细胞坏死

One hand mechanical obstruct led to the increase of veinous resistance and the obstacle of microcirculation, the other hand the adhesive PMN was activated in excess, the white blood cells released a lot of enzymes, in which PMN-elastase can decompose the components of cell and many albumens, inclusive of immunoglobulin、alexin and fibrication. These components induced the injury of the pancreatic capillary vessels and cell and lysosome enzy made the tissue protein hydrolyze and produced unsaturated fatty acids, which destroyed the structure and function of cellar membrane. The inflammatory cellar factors activate other immunocytes to produce the injury and necrosis of tissue, which aggravated the pathological injury and led to shock、pyaemia and MODS. So ICAM-1 and LFA-1 played an important role in SAP. Frossard found that the expression of ICAM-1 in the rat model, especially in serum、pancreas and lung. All these showed ICAM-1 is an important factor in AP and concomitant lung injury.

胰腺小叶组织局部血管EC首先被激活，ICAM-1表达升高，与被激活的PMN表面表达的LFA-1相结合，"PMN-EC"相互作用加剧，一方面机械性阻塞毛细血管导致静脉阻力增加、微循环障碍；另一方面粘附的PMN过度吞噬或激活，当白细胞吞噬的颗粒不能被封闭隔离，连同细胞内的酶被释放出来，其中的PMN-elastase能够降解细胞基质中各种成分，水解多种蛋白，加重胰腺的毛细血管内皮细胞和腺泡的损伤；释放的溶酶体酶使组织蛋白水解，产生的不饱和脂肪酸引发脂质过氧化方应，破坏细胞膜的结构和功能；释放的炎性细胞因子，激发其他的免疫细胞的功能，导致进一步的组织损伤和坏死，加重SAP的病理损伤，最终导致休克、脓毒血症及多器官功能障碍等严重后果。

The following methods were used in the study, such as viral inoculation of animal, pathological method, flow cytometry, RT-PCR, in situ hybridization, immunohistochemistry and terminal deoxy nucleotidyl transferase mediated dUTP nick end labling . Morphological changes of the infected chickling, dynamic changes of T, B cells and T subsets, changes of adhesion molecules on the lymphocyte surface, necrosis and apoptosis of lymphocyte and neurons, changes of MIP-1β and IL-8 producing cells in the brain, cell types of perivascular tuffing in cerebral tissue were systematically studied.

在研究过程中，采用了病毒接种技术、普通病理学研究方法、流式细胞仪技术、RT-PCR技术、原位杂交技术、免疫组化技术、凋亡细胞末端标记技术，系统研究了不同日龄的SPF雏鸡人工感染AEV-NH937株后的病理变化，T和B淋巴细胞、T淋巴细胞亚群的动态变化规律，淋巴细胞表面某些粘附分子的变化，淋巴细胞和神经元的坏死与凋亡，雏鸡脑组织中产生趋化因子MIP-1β和IL-8细胞的变化规律，脑组织中围官性细胞浸润的细胞类型。

The following methods were used in the study, such as viral inoculation of animal, pathological method, flow cytometry, RT-PCR, in situ hybridization, immunohistochemistry and terminal deoxy nucleotidyl transferase mediated dUTP nick end labling. Morphological changes of the infected chickling, dynamic changes of T, B cells and T subsets, changes of adhesion molecules on the lymphocyte surface, necrosis and apoptosis of lymphocyte and neurons, changes of MIP-1β and IL-8 producing cells in the brain, cell types of perivascular tuffing in cerebral tissue were systematically studied. The key research results were.(1)The average percentage of CD19+cell in blood, bursa and spleen, after 3, 5 days of inoculation, was significantly Phybridization revealed that the number of IL-8 and MIP-1βproducing cells was increased in the infected brain.

