白藜芦醇诱导人乳腺癌MDA-MB231细胞凋亡及自噬

目的探讨白藜芦醇(resveratrol,Res)诱导人乳腺癌MDA-MB231细胞凋亡及自噬蛋白表达的作用与机制。方法以人乳腺癌MDA-MB231细胞为研究对象,MTT法检测Res(0、1、5、10、20、50、100、200 mg·L~(-1))抑制MDA-MB231细胞增殖情况; Res(0、5、20、60 mg·L~(-1))处理MDA-MB231细胞,划痕实验检测细胞迁移能力,Annexin V/PI双染法检测细胞早期凋亡率,免疫荧光法检测细胞LC3Ⅰ/Ⅱ的表达;Res(0、20、60 mg·L~(-1))处理MDA-MB231细胞,蛋白免疫印迹法检测细胞中自噬相关蛋白p62及Beclin-1的表达。结果Res在体外可明显抑制MDA-MB231细胞增殖,且呈浓度依赖关系。随着Res浓度的增高,乳腺癌细胞迁移能力较对照组分别下降了(8. 7±1. 1)%、(16. 5±2. 9)%和(32. 2±3. 6)%。流式细胞仪检测结果表明,早期凋亡细胞数增加;激光共聚焦检查显示,细胞质中LC3Ⅰ/Ⅱ绿色荧光增加;蛋白免疫印迹法检测出MDA-MB231细胞p62蛋白表达下降,Beclin-1蛋白表达升高。结论 Res可抑制人乳腺癌MDA-MB231细胞增殖,其机制可能与Res诱导细胞凋亡和自噬有关。     

白花蛇舌草总黄酮体外对人肝癌细胞HepG-2的作用

目的:观察白花蛇舌草总黄酮体外对HepG-2肿瘤细胞增殖的抑制作用。方法:以HepG-2细胞为实验细胞株,观察白花蛇舌草中的总黄酮对HepG-2肿瘤细胞形态的影响;用流式细胞仪检测细胞的凋亡。结果:白花蛇舌草总黄酮对HepG-2肿瘤细胞形态有影响,抑制其生长并诱导HepG-2肿瘤细胞的调亡。结论:白花蛇舌草中的总黄酮通过诱导凋亡对人肝癌细胞HepG-2有抑制增殖作用。     

沙枣提取物对某些肿瘤细胞株及小鼠实体瘤和腹水瘤的抑制作用

目的探讨沙枣提取物对4种肿瘤细胞株及小鼠实体瘤和腹水瘤的抑制作用及机制。方法应用MTT法检测了提取物对4个肿瘤细胞株生长的抑制作用,检测提取物对实体瘤肿瘤生长的抑制率和腹水瘤小鼠的生命延长率,探讨提取物对肿瘤细胞凋亡的影响及机制。结果体外实验中观察到高中低剂量组具有明显抑制4种肿瘤细胞生长作用(P<0.01)。体内实验观察到沙枣提取物高中低剂量均有抑制S180实体瘤小鼠肿瘤生长、延长S180腹水瘤小鼠的生存时间作用。形态学观察发现提取物诱导了4种肿瘤细胞的凋亡,提取物诱导4种肿瘤细胞早期凋亡率在14.1%～24.5%之间,明显高于未处理肿瘤细胞(P<0.05)。提取物处理T24细胞后,bax、caspase-3蛋白表达明显增强,bcl-2表达降低。结论在本试验条件下,沙枣提取物有抑制肿瘤生长作用,其机制可能与下调bcl-2表达和上调bax的表达,高表达caspase-3诱导肿瘤细胞凋亡有关。     

