曲古霉素A诱导人膀胱癌细胞周期阻滞和凋亡的研究

目的:探讨组蛋白去乙酰化酶抑制剂曲古霉素A(TSA)对人膀胱癌T24细胞周期和凋亡的影响。方法:以不同剂量TSA(0.1μM,0.3μM和1μM)处理T24细胞。采用MTT法检测细胞存活率,AnnexinV-PI染色检测细胞凋亡,流式细胞仪检测caspase-3活性,Western blot法检测P21蛋白表达。结果:TSA剂量依赖性降低膀胱癌细胞存活率,促进细胞凋亡,表现为AnnexinV阳性细胞明显增多,同时活化的caspase-3水平增高。TSA还可通过诱导膀胱癌细胞周期阻滞于G2/M期抑制细胞生长,且呈剂量依赖性。结论:TSA通过促进caspase-3激活诱导膀胱癌细胞凋亡,同时诱导细胞阻滞于G2/M期。     

姜黄素类似物EF24诱导A549细胞自噬及凋亡关系的研究

从细胞自噬及凋亡关系角度探讨姜黄素类似物EF24对人肺腺癌细胞（A549）的杀伤机理。选用不同浓度的EF24对体外培养的A549处理,采用MTT方法检查细胞存活率,吖啶橙染色观察细胞形态,蛋白质免疫印迹（Western blot）方法检测与细胞自噬及凋亡相关蛋白的表达及对AMPK-mTOR-S6K信号通路的影响。结果显示,EF24作用24 h的IC50为8.5μmol/L,对A549细胞生长抑制作用优于姜黄素,而接近顺铂。自噬及凋亡蛋白检测显示,在4μmol/L、8μmol/L时A549细胞以自噬为主,在16μmol/L时以凋亡为主;加入100 nmol/L自噬抑制剂渥曼青霉素（wortmannin）后,细胞存活率同比升高。同时还发现,随着EF24浓度的增加,细胞内AMPK-Thr172磷酸化水平上升,mTOR-Ser2481、S6K-Thr389磷酸化水平的下调。由此可见,EF24可通过AMPK的激活下调mTOR-S6K途径抑制细胞生长,在EF24浓度4~8μmol/L范围内,自噬对凋亡起到促进作用。     

紫草素促进肺癌A549细胞凋亡

目的:探讨紫草素对A549人肺癌细胞凋亡的影响和可能的作用机制。方法:采用不同浓度的紫草素对体外培养的A549人肺癌细胞进行干预,CCK-8法和流式细胞术分别检测紫草素对A549细胞增殖和凋亡的影响,Western blot观察凋亡相关蛋白(Bcl-2和Bax)表达水平的变化,激光共聚焦显微镜检测紫草素处理12 h并用JC-1染色的A549细胞线粒体膜电位改变。结果:CCK-8分析显示,0.5μM、1μM、2μM、4μM和6μM实验组A549细胞相对存活率分别为(83.71±1.02)%、(57.47±2.78)%、(27.39±1.96)%、(16.96±1.47)%和(14.72±1.93)%,与对照组相比实验组A549细胞相对存活率明显降低;流式细胞术结果表明,1μM、2μM、4μM实验组A549细胞的凋亡率分别为(13.80±1.76)%、(40.90±3.48)%和(78.80±2.52)%,与对照组相比紫草素呈剂量依赖型促进A549细胞凋亡;Western blot结果证实,紫草素能降低A549细胞中Bcl-2蛋白的表达量,而升高Bax蛋白的水平;激光共聚焦显微镜扫描结果显示紫草素能降低A549细胞的线粒体膜电位,呈剂量依赖型。结论:紫草素能显著促进A549细胞凋亡,其机制可能与下调抗凋亡蛋白Bcl-2的表达和上调促凋亡蛋白Bax的表达有关。     

