细胞化学

On the morphological differentiation,by inducing of fetal calf serum,it wasshown that the cytotrophoblast cells underwent aggregation first,and the adjacentcytoplasm membranes became tightly contacted through desomosomes andcytoskeleton.Then the adjacent cytoplasm membranes disintegrated fragmentallyto form fusion pore-like structure.Finally the multinuclear cells formed and inwhich the intercellular desmosomes disappeared.Immunocytochemical stainingfurther demonstrated that the formation of syncytium was through cell membranefusion rather than the amitosis process.

细胞形态分化的研究结果表明：早孕胎盘来源的细胞滋养层细胞在血清诱导下，首先发生聚集，并通过桥粒和细胞骨架使相邻的细胞膜紧密接触；之后细胞膜发生片段断裂，形成融合孔样结构；细胞间桥粒消失，最终形成多核的合体细胞；免疫细胞化学分析进一步证实合体化是通过细胞膜融合而非无丝分裂（amitosis即细胞核分裂而细胞质不分裂）实现的。

And thaw, and cell fusion; Understanding the principles and operation procedures for autoradiograph; Mastering the skills of writing experiment reports and graphic drawing under microscope.

本课程是农业与生物学院本科学生的一门实验课程，本课程主要内容包括：掌握和了解普通光学显微镜、几种特殊显微镜和电镜的一般原理和使用方法；掌握显微镜临时标本的制备，会进行显微测量、细胞计数和染色体观察；初步观察细胞生理现象、细胞化学成分显示、染色体标本制备；掌握细胞培养、细胞冻存与复苏、细胞融合等技术的原理和方法；了解细胞放射自显影技术的原理和操作步骤；掌握实验报告的书写和镜下绘图的技能。

The present study is to culture otocyst cells in fetal, cochlear sensory epithelial cells of neonatal and mature respectively in vitro with refinement of culture media and techniques. Immunocytochemistry and Bromodeoxyuridine labeling are used to detect properties and mitotic status of cultured cells. With the observations about proliferation and differentiation in cultured cells, we want to find the origin of regenerative cells and to understand how these progenitors proliferate and differentiate.

本研究分别对胎鼠听囊、新生鼠听觉上皮和成年鼠听觉上皮用自然培养方法进行体外培养，并应用免疫细胞化学方法，对培养细胞进行染色标记和观察，系统了解哺乳动物听觉细胞发生、发育过程，以及这些细胞在体外的增殖和分化过程，旨在了解哺乳动物听觉细胞听觉上皮感觉细胞的来源，发现细胞生长的前体细胞。

Objective: To investigate the effects of sulindac on the growth and motility of ovarian cancer cell SKOV3 and its involved mechanism. Methods: In order to detect the effects of sulindac on cultured SKOV3, sulindac with different concentration and untreated control were set up. The changes in morphology of SKOV3 were observed by phase contrast microscopy. MTT colorimetric assay was used to study the effects of sulindac on SKOV3 cell growth. Motility of SKOV3 was determined by scarification assay.

目的：研究舒林酸对卵巢癌SKOV3细胞的生长和细胞运动的影响以及对基质金属蛋白酶2（MMP2）、环氧合酶-2(COX-2)表达的影响：方法：将不同浓度的舒林酸加入培养液中观察其对SKOV3细胞形态的影响，采用MTT法观察舒林酸对SKOV3细胞生长的影响，通过划痕试验观察舒林酸对SKOV3细胞运动的影响，利用免疫细胞化学方法研究舒林酸对SKOV3细胞MMP2、COX-2表达的影响。

Methods: in order to detect the effects of sulindac on cultured skov3, sulindac with different concentration and untreated control were set up. the changes in morphology of skov3 were observed by phase contrast microscopy. mtt colorimetric assay was used to study the effects of sulindac on skov3 cell growth. motility of skov3 was determined by scarification assay.

目的：研究舒林酸对卵巢癌skov3细胞的生长和细胞运动的影响以及对基质金属蛋白酶2（mmp2）、环氧合酶-2(cox-2)表达的影响：方法：将不同浓度的舒林酸加入培养液中观察其对skov3细胞形态的影响，采用mtt法观察舒林酸对skov3细胞生长的影响，通过划痕试验观察舒林酸对skov3细胞运动的影响，利用免疫细胞化学方法研究舒林酸对skov3细胞mmp2、cox-2表达的影响。

Light microscope and transmission electron microscopy showed that SMMC-7721 cells induced by SAHA had undergone the restorational alteration in morphology and ultrastructure, which were different from those of nontreated cells but were similar to those of normal cells, and the changes were as follows: the cells turned to be flat and spread; the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular; the number of nucleolus reduced and its volume lessened; euchromatin increased while heterochromatin decreased in nucleus; in the cytoplasm, mitochondria grew in number with relatively consistent structure and well-developed mitochondria cristae; Golgi complex turned to be well-developed and typical; rough endoplasmic reticulum increased. Immunocytochemistry assay showed that the expression of AFP and PCNA were declined significantly. FCM analysis showed SAHA could arrest SMMC-7721 cells in G0/G1 phase, with an accumulation of the cells in G0/G1 phase while a decrease of cells in S phase. Semi-quantitative RT-PCR detection revealed that the expression of p21WAFl mRNA was upregulated remarkably in the cells treated with SAHA.