一种新的吲哚咔唑类化合物(ZWM233)的体外抗肿瘤作用及机制探讨

目的探讨ZWM233体外对多种肿瘤细胞株的增殖抑制及对K562细胞的凋亡诱导作用。方法采用MTT法检测ZWM233对K562、HL-60、A549、HeLa及HCT-116等10种肿瘤细胞株的增殖抑制作用;流式细胞仪检测对K562细胞周期的影响;AnnexinV/PI双染法考察对K562细胞的凋亡诱导作用;Western blot检测K562细胞中凋亡相关蛋白bax、bcl-2、Caspase-3、Caspase-9及PARP的变化,同时检测蛋白激酶C(PKC)各亚型、GSK-3β、ERK1/2、JNK及其磷酸化水平的变化。结果 ZWM233体外能有效抑制多种肿瘤细胞株的增殖,其中对K562细胞作用明显,IC50为2.81μmol.L-1;流式细胞仪检测K562细胞被明显阻滞在G2/M期,并诱导其凋亡;Western blot结果显示ZWM233作用K562细胞24 h后,可明显下调抗凋亡蛋白bcl-2的表达,引起Caspase-9、Caspase-3及下游底物PARP的剪切,诱导K562细胞凋亡。Western blot检测结果进一步显示药物作用24 h后PKCδ的表达有所降低,其下游分子p-GSK-3β及p-ERK的表达水平降低,而p-JNK蛋白的表达水平升高。结论ZWM233体外能有效抑制K562细胞生长,并通过激活线粒体途径诱导其凋亡,其机制可能是通过下调PKCδ,进一步抑制p-GSK-3β及p-ERK1/2的表达来发挥凋亡诱导作用。     

ERK抑制剂U0126通过下调cyclin D1与survivin蛋白表达抑制乳腺癌细胞增殖

目的研究ERK通路抑制剂U0126对乳腺癌细胞增殖的抑制作用,并探讨其调控机制。方法培养人乳腺癌细胞株MCF-7、MDA-MB231,将细胞分组干预,MTT法检测各组细胞增殖情况;流式细胞仪检测各组细胞周期及细胞凋亡; Western blot检测细胞中p-ERK/ERK、cyclin D1、survivin及cleaved caspase-3蛋白表达。结果MTT结果显示,U0126干预24、48 h后,MCF-7、MDA-MB231细胞抑制率明显增加(P <0. 01); MCF-7、MDA-MB231细胞经U0126干预24 h后,检测细胞周期及细胞凋亡,G_0/G_1期细胞比例较对照组明显增加,S及G_2期细胞比例下降(P <0. 05);而干预组细胞凋亡率较未干预组明显增多(P <0. 01); U0126处理人乳腺癌细胞2 h后,可阻断ERK的磷酸化,减少cyclin D1的蛋白表达,抑制survivin而上调cleaved caspase-3的表达,与对照组相比差异有显著性(P <0. 01)。结论 U0126通过阻断MCF-7、MDA-MB231细胞中ERK信号通路,调控cyclin D1,将细胞周期阻断在G_0/G_1期,抑制survivin而增加cleaved caspase-3表达,增加细胞凋亡,继而使乳腺癌细胞MDA-MB231、MCF-7的增殖受到抑制。     

啤酒花中蛇麻酮体外诱导人胃癌SGC-7901细胞凋亡作用研究

目的探讨啤酒花(Humulus LupulusL.)中成分蛇麻酮(Lupulone,LP)体外对人胃癌细胞SGC-7901生长抑制及诱导凋亡作用的研究。方法体外培养人胃癌细胞株SGC-7901,MTT法检测不同浓度蛇麻酮对SGC-7901细胞体外生长的影响;流式细胞仪分析蛇麻酮对SGC-7901细胞的凋亡率的影响。结果蛇麻酮对SGC-7901的生长具有较强的抑制作用,IC50为0.73μg/mL;0、0.2、0.8、3.2μg/mL剂量的蛇麻酮作用SGC-7901细胞48h细胞凋亡率分别为0.1%、7.1%、18.0%、49.3%。结论蛇麻酮对SGC-7901具有较强的抗肿瘤活性,其机制可能与其可诱导肿瘤细胞凋亡有关。     

HSPG和无HS-GAGs链HSPG体外抗乳腺癌的研究

目的：比较硫酸乙酰肝素蛋白聚糖（简称HSPG）和去硫酸乙酰肝素链（简称HS-GAGs链即HS链）的HSPG的体外抗人乳腺癌细胞MDA-MB-231的抗增殖和凋亡诱导作用。方法：对HSPG进行提取、纯化,硫酸乙酰肝素酶1水解HSPG的HS链;光学显微镜法观察两组分引起的细胞形态学的变化;MTT比色法检测两组分对肿瘤细胞生长曲线的影响;流式细胞术结合Annexin V-FITC及PI双染标记法观察两组分对肿瘤细胞凋亡的影响。结果：HSPG可致MDA-MB-231细胞脱落悬浮;MTT法检测结果显示HSPG对MDA-MB-231细胞增殖的抑制作用随浓度的增加而增强,而去HS链HSPG只有在高浓度时才呈现出抑制作用。HSPG能诱导MDA-MB-231的早期凋亡,而去HS链HSPG在高浓度（4 mg/mL）时对MDA-MB-231早期凋亡有诱导作用。结论：HSPG能诱导MDA-MB-231的早期凋亡和抑制细胞生长增殖。作用机制可能与HSPG的硫酸乙酰肝素链有关。     