TSA对非小细胞肺癌A549细胞放疗增敏性的研究

目的:探讨组蛋白去乙酰化酶抑制剂(HDIs)trichostatinA(TSA)对非小细胞肺癌(NSCLC)A549细胞放疗敏感性的影响.方法:以5Gy γ-射线照射细胞,或同时以TSA处理细胞.采用MTT法检测细胞存活率,Armexin V-PI染色检测细胞凋亡,流式细胞仪检测caspase-3活性.结果:SGy γ-射线可轻度降低细胞存活率,仅有少量细胞发生凋亡,但同时以TSA和γ-射线处理细胞,细胞存活率明显下降,凋亡细胞显著增多,且明显提高活化的caspase-3水平.结论:TSA通过促进caspase-3激活增强A549细胞对γ-射线的敏感性.     

凋亡诱导因子在姜黄素诱导的大鼠肺成纤维细胞凋亡中的作用

目的:研究姜黄素对肺纤维化大鼠肺成纤维细胞增殖、凋亡的影响,探讨凋亡诱导因子(AIF)在肺成纤维细胞凋亡中的作用.方法:将体外培养的肺纤维化大鼠成纤维细胞,分别于不同浓度的姜黄素(5、10、20、40μM)和caspase-3抑制剂Z-DEVD-fmk(20μM)孵育,观测细胞生长状态变化.MTT检测成纤维细胞增殖抑制率;流式细胞仪检测细胞凋亡率;Western-Blot测定凋亡诱导因子(AIF)蛋白表达及核转位结果:流式细胞术检测细胞凋亡,5～40μM姜黄素处理12 h,其凋亡率呈浓度依赖,对照组相比,差异显著;而抑制caspase-3并不能完全阻止细胞凋亡.Western-Blot结果显示,姜黄素处理组出现凋亡诱导因子(AIF)蛋白表达与核转位,抑制caspase-3活性后未检测出AIF表达结论:姜黄素可抑制肺成纤维细胞增殖,其诱导大鼠肺成纤维细胞凋亡,可能与线粒体释放AIF有关.     

澳洲茄边碱通过Caspase3蛋白诱导人肺腺癌A549细胞的凋亡作用

运用中药龙葵提取物澳洲茄边碱处理人肺腺癌A549细胞,研究其对A549细胞的抑制及凋亡作用,探讨澳洲茄边碱对肺腺癌的作用机制。通过细胞增殖抑制实验检测不同浓度澳洲茄边碱对A549细胞增殖的影响,采用蛋白印迹法(Western blot)检测凋亡蛋白Caspase3的表达水平,采用流式细胞术测定处理后A549细胞的凋亡水平及细胞周期变化。结果显示,不同浓度澳洲茄边碱均能抑制A549的增殖,呈浓度效应;用不同浓度澳洲茄边碱处理A549细胞24h后,Western blot结果显示,随药物浓度增大,凋亡蛋白Caspase3水解程度增高,对A549凋亡作用明显增强;流式细胞术检测细胞凋亡的结果显示,20μmol·L-1澳洲茄边碱处理A549细胞后,细胞发生明显凋亡,其中早期凋亡细胞比例为25.35%,晚期凋亡细胞比例为11.47%;流式细胞术检测细胞周期的结果显示,20μmol·L-1澳洲茄边碱处理A549细胞后,细胞周期阻滞于G2/M期。本研究结果表明,澳洲茄边碱通过激活细胞凋亡通路中的Caspase3蛋白触发细胞凋亡,同时将A549细胞阻滞在细胞周期的G2/M期,抑制人肺腺癌细胞A549的生长。     

