染色法

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。

Methods We reported a case of PXA located in the cerebellum of a young adult in view of its extremely unusual location. Morphologic observation, immunohistochemical staining using DAKO EnVision system , histochemical stains including hematoxylin eosin and Gordon Sweet's reticulin methods were used.

采用免疫组织化学DAKOEnVison法、组织化学网质纤维染色和HE染色及形态学观察的方法，报道 1例极罕见于年轻人小脑的多形性黄色星形细胞瘤，探讨其临床病理特点。

Objetive:To investigate the histopathological features and detect the mutation of JAK2V617F in PV、ET and IMF;to analyze the effects of the stages of myelofibrosis and the mutation on the number of leucocyte in PV and ET patients;Methods: the morphological features of 69 patients who dignosed in the first time were studied by HGF and Gomori stains ; the mutation of JAK2V617F was detected in 38 blood or marrow samples of PV、ET and IMF patients by real-time PCR;the effects of the stages of myelofibrosis and the mutation on leucocytes were analyzed;Results: Different stages of reticulin were observed in 15 PV and 15 ET patiens'marrow sections,and the cytosis、the pleomorphism and the bone trabecula side location of megakaryocytes were observed in all PV、ET and IMF patients;the means of leucocyte in patients with and without JAK2V617F were 20.4×109/Land 11.4×109/L respectively;the P value was 0.367 as comparing the leucocytes among different stages of reticulin increasing.

目的：探讨PV、ET、IMF患者的骨髓病理特点并且检测JAK2V617F突变的发生率，分析骨髓中纤维增生程度以及JAK2V617F对PV、ET患者白细胞数的影响；方法：69例初诊患者的骨髓切片经HGF染色以及Gomori染色后进行形态学研究；并利用real-time PCR法测38例患者的骨髓或者外周血的JAK2V617F突变；分析骨髓切片中纤维组织增生程度以及JAK2突变对患者白细胞数的影响；结果：15例PV和15例ET患者的骨髓切片中可以观察到不同程度的纤维组织增生，所有PV、ET、IMF患者骨髓病理中可见到巨核细胞增多、巨核细胞多形性改变以及巨核细胞定位于骨小梁旁；12例PV、4例ET、4例IMF中检测到JAK2V617F；有突变和无突变患者发病时白细胞数均值分别为20.4×109/L和11.4×109/L，比较不同纤维增生程度的PV、ET患者的白细胞数均值，P值为0.367。

The histological sections, with HE and Masson's trichromic staining made, to evaluate the effectiveness of repair of bony defect. Results There existed new cartilage and new immature bone tissue in the bone substitute in 4 weeks after operation, as well as mass of cartilageous cells,osteoblasts, collagen tissue and more small vessels were seen.

方法高温法制备出磷酸四钙，然后在模拟体内环境下将其与无水磷酸氢钙发生水化固化反应，合成水泥型羟基磷灰石人工骨，将自制的CPC人工骨植入新西兰兔桡骨大段骨缺损模型中，术后4、8、12、16周取材，采用HE染色和Masson三色染色分析评价骨缺损修复效果。

Methods Tetra-calcium phosphate was prepared by high heating, than imitated the internal situation of humanbody, for hydrolization and solidation,to synthesize the hydroxy-calcium phosphate,as artificial bone in sement-type ,then im-planteted them into the large segmental defect of radius of New Silan rabbits model. The implanted materials were taken out at 4th, 8th, 12th, and 16th weeks after implantation. The histological sections, with HE and Masson's trichromic staining made, to evaluate the effectiveness of repair of bony defect.

方法高温法制备出磷酸四钙，然后在模拟体内环境下将其与无水磷酸氢钙发生水化固化反应，合成水泥型羟基磷灰石人工骨，将自制的CPC人工骨植入新西兰兔桡骨大段骨缺损模型中，术后4、8、12、16周取材，采用HE染色和Masson三色染色分析评价骨缺损修复效果。

Materials and methods: The lung preparation of 6 SARS death patient (died in 9,14,20,29,33,38d) and 6 macaque model (killed in 7,12,14,14, 32, 35d)were objects. Pathological changes, collagen fibers, lattice fibers, elastic fibers, collagen I and III in lungs and fine structure changes were studied by routine H.E dyeing, trigeminy dyeing, trinitrophenol- sirius red staining and polarization microscope, electron microscope and image analysis. Expression of Vimentin、 TGFβ_1、 TNF α、 IL-1β and MMP-2 were detected by immunohistochemistry.Results:1. Pathological changes of SARS death patient.