红毛五加提取物诱导卵巢癌细胞系OVCAR3凋亡

目的研究红毛五加提取物的抗肿瘤活性。方法分别采用MTT法、形态学、流式细胞术方法检测纯氯仿洗脱部分对肿瘤细胞OVCAR3的生长抑制及促凋亡。结果 MTT法可见氯仿洗脱部分对体外培养的卵巢上皮癌细胞OVCAR3的生长有抑制作用,高、低剂量组抑制率分别为20.6%、11.4%;形态学以及流式细胞术都可见氯仿洗脱部分可以促进OVCAR3凋亡。结论红毛五加纯氯仿洗脱部分对肿瘤细胞株OVCAR3细胞有促凋亡的作用。     

鱼腥草黄酮提取物对肿瘤细胞的抑制作用

目的研究鱼腥草黄酮类提取物在体外对人白血病细胞HL60、小鼠黑色素瘤细胞株B16BL6增殖抑制和凋亡的影响.方法(1)采用纯化水回流提取,大孔树脂分离的方法提取鱼腥草中黄酮类化合物;(2)应用MTT法检测黄酮类化合物对HL60和B16BL6肿瘤细胞株的细胞抑制率;(3)采用FCM法检测其对肿瘤细胞凋亡的影响.结果(1)鱼腥草黄酮提取物有抑制HL60和B16BL6细胞生长的作用,50%增值抑制浓度值分别为0.410,0.122 g·L-1,(2)鱼腥草黄酮提取物能诱导HL60和B16BL6细胞的凋亡.结论鱼腥草黄酮提取物能抑制HL60和B16BL6肿瘤细胞生长,具诱导凋亡的作用,为鱼腥草的抗肿瘤效应提供实验依据.     

低分子量肝素对骨肉瘤细胞生长与凋亡的影响研究

目的 研究低分子量肝素（LMWH）体外抗肿瘤生长及诱导肿瘤细胞凋亡的作用,初步探讨低分子量肝素抗肿瘤作用机制,为该药的抗肿瘤作用开发和临床应用提供试验依据.方法在体外培养的骨肉瘤系（MG-63）细胞培养基中加入不同浓度的LMWH,对照组加完全培养基,用MTT法检测不同浓度LMWH在干预不同时间后对骨肉瘤细胞生长的影响,流式细胞仪检测不同浓度干预组细胞凋亡率的变化.结果LMWH可抑制骨肉瘤细胞系MG-63生长,并具有时间剂量依赖性,在LMWH浓度为200 U&#8226;mL^-1时,对细胞生长有明显的抑制作用. 流式细胞仪检测显示,随着LMWH剂量的增加,MG-63细胞凋亡率明显增加. 结论LMWH在体外具有抗肿瘤活性,其作用可能通过抑制细胞增殖和促进肿瘤细胞凋亡而实现.     

The effect of quinidine on the analgesic effect of codeine

Summary We have studied the hypoalgesic effect of codeine (100 mg) after blocking the hepatic O-demethylation of codeine to morphine via the sparteine oxygenase (CYP2D6) by quinidine (200 mg). The study was performed in 16 extensive metabolizers of sparteine, using a double-blind, randomized, four-way, cross-over design. The treatments given at 3 h intervals during the four sessions were placebo/placebo, quinidine/placebo, placebo/codeine, and quinidine/codeine. We measured pin-prick pain and pain tolerance thresholds to high energy argon laser stimuli before and 1, 2, and 3 h after codeine or placebo.After codeine and placebo, the peak plasma concentration of morphine was 6–62 (median 18) nmol·.l^–1. When quinidine pre-treatment was given, no morphine could be detected (<4 nmol·l^–1) after codeine. The pin-prick pain thresholds were significantly increased after placebo/codeine, but not after quinidine/codeine compared with placebo/placebo. Both placebo/codeine and quinidine/codeine increased pain tolerance thresholds significantly. Quinidine/codeine and quinidine/placebo did not differ significantly for either pin-prick or tolerance pain thresholds.These results are compatible with local CYP2D6 mediated formation of morphine in the brain, not being blocked by quinidine. Alternatively, a hypoalgesic effect of quinidine might have confounded the results.     