在研究过程中，采用了病毒接种技术、普通病理学研究方法、流式细胞仪技术、RT-PCR技术、原位杂交技术、免疫组化技术、凋亡细胞末端标记技术，系统研究了不同日龄的SPF雏鸡人工感染AEV-NH937株后的病理变化，T和B淋巴细胞、T淋巴细胞亚群的动态变化规律，淋巴细胞表面某些粘附分子的变化，淋巴细胞和神经元的坏死与凋亡，雏鸡脑组织中产生趋化因子MIP-1β和IL-8细胞的变化规律，脑组织中围官性细胞浸润的细胞类型。

Parthenolide is extract from feverfew, it's main component is sesquiterpene, it is commanly used in west country and is mainly used to therapy skin infection, rheumatic disorder, migraine. But recent research indicate that parthenolide can regulate many pathway of signal in the cell, inhibit the cyclooxygenase, TNF-α, interleukin-1 expression induced by lipopolysaccharide in macrophage, inhibit the iNOS expression in human monocyt, inhibit the activity of NF-κB in Hela cell, inhibit the growth of tumor cell and can relieve the myocardial ischemical reperfusion injury and so on. But the invention research about the effection of parthenolide on the VSMC is very rare in the world, so we do some research about the effection of inhibition of parthenpolide on the proliferation of VSMC and the signal transduction mechanism.

小白菊内酯(parthenolide，又名Sesquiterpene lactone)是艾叶菊属黑叶母菊的提取物，主要成分是倍半萜烯内酯，西方国家较为常用，主要用于治疗皮肤感染、风湿病和偏头痛，但近年来发现小白菊内酯能调控细胞内多种信号途径，如抑制巨噬细胞中脂多糖诱导的Cox，肿瘤坏死因子α和白细胞介素-1的表达，能抑制人单核细胞中iNOS的表达，能抑制Hela细胞中NF-κB的活性，抑制肿瘤细胞的生长，同时能减轻心肌缺血再灌注损伤等，但小白菊内酯在VSMC中的干预研究目前国内外非常罕见，为了确切评价小白菊内酯对血管平滑肌细胞增殖的抑制作用及相关的信号转导机制，我们进行了一些研究。

Based on the different permeability of DNA-intercalant dyes YO-PRO-1 and propidium iodide to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4μmol/L YP and 4μg/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively.

根据正常细胞、凋亡细胞和坏死细胞的细胞膜对核酸荧光染料的不同选择通透性，用4μmol/LYO-PRO-1和4μg/ml碘化丙啶(Propidiumiodide，PI)染色96孔板中的细胞样品。分别在485/538和530/590的检测波长下借助荧光分光光度计检测细胞样品孔的YP和PI荧光强度。

Results: IL-8 protein was located in the epithelium of viii. The expression of IL-8 was significantly increased in RSA group than that in the control group. Positive IL-8 cells was shown in decidua of RSA group, and its IL-8 level was higher than that in the control group. By hematoxylin-eosin staining. trophoblastic layer was found to get thinner, cells of trophoblastic layer denatured or necrotized, and turned acidophily, and the fibration of villous axis increased in RSA; decidual cells lost connection, a part of decidual cells appeared cytoclasis and turned more acidophilic, and nuclei disappeared in RSA.

结果：在绒毛组织中，IL-8蛋白定位于绒毛上皮细胞的细胞质内，且病例组的表达明显高于对照组；在蜕膜组织中；病例组蜕膜细胞的胞质内可见IL-8蛋白表达，且高于对照组；H-E染色可见病例组绒毛组织的滋养层变薄，细胞变性甚至坏死、嗜酸性增强，绒毛中轴纤维化程度增强；蜕膜组织中蜕膜细胞失去细胞间连接，部分蜕膜细胞解体、核消失。

However, fully consideration of special structure and physiological function of CRP is critical to investigate whether CRP provide protect effect or pathogenic effect in the progression of atherosclerosis. CRP is a member of pentrxins family, which consists of five identical, noncovalently associated 23-kDa protomers arranged symmetrically around a central pore with a 102 A external diameter. Each protomer has a recognition face with a phosphocholine binding site consisting of two coordinated calcium ions adjacent to a hydrophobic pocket. The opposite face of the pentamer is the effector face, where complement Clq binds and Fc receptors are presumed to bind. A cleft extends from the center of the protomer to the central pore of the pentamer. Both faces ofpentamer form a special bi-phase structure: one face is the ligand recognition phase, which can recognize apoptosis cells and enzyme modified low density lipoprotein in which PCh is exposed.

