环境激素DBP对拟南芥体外培养叶片超微结构的影响

报道了酞酸酯类化合物DBP对拟南芥离体培养叶片超微结构的影响。在DBP(1.0mg·L-1)处理第3天即观察到拟南芥叶片叶绿体的超微结构受到破坏。不同浓度的DBP长期处理植株(40d)出现叶片白化、节间短缩等异常现象。在0.01mg·L-1DBP处理40d植株的叶细胞中,叶绿体出现解体,破碎部分呈颗粒状,散落其间,但细胞结构完整;0.1mg·L-1DBP处理后引起叶绿体的进一步解体,叶绿体中类囊体基粒和片层结构解体,细胞出现空洞现象,其它细胞器很少观察到;1.0mg·L-1DBP和2.0mg·L-1DBP处理植株叶片叶绿体中类囊体基粒和片层结构破碎,叶绿体结构也解体,细胞中其它细胞器数目极少。     

胡萝卜愈伤组织细胞中与重力相关酯酶（grEST1和grEST2）的鉴别

用水平回转器改变重力对细胞的作用方式并用以模拟微重力对胡萝卜细胞的影响。经硼酸缓冲液提取,聚丙烯酰胺凝胶生趣平板电泳图谱显示８条酯酶带。其中,只有两种酯酶活力受回转处理的影响。     

模拟微重力诱导的细胞微丝变化影响COL1A1启动子活性

细胞骨架系统是细胞内的重力感受系统。已知微重力导致的细胞形态、功能、信号传导等多种变化均与细胞骨架系统变化有关,但微重力对相关基因调控的影响知之甚少。本研究以构建的基因工程细胞株（EGFP-ROS）为对象,以回转器模拟微重力效应,利用增强型绿色荧光蛋白（enhanced green fluorescence protein,EGFP）荧光半定量和细胞微丝荧光染色分析技术,探讨回转模拟微重力条件下,细胞微丝系统对Ⅰ型胶原α1链基因（collagen type Ialpha chain 1 gene,COL1A1）启动子活性的影响。空间飞行和回转模拟微重力后,细胞微丝解聚、张力纤维减少,表明微重力可降低细胞微丝结构的有序性,诱导细胞骨架重排。适合剂量的细胞松弛素B处理EGFP-ROS细胞诱导微丝骨架解聚,同时导致COL1A1启动子活性增加,细胞荧光强度增强,并呈现剂量依赖性。因此,一定程度的细胞微丝系统破坏将导致COL1A1启动子活性的增强,证明细胞微丝骨架系统参与了微重力对COL1A1启动子活性调节,且在微重力信号传导中起重要作用。     

模拟微重力效应通过微丝骨架调控成骨细胞BMP2-Smad信号

细胞微丝骨架在力信号传导和基因表达调控中起重要作用。为了研究微丝骨架在模拟微重力效应调控成骨细胞BMP2-Smad信号中的作用,作者通过构建反映Smad活性的报告基因载体转染MC3T3-E1细胞,并通过报告基因活性分析、Western blot等方法检测了微丝骨架解聚剂和回转模拟微重力效应对BMP2诱导Samd磷酸化、核质分布和转录活性的作用。结果显示,构建的报告基因载体在成骨细胞中正确表达并响应BMP2;破坏微丝骨架会抑制BMP2诱导的Smad1/5/8蛋白磷酸化、入核及转录活性;回转抑制Smad1/5/8磷酸化、入核及其转录活性,而微丝骨架稳定剂可对抗回转的抑制作用。因此,认为回转模拟微重力效应可通过解聚微丝骨架抑制BMP2-Smad信号传导。     

模拟微重力诱导PC12细胞衰老的初步探讨

目的：构建模拟微重力条件下PC12细胞的培养体系,探讨模拟微重力对PC12细胞衰老的影响。方法：用Cytodex-3型微载体作为PC12细胞的贴附载体,旋转细胞培养系统所提供10-2g的微重力环境进行模拟微重力条件下的细胞培养。在倒置显微镜下观察PC12细胞的生长情况;用扫描电镜观察PC12细胞超微结构的变化;衰老相关β半乳糖苷酶（SA-β-gal）特异性染色对衰老的PC12细胞进行评估。结果：光镜下模拟微重力培养的PC12细胞表现出类衰老细胞的形态,扫描电子显微镜下观察发现其微绒毛增多。SA-β-gal染色的结果显示在模拟微重力的作用下,PC12细胞SA-β-gal的活性升高。结论：模拟微重力可以引起PC12细胞衰老样的形态变化,以及SA-β-gal的活性升高。     