罗森塔尔效应在急诊带教中的应用效果分析

目的分析罗森塔尔效应在急诊带教中的应用效果。方法选取我院2015年7~2016年4月期间的急诊实习学生共61例,按照双盲法分为观察组(n=31)和参照组(n=30),参照组实习学生应用常规带教教学方式,观察组实习学生应用罗森塔尔效应带教教学方式,对比分析两组实习学生的考核成绩和护理满意评分。结果在考核成绩对比中,经一段时间学习后,观察组实习学生无论是理论成绩还是实践成绩均优于参照组,差异有统计学意义(P<0.05);在护理满意评分对比中,观察组实习学生在学习主动性、学习能力、专业技能、自律性及总分等护理满意评分中均高于参照组,差异有统计学意义(P<0.05)。结论急诊带教中应用罗森塔尔效应的效果显著,能够提高学生学习自主性、专业知识掌握程度和专业技能,具有较高的实践应用价值,有利于学生更快更好的适应急诊病房环境。     

罗格列酮的胰岛素增敏作用和胰岛素抵抗改善作用

目的　观察罗格列酮的胰岛素增敏作用和胰岛素抵抗改善作用。方法　大鼠尾静脉注射链佐菌素 (5 0mg·kg-1)制备IDDM模型 ,观察单用罗格列酮、单用小剂量胰岛素 (皮下注射 2 5U·鼠 -1)、罗格列酮并用小剂量胰岛素的降糖作用 ;大鼠尾静脉注射卡介苗 (10mg·鼠 -1)制备免疫性胰岛素抵抗模型 ,观察罗格列酮对模型的葡萄糖输注速率的回升作用。结果　罗格列酮 1,3 ,10 μmol·kg-13个剂量组连续灌胃给药 8d ,均未能降低正常大鼠血糖 ,也未能降低链佐菌素所致糖尿病大鼠血糖 ;但在并用小剂量胰岛素条件下 ,罗格列酮 3 ,10 μmol·kg-1连续灌胃给药 8d ,能降低链佐菌素所致糖尿病大鼠血糖 ;罗格列酮 10 μmol·kg-1连续灌胃给药 8d后 ,用正糖钳技术检测 ,免疫性胰岛素抵抗模型亚极高和极高胰岛素状态下的葡萄糖输注速率均得到回升。结论　罗格列酮具有胰岛素增敏作用和胰岛素抵抗改善作用     

The effect of carbon disulphide on the stereotypic effect of dopamine agonists

Exposure to CS₂ increased the intensityof apomorphine-induced stereotypy in male rats without increasing the reaction time. With amphetamine, an indirect agonist of dopamine, exposure to CS₂ had a more intensive effect and significantly prolonged the length of reaction.     

Pressor effect of indolethylamines

6-Hydroxytryptamine, 7-hydroxytryptamine and 5,7-dihydroxytryptamine increase blood pressure when given intravenously to pithed rats and effect not due to stimulation of 5-hydroxytryptamine receptors. 6-Hydroxytryptamine and 7-hydroxytryptamine interact with α-adrenoceptors of the vascular bed. An effect of 5,7-dihydroxytryptamine on α-adrenoceptors was not established.     

Vasorelaxant effect of harman

The in vivo cardiovascular effect and in vitro vasorelaxant effect of harman, one of harmala alkaloids isolated from Peganum harmala, were examined in this study. Harman (1-10 mg/kg, i.v.) dose-dependently produced transient hypotension and long-lasting bradycardia in pentobarbital-anesthetized rats, which were attenuated by N(G)-nitro-L-arginine pretreatment. In isolated rat endothelium-intact thoracic aortic rings, harman concentration dependently relaxed phenylepherine-induced contractions with an IC(50) value around 9 microM. This vasorelaxant effect was attenuated by endothelium removal or N(omega)-nitro-L-arginine methyl ester pretreatment, but not by tetraethylammonium or indomethacin pretreatment. In cultured rat aortic endothelial cells, harman (1-100 microM) concentration dependently increased nitric oxide (NO) release, which was dependent on the presence of external Ca(2+). Harman pretreatment (3-30 microM) also concentration dependently inhibited the contractions induced by phenylephrine, 5-hydroxytryptamine (5-HT), and KCl in endothelium-denuded aortic rings in a non-competitive manner. In addition, harman pretreatment reduced both phasic and tonic phases of phenylephrine-induced contractions. Receptor binding assays further indicated that harman (K(i) values around 5-141 microM) interacted with the cardiac alpha(1)-adrenoceptors, brain 5-HT(2) receptors, and cardiac 1, 4-dihydropyridine binding site of L-type Ca(2+) channels. Therefore, the present results suggested that the vasorelaxant effect of harman was attributed to its actions on the endothelial cells to release NO and on the vascular smooth muscles to inhibit the contractions induced by the activation of receptor-linked and voltage-dependent Ca(2+) channels. The vasorelaxant effect may be involved in the hypotensive effect of harman.     

