染色质的

RESULTSdiabetic rats showed typical symptoms involving elevated serum glucose and weight loss (2)The rats in DM+VaD group reacted slower and made more error numbers in Y-maze than the rats in VaD group did, with a prolonged total reacting time in a whole day.(3)More neurons with serious damages, such as edema, ischemia or pyknosis could be found in DM+VaD group than in VaD group. More disarrangement of cone neurons, obvious glia proliferation, and more neuron apoptosis were found in CA area of hippocampus. CONCLUSIONS (1)2-VO in STZ-induced diabetic rats succeeded in establishing VaD animal models.(2) The cognitive dysfunction induced by VaD rats had been deteriorated by diabetes mellitus.(3)Morphology evidences proved that diabetes mellitus aggravated brain damages induced by VaD.PART II Effect of diabetes mellitus on the cholinergic nervous systemin CA1 area of hippocampus of VaDOBJECTIVE To estimate the role of cholinergic nervous system in hippocampus during the process of cognitive dysfunction aggravated by diabetes mellitus in DM+VaD group. METHODS Observe the changes of ChAT protein by immunohistochemistry stain. Chose three rats randomly from each group at each pointat two week, four week, and eight week, then sepa

经腹腔注射链脲佐菌素诱导慢性实验性糖尿病，1周后永久性结扎双侧颈总动脉(2-VO)制作VaD模型记录术后2周、4周和8周各组大鼠的体重及血糖；应用Y-迷宫检测空间定向学第二军医大学博士论文中…丈摘…要1ia；一色恤~﹁染澎糕行HE染色观察组织病理学孪叱；胶质细胞原纤维酸性蛋白protellZ，Cf冰尸标记观察星形胶质细舱的激活情况、橄声软公点娜育票票众糯黑橄居端粗默孺篇筑价井袱第二部分糖尿病对血管性痴呆大鼠海马c心区胆碱能神经系统的影响探讨海马CAI区胆碱能神经系统在糖尿病加重确D一大鼠认知障碍中所实验动物及分组同第一部分，应用免疫组织化学染色方法现察海马CAI{介狱牛第晕军l袭拔常博粗{论义

E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.Results:Myocardial collagen in model group increased obviously compared with normal group from the results of electron microscope、H.E、Sirus red in supersaturated picric acid solution.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型，随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组，并设正常对照组，用药四周后处死动物通过透射电镜观察心肌细胞超微结构；HE染色、苦味酸天狼星红染色观察心肌胶原数量；放免法测定血清透明质酸的含量；免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Methods:Establish the model of myocardial fibrosis in chronic stage of viral myocarditis by injecting CVB3 into Balb/c rats,divide these rats into model group、Tong xin luo low dose group、Tong xin luo medium dose group、Tong xin luo high dose group and set up normal group,after four weeks of giving medicine execute the animals and observe the ultrastructure of myocardial cell with tramission electron microscope,the quantity of myocardial collagen was observed by staining with H.E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型，随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组，并设正常对照组，用药四周后处死动物通过透射电镜观察心肌细胞超微结构；HE染色、苦味酸天狼星红染色观察心肌胶原数量；放免法测定血清透明质酸的含量；免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Then the content of the interleukin (IL-1) in serum were measured by the method of enzyme-linked assay; From the HE dyeing, the change of morph in the arthrodial was observed by the light microscope; From the AB-PAS dyeing, the change of PG in the cartilage matrix was observed; By all of that the therapeutic efficacy of the medicine wine Yong GuKangNing to the disease of osteoarthritis was estimated, and the mechanisms to the disease was also investigated.

用酶免法测定血清中白细胞介素(IL-1)的含量；常规HE染色，光镜下观察关节软骨结构形态学变化；AB-PAS染色，光镜下观察软骨基质中蛋白聚糖的染色变化；评价永康宁骨康宁对膝骨性关节炎的治疗效果，并探讨其作用机制。

Part one Isolating, culturing, chondroblast induction and evaluation of human MSC in vitroObjective:To examine the property and proliferation ability of primary culturing MSC ,mvestgate the effect of lightly centrifugation and chemical definition medium to induce MSC differentiate into chondroblast.

