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Results:Whith the time and doses of ART incubation extended, ART significantly inhibited the proliferation of SGC-7901 cells and the inhibited effect shows dose-and -time-dependent. Obvious changes of apoptotic morphology were observed by invert microscope, fluorescence microscope, scanning electron microscope and transmission electron microscope, they showed that cells had marked nuclear condensation or the fragmentation of chromation as well as apoptotic , mitochondrial edema and vesicle , with the time of incubation extended , the proliferation rate become slow and the volume became small and transformed. FCM assay indicated that most of the cells were arrested in Go/Gi and the apoptotic peak appeared. During the prolong of incubation time, the apoptosis rate was increased. At the same time, the number of S and G2/M phase cells were decreased. The result of TUNEL indicant that there are apoptosis and necrosis.

结果:随着药物浓度的增加和作用时间的延长,蒿甲醚对胃癌SGC-7901细胞的抑制作用呈时间和浓度依赖关系:在倒置显微镜、荧光显微镜、扫描电镜和透射电镜下可观察到典型的凋亡细胞的形态改变,表现为:核固缩或染色质边聚或凝聚成大块状,可见凋亡小体,线粒体肿胀增殖,严重的空泡化,随作用时间的延长,细胞增殖速度减慢,细胞体积缩小变形;流式细胞术显示胃癌SGC-7901细胞出现明显的凋亡峰,随着作用时间的延长,其凋亡率逐渐升高,细胞周期阻滞在G_0/G_1期,S及G_2/M期细胞数大量减少;流式细胞术TUNEL检测结果显示细胞凋亡和坏死同时存在。

After treated with the samples, the MGC-803 cells are dyed with the fluorescence dye Hoechst 33258. Observed by the fluorescence microscope, the dense dyeing granulometric and nubby blue fluorescence can be seen in the nucleus of MGC-803 treated with AK-9 and AK-11, the nucleus of some cells smash to small fragments.

采用荧光染料Hoechst 33258对经初筛活性样品处理的MGC-803细胞染色,荧光显微镜观察,发现经AK-9及AK-11发酵液处理的细胞,其细胞核内可见浓染致密的颗粒或块状荧光,细胞核发生固缩、碎裂,而对照细胞的细胞核呈均匀弥散荧光,表明这2株菌株可能产生具有诱导细胞凋亡的活性物质。

Most patients' death arise from the metastatic spread of primary tumors There is an urgent need for the finding of novel anti-tumor and anti–metastatic drugs to improve the quality of life and survival rate of patents Justicia procumbens is an herb and used to reduce fever inflammation and canker in Taiwan We've detected that 0 3 μM JA could effectively inhibit the proliferation of oral cancer cells OC-2 and HSC3 but not normal oral keratinocytes in MTS assay JA-treated OC2 underwent apoptosis as suggested by increased sub-G1 DNA contents condensed nuclear morphologh and DNA fragmentiation The migration activity of OC-2 was also significantly reduced by treatment with low concentration of JA The expression of ?

大部分的口腔癌病患死於原位癌的转移和扩散,寻找有效抗癌和抗转移的药物疗法来增进病患的生活品质和存活率,对治疗口腔癌而言非常重要。爵床草是台湾民间常用草药,常被用来消炎止热和治疗口疮、乳瘤。我们利用MTS分析,发现处以0 3 μM的JA 48小时,即可有效抑制口腔癌细胞OC-2和HSC3的增生,但不影响正常口腔上皮细胞生长。OC-2经JA处理后,其sub-G1的DNA含量增加,并可观察到DNA固缩和梯度断裂的现象,说明JA可使OC-2走向细胞凋亡。除此之外,0 3 μM的JA亦可有效降低OC-2的迁移能力。

Results Nicotinamide at 1-5 mg/ml induced apoptosis of GLC-82 cells in a dose-dependent manner, and group 4 mg/ml and group 5 mg/ml showed increased apoptosis than other groups. Distinct morphologic changes of cell apoptosis such as karyopyknosis and conglomeration were observed; Cell migration became slower as the dosage of Nicotinamide increased;Cells produced an arrested at G1 phase, accompanied by a reduction of cells transiting through S phase.

结果 烟酰胺诱导细胞凋亡程度与用药剂量呈正相关,较高剂量的药物能引起GLC-82肺腺癌细胞呈现边界不清,体积缩小,核固缩,染色质凝集、边缘化,甚至细胞发生破碎,凋亡小体形成;随着烟酰胺浓度的增高GLC-82细胞向划痕区的愈合速度不断减慢;烟酰胺的浓度大于等于3 mg/ml 时,细胞周期主要表现为G1 期比例的上升伴随S期比例的下降。

Result 1 Magnetic nanoparticles, magnetic nanoparticles modified with antisense oligodeoxynucleotide of human telomerase reverse transcriptase induced HL-60 tumor cells to apoptosis, we could see typical morphologic change of apoptosis cells: karyopyknosis, chromation"s condensing and aggregation in nuclear, forming crescent-shaped or annulus structures to lean on edge of cell nucleus"s membrane and posing apoptosis body by Atomic Force Microscope, Fluorescence microscope, transmission electron Microscope 2 There was a significant difference compared with control group(p.01), inhibition ratio had significant positive correlation with medication dosage and time ;during 0.8-8μM dosage amplitude, inhibition ratio accrescenced by dosages increasing. However, the inhibition ratio would decrease when dosage over 8-80μM.