Post-antibacterial effect of thymol

The antibacterial activity of thymol has been well established and reported in the scientific literature. Continued suppression of bacterial growth following limited exposure to antimicrobial compounds at different concentrations greater than or equal to the minimum inhibitory concentration level (MIC) and at concentrations less than the MIC can be used as an indicator of biological activity, and are respectively referred to as a post-antibacterial effect (PAE) and a post-antibiotic sub-MIC effect (PA-SME). In this study, the PAE and the PA-SME of thymol against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus were investigated. A spectrophotometric method was used to determine the PAE and the PA-SME of thymol against the selected test strains. Thymol exhibited a considerable PAE and PA-SME at MIC and sub-MIC concentrations against test strains. The greatest duration of both the PAE and the PA-SME was observed for thymol against E. coli and P. aeruginosa. The PAE and PA-SME times for E. coli were 12 and 8?h, respectively, and for P. aeruginosa were 11 and 7.5?h, respectively. The duration of the PAE and PA-SME observed for S. aureus and B. cereus was shorter than for Gram-negative strains.     

黄谷素抗心律失常作用及其机制

黄谷素能明显对抗大鼠乌头碱、冠脉结扎诱发的心律失常；对乌头碱引起犬的心律失常有明显治疗作用。在膜片钳实验中发现，黄谷素能显著抑制豚鼠左房单细胞延迟外向钾电流和尾电流，这一作用具有浓度依赖性。黄谷素１０μｍｏｌ·Ｌ－１使ＩＫ从给药前的１１６０ｐＡ减少到８１０ｐＡ，使尾电流从给药前的３３１ｐＡ减少到１９６ｐＡ，此作用在膜电位０～６０ｍＶ区间作用较强。黄谷素抑制延迟外向钾电流和尾电流是其抗心律失常作用机制。     

α-菠菜甾醇的抗炎作用

α-菠菜甾醇对动物角叉菜胶和热烫性足肿胀、巴豆油气囊肿肉芽组织增生均有明显抑制作用,其抗炎作用机理可能抑制PGE₂、缓激肽的合成或释放。明显抑制PGE₂、缓激肽、组胺、5-HT等炎症介质的致炎作用,抑制白细胞游走。并证明其抗炎作用不依赖于肾上腺垂体系统,但对肾上腺皮质功能也有明显的促进作用。     

肝素的平喘作用

目的：探讨肝素的平喘作用。方法：采用豚鼠整体动物引喘法和兔改良肺溢流法观察肝素的平喘作用；采用Ｇｒｅｅｎ法观察肝素对豚鼠血清一氧化氮（ＮＯ）含量的影响；采用二甲苯诱发小鼠耳肿胀法以及大鼠松节油气囊肉芽肿法观察肝素对炎症的影响；采用小鼠断尾法和豚鼠玻管法观察肝素对出凝血时间的影响。结果：肝素气雾给药，５２０．８３、１０４１．６６Ｕ·ｋｇ－１可使豚鼠引喘潜伏期延长；５２０．８３Ｕ·ｋｇ－１可使兔肺溢流量减少；１０４１．６６Ｕ·ｋｇ－１可使豚鼠血清ＮＯ含量增加。但等剂量的肝素对动物的急、慢性炎症及出凝血时间无明显影响。结论：小剂量肝素气雾给药有明显的气道扩张及平喘作用；此剂量肝素的应用可避免出血倾向等不良反应的发生；其平喘１９９７－０１－３１收稿，１９９７－０３－０２修回作者简介：明亮，女，４９岁，副教授作用可能与抗炎作用无直接关系；ＮＯ含量升高与其平喘作用有何联系，尚待研究。