层黏连蛋白受体反义核酸对卵巢癌细胞侵袭能力的影响

目的:研究67 kD层黏连蛋白受体(67kDLN-R)反义寡核苷酸(antisense oligonucleotide,ASODN)对卵巢癌细胞HRA体外侵袭转移能力的影响.方法:采用流式细胞仪、RT-PCR及Transwell小室的方法检测反义核酸对67 kD LN-R基因和蛋白表达影响,及转染前后细胞体外侵袭转移能力变化.结果:67 kD LN-R ASODN可从蛋白质和mRNA水平下调67kDLN-R的表达,且下调作用呈剂量依赖性.与正义组及对照组相比差异有显著性(P＜0.05).体外侵袭实验证实转染后HRA穿透人工基底膜的能力显著降低并呈现剂量依赖性.结论:67kDLN-R ASODN可降卵巢癌细胞体外侵袭转移能力,有望为卵巢癌治疗提供新方向.     

RT-PCR检测乳腺癌患者外周血表皮生长因子受体和细胞角质蛋白-19mRNA的表达

目的　探讨逆转录聚合酶链式反应(RT-PCR )方法检测乳腺癌患者外周血表皮生长因子受体(EGFR)mRNA作为乳腺癌循环肿瘤细胞标志的可能性及意义。方法　以转录聚合酶链式反应(RT-PCR)方法,分别对40例乳腺癌患者和30例健康人静脉血进行EGFRmRNA检测,同时与目前研究较多的循环上皮细胞标志:细胞角质蛋白CK-19的mRNA做比较。结果　在30例健康人外周静脉血中没有发现EGFRmR NA的表达( 0% ),但有8例CK-19阳性( 26. 7% )。而40例乳腺癌患者检测出13例EGFRmRNA阳性(32. 5% ),与正常人相比P<0. 01,两组间存在显著性差异;CK-19的mRNA阳性例数为14 (35. 0% ),与正常人相比P>0. 05,两组间无统计学差异。通过免疫组化检测到25例( 62. 5% )的肿瘤组织中EGFR阳性,所有外周血中EGFRmRNA阳性的病例,相应的原发肿瘤病灶中EGFR蛋白均为阳性,组织中EGFR蛋白高表达的患者外周血EGFRmRNA检出率也高(P<0. 01)。26例初治患者中5例EGFRmRNA阳性(19. 2% ),14例复治患者中8例EGFRmRNA阳性(57. 1% ),两组相比P<0. 05,存在显著性差异。6例曾使用过紫杉醇类药物挽救化疗的转移性患者中1例EGFRmRNA阳性(16. 7% ), 8例未使用过紫杉醇类药物挽救化疗的转移性患者中7例EGFRmRNA阳性(87. 5% ),两组相比P<0. 05,存在显著性差异。结论　CK-19的     

人UrotensinⅡ在心血管系统中的生物学效应

Urotensin II最早是从鱼尾部下垂体中分离出的神经环肽,随后在一种欧洲绿蛙Rana ridibunda的脑中鉴别出来和从人体基因组中克隆出来。人Urotensin II(human urotensin II,hUII)有11个氨基酸残基组成,主要分布于心血管和神经系统。作为UT受体的一种内源性配体,hUII有广泛的生物学效应,如收缩和舒张血管,促进心肌细胞增殖,调节心功能,并在冠心病、心功能衰竭等某些心血管疾病的病理过程中起着重要的作用。本文就hUII在心血管系统方面的生物学效应作一简要的综述。     

坎地沙坦,PD123319及金雀异黄素对大鼠血管平滑肌细胞迁移影响

目的:探讨血管紧张肽Ⅱ(AngⅡ)不同受体亚型(AT1,AT2)在血管平滑肌细胞(VSMC)迁移中的作用。方法:以体外培养VSMC为基础,采用改良Boyden小室,对不同浓度AT1拮抗剂坎地沙坦(CV)、AT2拮抗剂PD123319(PD)和酪氨酸激酶(PTKs)抑制剂金雀异黄素作用下AngII诱导产生的VSMC跨膜迁移细胞数进行评价。分对照组、AngII组、AngII+10-10～10-5mol·L-1CV组、AngⅡ+10-9～10-6mol·L-1PD组、AngⅡ+CV+PD组、AngⅡ+金雀异黄素组。结果:与对照组相比,AngⅡ组跨膜迁移细胞数明显增加,P<0.01。坎地沙坦在10-10～10-5mol·L-1的浓度范围内,呈剂量依赖性抑制由AngⅡ介导的VSMC迁移(r=0.95,P<0.05)。PD123319对此无明显的增强或抑制作用。金雀异黄素组VSMC跨膜迁移细胞数低于AngⅡ组,高于对照组。结论:AngⅡ影响VSMC迁移能力的生物学效应由AT1介导;AT2在VSMC迁移过程中不起作用或者不起主要作用。酪氨酸激酶的激活也参与了AngⅡ诱导的VSMC迁移的信号转导过程。     

氨甲酰胆碱和毛果芸香碱对血管内皮细胞M受体基因表达的调节作用

目的：比较氨甲酰胆碱和毛果芸香碱对血管内皮细胞M受体基因表达调节作用的异同点。方法：氨甲酰胆碱和毛果芸香碱分别孵育培养的牛主动脉内皮细胞10h后，提取细胞总RNA，RT-PCR方法测定M受体5个亚型mRNA的表达。结果：M_1～M_5受体mRNA在牛主动脉内皮细胞上均有表达，氨甲酰胆碱和毛果芸香碱孵育后表达量均增加，但二者之间无差异。结论：氨甲酰胆碱和毛果芸香碱对M受体5个亚型的基因表达均发生了相同的影响，两者对M受体可能的作用尚不能解释其对血管张力的影响。     