CRP是一种五聚体蛋白，外径为102 A，由5个相同的单体以非共价键结合，形成双面的环形结构，每个单体的配体识别位点或受体结合位点分别位于五聚体两个平面上，分别组成配体识别相和效应器相，从而构成特殊的二相性结构：一面为配体识别相，含有磷酸胆碱(phosphorycholine，PCh)结合位点，能够使CRP识别在病理条件下暴露出PCh的凋亡细胞、酶修饰低密度脂蛋白(enzyme modified low density lipoprotein，E-LDL)等。E-LDL是天然低密度脂蛋白(native Low density lipoprotein，N-LDL)在多种蛋白酶，如胰蛋白酶、神经氨酸酶、胆固醇酯酶等作用下的代谢产物，其主要特点是结构发生改变(N-LDL大小均匀，平均直径250±30 A，E-LDL大小差异极大，直径为100-2000 A)，暴露磷酸胆碱位点，能够被CRP识别；另一面为效应器相，能够与巨噬细胞、单核细胞表面受体FcγRⅡa结合，介导巨噬细胞、单核细胞识别及吞噬凋亡、坏死细胞作用。

Cells were re-suspended and cultured in a humidified incubator with 5%CO_2 at 37℃.

建立巨噬细胞和死亡细胞的共培养模型在96孔板上种植RAW264.7细胞或小鼠腹腔原代巨噬细胞，分为3组，即对照组、凋亡细胞处理组和坏死细胞处理组。

Results: in contrast with model group, SSM can (1) increase the survival rate of ARF in rats, cut down Scr and BUN at 24hr, improve renal function, slightly surpass Vp in its therapeutic effect;(2) mitigate the extent of renal pathologicchanges, decrease the numbers of renal tubule necroses and casts;(3) protect renal ultrastructure such as microvilli, mitochondria, endoplasmic reticulum of TEC, podocytic process of glomerulus epithelial cell,endothelial cell etc, better than Vp;(4) enhance SOD activity, lower MDA contents in renal cortex;(5) enhance NO contents in serum, reduce ET levels in plasma;(6) evidently cut down TNF- a contents in serum, superior to Vp;(7) antagonize calcium overload in TEC;(8) alleviate renal pathological changes in ARF metaphase (at 3d), advance regeneration and recovery of TEC, restore renal structure in ARF convalescence (at 5d), and better than Vp;(9) increase the expression of TEC PCNA and prepro EGFmRNA in ARF metaphase.

结果：与模型组相比，SSM能(1)提高ARF大鼠的存活率，降低24hr时Scr及BUN，改善肾功能，其疗效略优于维拉帕米；(2)减轻ARF肾脏病变程度，减少24hr肾小管坏死数及管腔内管型数；(3)对肾脏的超微结构如肾小管上皮细胞微绒毛、线粒体、内质网、肾小球上皮细胞足突、内皮细胞等形态结构均有一定的保护作用，其疗效略优于维拉帕米；(4)提高肾皮质SOD活性，降低MDA含量；(5)提高血清NO水平，同时降低血浆ET含量；(6)显著降低血清TNF-α的水平，疗效明显优于维拉帕米；(7)拮抗小管上皮细胞[Ca~(2+)]_i的升高；(8)使ARF中期(第3d时)肾组织病变较同期其它两组明显减轻，并使肾小管细胞再生修复提前，至ARF恢复期(第5d时)肾组织结构恢复重建良好；作用明显优于维拉帕米；(9)促进ARF中期肾小管上皮细胞PCNA的表达及EGF前体mRNA的合成，从而增加EGF的合成释放，而维拉帕米无此作用。

In order to study the effects of pre-heating on the sensitivity of cells to tumor necrosis factor-α, the K562 cells were pre-heated at 40℃ for different times to induce heat shock protein 70(HSP70). To observe the effects of TNF-α concentration on the sensitivity of pre-heated cells, cells were pre-heated for 60 min and treated with TNF-α(50,100,200 and 400U/ml) for 16 hours, and the cells vitalities were detected by MTT.

为探讨预热对肿瘤细胞肿瘤坏死因子α敏感性的影响及其机理，将白血病患者胸腔渗出细胞系(K562细胞)在40℃预热不同时间(0、30、60、90、120、150和180min)，使细胞热应激蛋白70(HSP70)高表达；然后将预热60min细胞和未预热细胞分别暴露于50、100、200和400U/ml的TNF-α16小时，用MTT比色法检测细胞活力。

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。