结果 1 磁性纳米粒子、修饰有端粒酶反义寡核苷酸的磁性纳米粒子诱导HL-60细胞发生凋亡,原子力显微镜、光学显微镜、荧光显微镜和透射电镜下均观察到HL-60细胞呈现典型的凋亡细胞的形态变化:细胞核固缩,核内染色质浓缩、凝聚、形成新月形或环状结构紧靠在细胞核膜边缘,并形成凋亡小体。2 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子对HL-60肿瘤细胞的生长和增殖有明显的抑制作用,与对照组相比有显著性差异(p<0.01),在剂量为0.8-8μmol/L范围内,抑制率随剂量的增加而增加,当剂量超过8μmol/L时,抑制率反而下降;3 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子可增强p53基因的表达活性,引起DNA降解损伤,反向调节细胞周期活动,促使细胞从G0期进入G1期,抑制肿瘤细胞的生长。4 修饰有端粒酶反义寡聚脱氧核苷酸的量子点能通过内吞作用进入HL-60肿瘤细胞的细胞核,可以在细胞内进行定位和促进HL-60肿瘤细胞的凋亡。

Methods: A brain concussion was produced in rats using a simple pendulum device model. One control (n=12) and six experimental groups were used(n=12 in each group). The cerebral concussion groups were assigned to sacrifice times ranging from 1 day to 24days after injury.A Morris Water Maze memory paradigm was used to assess learning and memory function. Pycnosis degeneration or necrosis neurons in the locus cerebral cortex, dorsal hippocampus, dentate fornix and brainstem reticular formation were observed and counted after brain concussion. Cholinergic neurons in the basal forebrain and brainstem reticular formation were identified and quantitated using choline acetyltransferase.

应用自制单摆式闭合性机械打击装置建立脑震荡大鼠模型;成年SD大鼠随机分为1个对照组(n=12)和6个脑震荡组(即伤后1、2、4、8、16、24d,n=12);应用Morris水迷宫(Morris Water Maze,MWM)实验评价大鼠的学习记忆功能;用光镜对大鼠的上述脑区的固缩变性坏死细胞进行观察和计数;用免疫组化方法检测基底前脑和脑干网状结构的胆碱乙酰转移酶(cholin acetyltransferase,ChAT)阳性细胞的表达。

That the neurons became swollen at early period, and at later period edematous changes became obvious, and the nuclei of neurons became pyknotic, all displaying specially in CAI region. There were also some microglial cells around the injured neurons.

结果大 鼠的早期行为表现较为兴奋,而后随时间的延长表现为迟钝;神经细胞早期为水肿,后期为水肿明显和细胞器的破坏直至细胞的固缩,这些损伤主要表现在CA1区;同时在损伤的神经元周围常可见小胶质细胞的存在。

Results:①The amount of human colon carcinoma cell line SW480 treated by quercetin decreased. The morphology of partial SW480 cells was shrunk volume, integrated cell membrane, condensed cytoplasm, pyknotic chromatin, nuclear fragmentation. Apoptotic Corpuscles were found by electron microscope.②MTT colorimetric assay showed quercetin inhibited the growth of human colon carcinoma cell line SW480 in a time- and dose-dependent manner when the concentration of quercetin was 30、60、90μmol/L.③Flow cytometry analysis showed the cell cycle of SW480 cell was restricted in G1/S. G0/G1 phase rate increased and S phase rate decreased with increasing concentration of quercetin and time lasting.④ Zymogram analysis assay showed the secretion of matrix metalloproteinases in human colon carcinoma cell line SW480 treated by quercetin decreased. With increasing concentration of quercetin, the secretion of MMP-2 and MMP-9 decreased.⑤Immunohistochemistry method demonstrated the position expression of Cathepsin-D in SW480 cell was suppressed by quercetin in a time- and dose-dependent manner.