心肌细胞膜固定相色谱研究中药川芎提取液与受体的作用

目的 :研究中药川芎提取液与心肌细胞膜受体的作用。方法 :将家兔心肌细胞膜固定在硅胶载体上制成色谱固定相 ,通过川芎水提液和醇提液与心肌细胞膜上已知受体的部分激动剂和拮抗剂在心肌细胞膜色谱柱上保留特性的对比 ,以及流动相中加入心肌细胞膜上已知受体激动剂或拮抗剂时由于竞争结合而产生的对提取液色谱保留特性的影响 ,研究川芎提取液与心肌细胞膜受体的作用。结果 :水提液和醇提液在心肌细胞膜色谱柱上均有保留行为 ;水提液保留特性受α受体阻断剂竞争结合的影响 ,而不受 β₁受体激动剂的影响。结论 :川芎水提液中可能含有与α受体作用的有效成分、不含有与 β₁受体作用的有效成分 ,醇提液中亦含有与心肌细胞膜受体作用的成分。将细胞膜固定在色谱填料上 ,能够进行模仿药物在体内与受体作用的有关过程 ,不仅可以用于药物筛选 ,还可以用于揭示其所作用的受体类型。     

冠心病体质表型和低密度脂蛋白受体基因Avall 位点多态性关系的研究

目的:探讨CHD(冠心病)体质类型和LDL-R(低密度脂蛋白受体)第13外显子AvaⅡ位点基因多态性的关系.方法:选择符合条件的冠心病中医痰证、瘀证、痰瘀互阻证和非痰非瘀证(其他证型)患者200例,判别患者的体质类型.另选100例健康对照,常规检测所有样品的血脂水平.提取全血DNA,经PCR扩增获取目的片段,用AvaⅡ酶切检测LDL-R基因型,采用SAS软件进行相关性分析.结果:患者的-/ 基因型显著多于对照.TC(总胆固醇)、TG(总甘油三酯)和LDL-C(低密度脂蛋白胆固醇)水平在-/-、-/ 、 / 基因型患者中存在渐升的趋势, / 基因型的TC水平和动脉硬化指数AI显著高于-/-基因型.患者的体质类型以瘀血质、痰湿质和气虚质多见,其中痰湿质和 等位基因,瘀血质与 / 基因型密切相关.结论:瘀血质、痰湿质、气虚质是冠心病的主要体质表型.LDL-R第13外显子AvaⅡ位点 等位基因的携带患者多出现在痰湿质和瘀血质中,纯合子患者的TC和AI升高.这为痰瘀互结的中医病机研究提供了初步证据.     

Phospholipid composition and levels are altered in Down syndrome brain

Individuals chronically exposed to low levels of organophosphate insecticides may present with subtle impairments in cognition. In addition, low level diisopropylflurophosphate (DFP) exposure (0.25 mg/kg per day for 2 weeks) in rats resulted in protracted working memory impairment [29]. The current studies attempt to show a temporal relationship between the DFP-induced impairment in performance of a spatial memory task and the protracted decrease in the expression of cholinergic receptors and acetylcholinesterase in specific brain regions. Cholinergic receptors labeled with the ligands [(3)H]epibatidine and [(3)H]AFDX-384 were affected to a much greater extent and for a longer period of time than were both acetylcholinesterase activities and cholinergic receptors labeled with [(3)H]QNB. Pre-testing administration of nicotine was shown to completely reverse this DFP-induced impairment in memory-related task performance. Additionally, prophylaxis with pyridostigmine bromide (PB) caused DFP-treated animals to exhibit near normal levels of memory-related task performance. These results are consistent with the development of a protracted phase of learning impairment to sub-acute DFP exposure, which may involve the loss of hippocampal nicotinic receptors, and may be prevented or reversed by PB or nicotine, respectively.     

Identification of the opioid receptors involved in passive-avoidance learning in the day-old chick during the second wave of neuronal activity

Long-term memory formation for passive-avoidance learning in the day-old chick is known to have two distinct time windows of protein synthesis (F.M. Freeman, S.P.R. Rose, A.B. Scholey, 1995. Two time windows of anisomycin-induced amnesia for passive-avoidance training in the day-old chick. Neurobiol. Learn. Mem. 63, 291-295). The lobus parolfactorius (LPO) is thought to be an important site for the second wave of protein synthesis which occurs 4-5 h after training. Birds received bilateral intracranial injections of agonists and antagonists for the mu-, delta-, kappa-opioid receptors and the opioid receptor-like (ORL(1)) receptor directly into the LPO at 5 h post-training and were tested for recall 24 h later. Also, 100 microM beta-funaltrexamine (beta-FAN), a mu-opioid receptor antagonist, significantly impaired memory formation (P<0.01). The delta-opioid receptor was also involved in memory formation at this time-point since antagonism of this receptor by 1 mM ICI-174,864 caused amnesia (P<0.01) which was reversed by the agonist, DPLPE. The kappa-opioid receptor appeared not to be involved during the second phase of neuronal activity since neither stimulation by dynorphin nor inhibition by nor-BIN caused amnesia for the task. The ORL(1) receptor agonist orphanin FQ also had no effect suggesting that this receptor was not involved at this 5-h time-point. Cytosolic and mitochondrial protein synthesis has been shown to be important in passive-avoidance learning in the day-old chick. Both chloramphenicol (CAP) and anisomycin (ANI), inhibitors of mitochondrial and cytosolic protein synthesis, respectively, caused disruption when injected 5 h post-training into the LPO (P<0.05). Endomorphin-2 (Endo-2), a mu-opioid receptor agonist, reversed both the ANI- and CAP-sensitivity. However, DPLPE, a delta-opioid receptor agonist, only reversed the effect due to CAP. Possible mechanisms for these effects are discussed.