结果：倒置显微镜和透射电镜观察显示，经SAHA处理的细胞增殖速度显著减慢，细胞体积增大，细胞核较小，形状较为规则，核仁数量减少、体积变小，核内常染色质增多而异染色质减少，核质比例减小，细胞质内线粒体数量增多、线粒体嵴发达，高尔基体较为典型，粗糙型内质网增多，呈现出与正常上皮细胞相似的形态变化；MTT比色法测定结果显示不同浓度(2.5、5.0、7.5、10.0uM)SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0/G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0/G1期；RT-PCR检测结果表明，SAHA作用12h后SMMC-7721细胞中p21WAF1 mRNA的表达即有增加，24h后更为明显。

This experiment aimed to systematically investigate and analyze the ethanol activated rates of mouse oocytes, affected factors of oocytes activation, types and development of parthenotes, and the organization and roles of the cytoskeleton during pronuclear formation and migration and early embryo cleavage by ethanol activation of oocytes, embryo culture in vitro and immunofluorescence cyto-chemistry. The results:(1) The ethanol activation of mouse oocytes showed that the activated and developmental rates of oocytes increased significantly with the increase of ethanol concentration and extension of exposure time, but over concentration and exposure time would result in increased fragment rates significantly. 7% ethanol treated oocytes for 7min was the optimum activated condition.

本实验主要利用卵母细胞的乙醇激活、胚胎体外培养和免疫荧光细胞化学方法对小鼠卵母细胞乙醇激活效率、影响因素、孤雌胚类型、发育、原核形成、迁移及早期胚胎卵裂过程中细胞骨架的组装、作用等进行了系统的研究和分析，结果显示：(1)小鼠卵母细胞的乙醇激活结果表明，随着乙醇浓度的升高和作用时间的延长，卵母细胞激活率和发育率都显著提高，但乙醇浓度过高和作用时间过长会导致卵母细胞碎裂率的显著增加，7%乙醇作用卵母细胞7min 为最佳激活条件。

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。

In our research,Wistar rats and SD rats which were parathyroidectomized were used as donors and recepients respectively.The parathyroid gland of the Wistar rat were allografted into the cerebroventricle and other different sites of the brains in SD rat s tereotaxically. Using methods of testing serum Ca~(2+) concentration,histological examination,histochemical and immunocytochemical methods and electron microscopic observation,we investigated the survival of the grafting tissue,its revascularization, infiltrating of monocytes, changes of microglia and astrocytes, the relationship of the CNS with the immune system, and the mechanism of rejection were investigated.The results as following: I .Effects on PTG after allografting into cerebroventricle and cerebral parenchyma of the brain: Testing serum Ca~(2+) conc.

本研究以Wistar大鼠作为供体，切除甲状旁腺的SD大鼠作为受体，在立体定位下将同种异体的PTG移植到大脑的不同部位，设计了侧脑室-内分泌系统移植的代替途径，比较了侧脑室内移植和脑实质内移植的异同，通过测定血清Ca~(2+)的浓度、组织学检查、组织化学和免疫细胞化学以及电镜观察对移植后PTG组织的存活、血管的再生、单核细胞的浸润、CNS内胶质细胞的变化、对免疫系统的影响及免疫排斥的机制进行了研究，结果如下：一、同种异体PTG侧脑室内移植与脑实质内移植的比较：血清Ca~(2+)的检查发现，PTG侧脑室内移植后第7天，血清Ca~(2+)的浓度恢复到8mg％以上，一直持续到移植后第8-12周；移植后第12周仍有60％移植PTG的动物血清Ca~(2+)维持在正常水平。

PCI2 cells were treated under glucose deprivation as model group. Method Morphological and cytoflowmetric methods (the mitochondrial transmembrane potential; MEF method) were applied to evaluate the protective effect of SAB. The RNA levels of grp75 were measured by RT-PCR and immunocytochemistry were performed to investigate the expression level of PC12 cells under glucose deprivation and treated by SAB.

以PCl2细胞（大鼠肾上腺嗜铬细胞瘤细胞）的无糖培养建立细胞缺糖损伤模型，MTT法检测细胞的存活率；应用流式细胞仪检测亚二倍体细胞的比率及线粒体跨膜电位的改变；应用免疫细胞化学、RT-PCR法研究细胞内应激蛋白grp75的表达变化，激光共聚焦显微镜观察SAB作用下Ca(上标 2+)的变化。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。