stem cells are capable of chondrogenesis, making them a possible source of cells for injectable cartilage tissue engineering. There exist different ideas on the ability of mesenchymal stem cells's chondrogenesis in monolayer culture. Because of this, the effect of adult rabbit's bone marrow-derived mesenchymal stem cells chondrogenesis in monolayer culture was studied. The mesenchymal stem cells was isolated from adult rabbit's bone marrow and monolayer cultured. TGF-β1, Vit-C and Dexamethasone were used. Immunohistochemistry analyses and histological staining of H-E, Methylaniline blue and Alcian blue were performed to identify the expression of collagen type Ⅱ and cartilage associated matrix. The results showed that the induced cells expressed and produced collagen type Ⅱ and cartilage associated matrix. This suggests that the differentiation of adult rabbit's marrow-derived mesenchymal stem cells into chondrocyte in monolayer culture is feasible and may be induced by TGF-β1, Vit-C and Dexamethasone.

骨髓基质干细胞的软骨分化潜能使其可能成为可注射组织工程化软骨研究的种子细胞，为探讨体外培养的骨髓基质干细胞在平面诱导条件下软骨分化的可行性，我们进行下列实验：获取并体外平面培养成体兔骨髓基质干细胞，应用TGF-β〓、Vit-C和地塞米松对其软骨分化诱导，诱导后的骨髓基质干细胞行细胞爬片组织学和Ⅱ型胶原免疫组化，结果证实，诱导后骨髓基质干细胞可分泌Ⅱ型胶原，组织学染色可见类似于软骨细胞，由此证明体外培养的骨髓基质干细胞在平面诱导条件下可以软骨分化，其软骨诱导因子为TGF-β〓、Vit-C和地塞米松。

In order to understand these differences on gene transcript level, even to get the genes and their expression related to cell culture, we researched the differences of total mRNA of these tissues by RAPD-PCR, the result showed that the difference between embryo and head of nymphae was much closer than embyo with others, which was accorded to the time order of insect upgrown, and the tissue which is more different from embryo on transcript level ismore diiBcult to be cultured in vitro.

四种组织的培养难易程度和分离出的细胞形态的不同，说明存在着组织表达的差异，为了从转录水平上来认识它们的差异，甚而获得与细胞培养难易相关的基因及其表达产物，本实验还采用了RAPD分析的方法进行了这四种组织：中后期虫卵胚胎、若虫头部组织、成虫的精巢和卵巢的mRNA表达差异研究，结果表明：从发育方向上看，四种组织的表达差异有分化越大差异系数越大的趋势，而在相同条件下则越难培养。

1Cubic millimeter tissue sampling was fixed with the mix of 1% glutaraldehyde and 4% paraform in 4℃.

利用兔后腿软组织损伤、海水浸泡模型，分别于损伤后浸泡于海水中2小时，浸泡后分别进行各种低渗盐水冲洗，分别于干预处理后1h、3h、6h、8h、12h，在损伤周围0.5cm范围内的肌肉组织中取材，0.5cm3损伤组织用4%福尔马林固定，HE染色，光镜下观察损伤组织的细胞浸润情况，用免疫组化法检测受伤肌肉组织凋亡相关基因Caspase3表达情况；1mm3损伤组织用1%戊二醛一%多聚甲醛混合固定液(pH7.4 ) 4℃固定，电镜观察其细胞凋亡及细胞膜、线粒体等结构的变化；生化检测Na+，K+-ATPase活性，了解其组间的差别。

Methods: The fibroblasts companying human hepatoma were primarily cultured with explant culture method, first passaged after they spread to the bottom of the culture bottle, and pured by enzyme digestion and repeated strapping the cells were identificated through observing the morphologic change using invert microscope and detecting expression of vimentin, keratin by iMmunohistochemical method.

应用组织块培养法进行人肝癌伴生成纤维细胞的原代培养，细胞铺满培养瓶底后首次传代，通过胰酶消化法和反复贴壁法进行成纤维细胞的纯化；通过倒置显微镜观察细胞形态、免疫组织化学方法检测细胞膜波形蛋白、角蛋白表达情况，对细胞进行鉴定。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法：(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC，细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构；(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定；(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC，并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况，根据各个细胞克隆受Dox调控表达的程度，选择低背景、高诱导表达AT2R的细胞系，作为双重稳定MSC细胞系；蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况；(4)建立大鼠颈动脉球囊损伤动物模型，将双重稳定MSC在术中导入血管，分别于14 d、28 d进行病理切片，检测可调控AT2R对新生内膜增生的影响；采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变，TUNEL法检测血管组织中细胞凋亡的变化情况。

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。

The Plant tissue raise is the plant sterile culture technology, has the totipotency theory according to the vegetable cell, the use plant body exsomatize organ (for example root, stem, leaf, stem point, flower, fruit and so on), the organization (for example cambium, epidermis, cerebral cortex, marrow department cell, endosperm and so on) or the cell (for example big spore, small spore, somatic cell and so on) as well as the protoplast, in aseptic and suitable artificial culture medium and illumination, temperature and so on under artificial condition, can induce injuries the organization, not to decide the bud, the adventitious root, finally forms the complete adult plant the discipline.

植物组织培养即植物无菌培养技术，是根据植物细胞具有全能性的理论，利用植物体离体的器官(如根、茎、叶、茎尖、花、果实等)、组织（如形成层、表皮、皮层、髓部细胞、胚乳等）或细胞（如大孢子、小孢子、体细胞等）以及原生质体，在无菌和适宜的人工培养基及光照、温度等人工条件下，能诱导出愈伤组织、不定芽、不定根，最后形成完整的植株的学科。

The objective is to subjugate and discourage the people, because that allows the elite to continue to rule unopposed.

其目的是压制和打击人民的积极性，因为这可以让实权派继续统治不会沦为反对派。

GOD,this is the second time you vanquished me!

天啊，这是第二次你打败了我！