材料与方法：以6例SARS死亡患者(分别于发病后9、14、20、29、33、38d死亡)和6只猕猴实验模型(分别于染毒后7、12、14、14、32、35d活杀取材)肺标本为对象，应用H.E染色、三联染色、苦味酸-天狼猩红-偏振光法、电镜和图像分析等技术，对比性观察SARS肺组织病理变化和纤维化的病理过程、胶原纤维、网状纤维和弹力纤维的变化特点、Ⅰ型与Ⅲ型胶原纤维的数量和分布规律，旨在探讨SARS肺纤维化的病变经历过程及特点；利用免疫组织化学和形态计量学技术检测SARS死亡患者肺脏的Vimentin、TGFβ_1、TNFα、IL-1β和MMP-2等与炎症反应和纤维化相关活性因子，探讨其发病机制。

MATERAIL AND METHODS: The cell proliferation inhibition was measured by (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromid,MTT) colorimetric assay. Morphological changes of EJ cells were observed by fluorescence staining of Hoechst 33258. Cell cycle and apoptosis were analyzed by flow cytometry.

材料与方法：四甲基偶氮唑蓝(3-[4，5-dimethylthiazol-2-yl]-2，5 diphenyl tetrazolium bromid，MTT)法检测EJ细胞增殖活力；荧光染色观察凋亡细胞的形态学变化；流式细胞术分析细胞周期及细胞凋亡；免疫细胞化学染色观察caspase-3 蛋白的表达。

We observed the changes of the proprioceptors in the partly injuried ACL and the xenogeneic graft/artificial graft being used to reconstructing the injuried ACL with gold chlorid staining method and laser copolymerization electron micrography method to observe the morphological changes of the neuromechanism in the ligament tissue,with RT-PCR method to evaluate the proportion and the regeneration of the nerve component and with HRP retrograde tracking technology and electrophysiologic study to judge the functional changes of the neuromechanism in the ligament tissue,respectively.

本研究分别制作兔ACL部分损伤动物模型、异种肌腱移植物重建损伤ACL动物模型和LARS人工韧带重建损伤ACL动物模型，分别于术后不同时间点对损伤ACL/移植物进行氯化金染色光学显微镜检查和PGP9.5免疫荧光染色后激光共聚焦电子显微镜检查观察韧带内神经结构形态学变化，用RT-PCR法检测韧带中GAP-43和PGP 9.5的mRNA表达情况以评价损伤ACL/移植物内神经成分比例及神经再生情况，通过HRP逆行追踪技术和电生理检查(包括体感诱发电位和腘绳肌肌电图)评价损伤ACL/移植物内神经结构的功能情况。

Result:The sterandryl substitution group and the vacation surgery group comparatively castrates the group brain stem dead volume obvious increase; The Bax protein expression masculine cell number expression also increases.

线栓法建立大鼠局灶性脑缺血再灌注损伤模型，TTC染色测脑梗死体积及免疫组化染色检测脑组织Bax的表达。结果：丙酸睾酮替代组和假手术组较去势组脑梗死体积明显增加；Bax蛋白表达阳性细胞数表达也增加。

The quality concentrations of serum E2 were detected by radioimmunoassay. Pituitaries were embedded by paraffin and sectioned continuously. The morphologic figures of pituitary were shown by hematoxylin-eosin stain, the expression of estrogen receptor and progesterone receptor were detected by immunohistochemistry.

术后40 d麻醉后处死动物，放射免疫法检测血清雌二醇质量浓度，垂体组织常规石蜡包埋，连续切片，分别进行苏木精-伊红染色和雌、孕激素受体免疫组织化学染色。

I am accused of being overreligious," she said in her quiet, frank manner,"but that does not prevent me thinking the children very cruel who obstinately commit such suicide.""

客人们在卡罗利娜·埃凯家里，举止就文雅一些，因为卡罗利娜的母亲治家很严厉。

Designed by French fashion house Herm è s, this elegant uniform was manufactured in our home, Hong Kong, and was the first without a hat.

由著名品牌 Herm è s 设计，这件高贵的制服是香港本土制造，是我们第一套不配帽子的制服。