研究结果:经槲皮素处理的人结肠癌SW480细胞数量减少,部分细胞体积缩小,细胞膜完整,胞浆浓缩,核染色质固缩,细胞核碎裂,形成凋亡小体;MTT法检测显示当作用浓度为30μmol/L~90μmol/L时,槲皮素对人结肠癌SW480细胞的生长有抑制作用,其抑制作用随着作用浓度的增加和作用时间的延长而增强;流式细胞学发现槲皮素主要作用于人结肠癌SW480细胞周期的G1/S期,大部分细胞被阻断于S期,随药物浓度的升高和作用时间的延长,G0/G1期细胞比例逐渐增加,S期细胞比例逐渐减少;酶谱分析法检测显示不同浓度的槲皮素能够抑制人结肠癌SW480细胞分泌MMP-2及MMP-9,随浓度的升高,MMP-2及MMP-9的分泌量减少;免疫组织化学法显示不同浓度的槲皮素处理人结肠癌SW480细胞后,Cathepsin-D的表达随药物浓度的升高和作用时间的延长而降低。

While the tissue spaces surrounding a few blood vessels wasAl and Fg positive,no Al or Fg positive cells were observed.In antemortem injurygroup,diffuse subarachnoid hemorrhage,cerebral edema,swelling or pyknotic neu-rons could be observed.The axons showed irregular swelling and disconnection at1~3h,marked swelling and disconnection at 6h,and retraction ball at 15h whichwas more remarkable at 24h after injury.The space between myelin sheaths andaxons was increased at 3~6h after injury.Tortuous and wavelike myelin sheathswhich adhered on axons incompletely,or even peeled off could be found from 15hto 24h after injury.Perinuclear lysis of Nissl bodies began at 24h after injury.Thenumber of GFAP positive cells in cerebrum and brain-stem increased significantlyfollowed by decrease,and then increased again,but the time courses of the changesin different areas of brain were not same.Al and Fg positive neural cells,mainlysurrounded blood vessels,with diffuse or peripherally distributed positive matter incytoplasm could be observed at 0.5h after injury.The number of Al or Fg positivecells and the intensity of immunoreaction increased with the time of injury.The areaof SYN positivity in medulla oblongata and pons decreased notably 3~6h afterinjury,then return to normal levels and continued to 24h after injury.

生前损伤组,可见广泛蛛网膜下腔出血,脑组织水肿,神经细胞肿胀,晚期神经元固缩;伤后1~3h见部分神经轴突不规则增粗、断裂,伤后6h断端膨大,伤后15h可见收缩球,至伤后24h更为明显;伤后3~6h可见部分神经髓鞘与轴突之间的间隙增宽,伤后15h髓鞘明显曲折,不完全附着在轴突两侧,甚至剥脱,持续到伤后24h;核周尼氏体减少在伤后24h才开始出现;同一部位的GFAP阳性细胞数目随损伤时间发生改变,先增多(最早在伤后0.5h),达到高峰后减少,其后又有增多趋势,但不同部位的GFAP阳性细胞数目增减的时间过程不尽相同,同时,大脑中的GFAP阳性细胞数目也有改变;伤后0.5h,可在脑干组织中见到Al和Fg阳性神经细胞,主要位于血管周围,阳性物在胞浆中呈弥散性分布,但部分细胞的阳性物仅分布于靠近胞膜的胞浆中而呈环状,随损伤时间延长,阳性细胞数目增多,反应强度增加;伤后3~6h,延髓及桥脑中的SYN阳性物面积减少,其后恢复到正常水平,并持续到伤后24h。

After administration of L-N〓-nitro-Arginine, a specific antagonist of NOS, it was found that the basal and stimulated NO release of blood vessels (aorta, internal carotid, renal and mensenteric artery) was decreased further. The increase of coronary blood flow stimulated by Ach was also reduced further, while the platelet aggregation degree and erythrocyte aggregation index increased significantly. Pathological examination indicated that aortic endothelium was destroyed, the blood vessels in cerebral, cardiac and renal tissues were hypertrophied further and some were almost occluded. The above fin dings resulted in SHR feeded with L-NNA having high incidence of stroke rate and high degree of renal cirrhosis when compared to control SHR. The infarct and hemorrhagic focuci in cerebral tissues, infarct focuci in cardiac tissues, atrophy and hyalinedegeneration of renal glomeruli and degeneration even necrosis of renal tubuli in renal tissues were al so found. The increase ratio of brain and heart to body weight and decrease ratio of kidney to body wei ght further demonstrated that the cerebral, cardiac and renal injuries were aggravated.

在上述结果的基础上进一步给予SHR NOS特异性拮抗剂左旋亚硝基精氨酸后,血管(胸主、颈内、肾及肠系膜动脉)NO基础释放及激动剂刺激释放进一步减少,Ach刺激冠脉流量的增加和胸主动脉内膜NOS活性也进一步减少,血小板聚集程度及红细胞聚集指数显著上升,病理切片显示胸主动脉内皮细胞破坏明显,脑心肾组织内的血管进一步增厚甚至几乎阻塞,导致饲L-NNA的SHR缺血性及出血性脑卒中发生率显著高于对照SHR,脑体重比显著升高;心肌内出现明显的缺血梗塞疤痕灶,心体重比显著升高;肾小球明显萎缩、玻璃样变性,肾小管也变性甚至坏死,使肾硬化程度显著增加呈颗粒性固缩肾,肾体重比也显著减少。

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