凋亡诱导因子在姜黄素诱导的大鼠肺成纤维细胞凋亡中的作用

目的：研究姜黄素对肺纤维化大鼠肺成纤维细胞增殖、凋亡的影响,探讨凋亡诱导因子（AIF）在肺成纤维细胞凋亡中的作用。方法：将体外培养的肺纤维化大鼠成纤维细胞,分别于不同浓度的姜黄素（5、10、20、40μM）和caspase-3抑制剂Z-DEVD-fmk（20μM）孵育,观测细胞生长状态变化。MTT检测成纤维细胞增殖抑制率;流式细胞仪检测细胞凋亡率;Western-Blot测定凋亡诱导因子（AIF）蛋白表达及核转位结果：流式细胞术检测细胞凋亡,5～40μM姜黄素处理12 h,其凋亡率呈浓度依赖,对照组相比,差异显著;而抑制caspase-3并不能完全阻止细胞凋亡。Western-Blot结果显示,姜黄素处理组出现凋亡诱导因子（AIF）蛋白表达与核转位,抑制caspase-3活性后未检测出AIF表达结论：姜黄素可抑制肺成纤维细胞增殖,其诱导大鼠肺成纤维细胞凋亡,可能与线粒体释放AIF有关。     

细胞周期蛋白依赖激酶Roscovitine 诱导非小细胞肺癌A549 细胞凋亡及 其机制的研究

目的:探讨细胞周期蛋白依赖激酶(CDK)抑制剂Roscovitine(Ros)诱导非小细胞肺癌(NSCLC)A549细胞凋亡及其作用机制。方法:以不同浓度Ros(10μM、20μM、40μM)处理细胞24h,采用Annexin V-PI染色以流式细胞仪检测细胞凋亡,Westernblot法检测胞浆中和线粒体促凋亡蛋白Bax和Bad的表达,流式细胞仪检测线粒体膜电位(MMP)变化。结果:Ros以剂量依赖的方式诱导A549细胞凋亡,同时Bad和Bax在胞浆的含量随着Ros剂量的增加而减少,而在线粒体中却出现相反的结果,线粒体膜电位随Ros剂量的增大而降低。结论:Ros可通过促进Bax和Bad由胞浆向线粒体易位,诱导NSCLC A549细胞由线粒体途径发生凋亡。     

曲古霉素A增强肺癌细胞对放射线敏感性及机制

目的:探讨组蛋白去乙酰化酶抑制剂曲古霉素A(trichostatin A,TSA)增强人非小细胞肺癌(NSCLC)A549对γ-射线敏感性作用及机制。方法:以TSA(0.5μM)预处理细胞18h,再以5Gyγ-射线照射细胞,24h后采用MTT法检测细胞存活率,AnnexinV-PI染色检测细胞凋亡,Westemblot法检测胞浆中和线粒体促凋亡蛋白Bax的表达,流式细胞仪检测细胞线粒体膜电位变化。结果:5Gyγ-射线照射可轻度降低细胞存活率,仅有少量细胞发生凋亡,以TSA预处理再以γ-射线处理细胞,细胞存活率显著下降,凋亡细胞明显增多,伴有线粒体膜电位下降,以及Bax蛋白的激活,表现在线粒体Bax表达较单纯照射组显著增高。结论:TSA通过促进Bax蛋白的活化激活线粒体凋亡途径,增强增强A549细胞对y-射线的敏感性。     

加味苇茎汤及其成分芹菜素对肺腺癌A549细胞增殖、周期与凋亡的影响

探讨加味苇茎汤含药血清及成分芹菜素对肺腺癌A549细胞增殖、细胞周期及凋亡的影响,选用肺癌A549细胞体外培养,在不同浓度含药血清及芹菜素作用下,用MTT法检测细胞生长抑制率、流式细胞术检测细胞周期及凋亡率、hoechst33342染色检测细胞凋亡形态。结果显示,加味苇茎汤含药血清及芹菜素对A549的生长抑制率:24 h分别为31%、34%,48 h分别为45%、53%;流式细胞术检测细胞周期发现A549细胞经加味苇茎汤含药血清处理后主要阻滞在G2期,而芹菜素能明显引起细胞凋亡。荧光显微镜下观察到较典型的细胞凋亡形态甚至细胞脱落,视野内细胞核数明显减少。该研究结果显示,加味苇茎汤含药血清及芹菜素能明显抑制A549细胞的增殖,引起细胞周期阻滞、诱导其凋亡。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.