结果：原代骨髓基质干细胞易于体外培养，增殖旺盛；经轻度离心和化学限定培养基诱导后细胞呈高密度生长，仍保持较旺盛的增殖能力，细胞转化成圆形肥大细胞，甲苯胺蓝染色显示软骨细胞外基质 v的特异性异染，11胶原蛋白免疫细胞化学染色呈强阳性，BT干CR鉴定显示＊胶原mRNA丰富的表达。

Staining extent and intensity were evaluated semiquantitatively and mean values for each parameter were calculated. Immunostaining with D2-40 showed positivity in 100% of skeletal myxoid chondrosarcomas, 96% of enchondromas, 95% of low-grade chondrosarcomas, 80% of chordoid meningiomas, and 75% of chordoid gliomas. Staining with S100 demonstrated diffuse, strong positivity in all (100%) chordoid gliomas, skeletal myxoid chondrosarcomas, low-grade chondrosarcomas, and enchondromas, 94% of chordomas, and 81% of extraskeletal myxoid chondrosarcomas, with focal, moderate staining in 40% of chordoid meningiomas.

我们半定量地评估了这些免疫染色的广度和强度，并且计算了它们各自的平均值。D2-40阳性表达于100%例骨的黏液样软骨肉瘤、96%例内生性软骨瘤、95%例低级别软骨肉瘤、80%例脊索样脑膜瘤和75%例脊索样胶质瘤。S100染色弥漫且强烈地表达于所有的（100%）脊索样胶质瘤、骨的黏液样软骨肉瘤、低级别软骨肉瘤和内生性软骨瘤，94%例脊索瘤，81%例骨外黏液样软骨肉瘤，还有，局灶性、中度表达于40%例黏液样脑膜瘤。

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。

MAIN OUTCOME MEASURES: The morphology of BMSCs was observed by the inverted phase contrast microscope. Expression of BMSC surface antigens was detected by flow cytometer. Osteogenous potential was assessed by alkaline phosphatase staining. Adipogenic potential was evaluated by Oil red O staining. Chondrogenic potential was assessed by toluidine blue staining.

主要观察指标：倒置相差显微镜下观察骨髓间充质干细胞的生长状态，流式细胞仪检测细胞表面标志物的表达，采用碱性磷酸酶染色鉴定成骨能力，以油红O染色鉴定成脂能力，以甲苯胺蓝染色鉴定成软骨能力。

METHODS: The level of intracellular calcium of human microglia grown on coverslip,which was loaded by calcium-probe,Fluo-4,and then treated in various experimental processing,was detected by confocal microscopy with time resolution mode.The binding of gp120 to human microglia was determined with confocal microscopy or flow cytometry after treatment with gp120 and stained with anti-gp120-FITC antibody.Phosphorylation of ERK within human microglia with or without gp120 stimulation was analyzed with confocal microscopy following the direct immuno-staining with anti-phosphorylated ERK antibody.

用钙离子探针Fluo-4标记粘附在盖玻片上的人小胶质细胞，运用共聚焦显微镜以荧光强度为指标实时观察各种条件下的细胞内钙离子水平的变化；用gp120处理并用anti-gp120-FITC进行染色，运用共聚焦显微镜术和流式细胞术分析人小胶质细胞与gp120结合情况；用抗磷酸化ERK 抗体免疫荧光方法进行染色，运用共聚焦显微镜术和流式细胞术进行ERK磷酸化水平分析。

At the third passage, BMSCs showed the typical polar swirl morphology. BMSCs were negative for CD34 and CD45, but positive for CD29 and CD44. Following induction, alkaline phosphatase staining, Oil red staining and toluidine blue staining produced a strong reaction in cells.

第3代骨髓间充质干细胞形态单一均匀，呈典型的极性漩涡状生长，不表达造血前体细胞标志抗原CD34和白细胞标志抗原CD45，表达整合素家族成员CD29和黏附分子CD44，经诱导后碱性磷酸酶染色、油红O染色和甲苯胺蓝染色均呈阳性。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。