回转器模拟微重力刺激对不同品系稻田鱼腥藻代谢特性的影响

以稻田鱼腥藻空间搭载克隆株 (AoSR1 6)、回复再搭载克隆株 (AoSR1 6 1 7)和原始出发株 (AnabeanaoryzaHB2 3)为材料 ,通过回转器模拟微重力刺激实验 ,对不同品系稻田鱼腥藻的微重力生物学效应进行了分析。结果发现 ,模拟微重力刺激对稻田鱼腥藻不同品系均表现出一定的生长刺激效应 ,尤为空间飞行后的克隆株更为明显。比较三个品系在微重力刺激下的光合与呼吸活性 ,原始出发株的光合与呼吸活性明显高于空间搭载株。在回转器培养情况下 ,具有高固氮酶活性的克隆株 (AoSR1 6和AoSR1 6 1 7)所固定的氮 ,除了用于藻细胞正常的生命活动外 ,主要用于其生长增强效应 ,而藻胆蛋白累积量和氨分泌量较之对照培养时要少得多。对回转器培养后的稻田鱼腥藻进行单克隆分离 ,结果没有出现类似于空间搭载实验的性状分离现象。     

模拟微重力诱导PC12 细胞衰老的初步探讨

目的:构建模拟微重力条件下PC12细胞的培养体系,探讨模拟微重力对PC12细胞衰老的影响。方法:用Cytodex-3型微载体作为PC12细胞的贴附载体,旋转细胞培养系统所提供10-2g的微重力环境进行模拟微重力条件下的细胞培养。在倒置显微镜下观察PC12细胞的生长情况;用扫描电镜观察PC12细胞超微结构的变化;衰老相关β半乳糖苷酶(SA-β-gal)特异性染色对衰老的PC12细胞进行评估。结果:光镜下模拟微重力培养的PC12细胞表现出类衰老细胞的形态,扫描电子显微镜下观察发现其微绒毛增多。SA-β-gal染色的结果显示在模拟微重力的作用下,PC12细胞SA-β-gal的活性升高。结论:模拟微重力可以引起PC12细胞衰老样的形态变化,以及SA-β-gal的活性升高。     

锰胁迫对垂序商陆叶片形态结构及叶绿体超微结构的影响

以超富集植物垂序商陆为实验材料,通过温室实验,研究不同锰处理条件下垂序商陆叶片受害症状、叶片形态结构和叶绿体超微结构的变化。当生长介质中锰供应水平为1 000 μmol?L-1时,垂序商陆生长良好,叶片未表现出受害症状,其形态结构和超微结构也没有明显的变化;当锰供应水平为5 000 μmol?L-1时,叶片开始出现褪绿现象,叶肉细胞排列疏松,栅栏组织细胞膨胀, 叶绿体数量减少,外膜部分解体,类囊体膨胀, 基质片层扭曲, 淀粉粒颗粒变小;当锰供应水平为12 000μmol?L-1时,叶片发生大量的褪绿现象,伤害率高达87.33%,并且伤害率随着锰胁迫时间的延长明显升高,叶肉细胞发生扭曲变形,叶绿体皱缩,外膜解体,类囊体空泡化加剧,基质片层严重扭曲,基粒排列紊乱甚至模糊成絮状,嗜锇颗粒增多。 尽管在高锰浓度胁迫条件下植物叶肉细胞及叶绿体超微结构出现一定程度的毒害特征,但垂序商陆仍能完成部分功能,维持其个体的生长,这进一步表明超富集植物垂序商陆具有极强的锰耐性。     

模拟微重力对恶性胶质瘤细胞U87 影响的实验研究

目的:探讨模拟微重力(Modeled microgravity,MMG)对恶性胶质瘤细胞U87形态、生长增殖、基质金属蛋白酶-2、-9(Matrix metalloproteinase 2,MMP-2、Matrix metalloproteinase 9,MMP-9)及神经胶质酸性蛋白(Glial fibrillary acidic protein,GFAP)表达的影响。方法:常规培养U87细胞,传代培养3代后分为两组,正常重力组(Normal gravity,NG组)及模拟微重力干预组(Modeled microgravity,MMG组),相应条件下培养24 h,48 h和72 h。倒置显微镜观察U87细胞形态改变,细胞计数法及四甲基偶氮唑盐微量酶反应比色法(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT法)检测模拟微重力干预和干预后U87细胞生长增殖情况;Western Blot检测不同培养时间后胶质瘤细胞U87 MMP-2、MMP-9及GFAP蛋白的表达情况。结果:模拟微重力条件下培养72 h后,U87细胞轮廓不规则,边缘圆钝,触角变少;模拟微重力条件下,U87细胞生长速度明显减慢,并且经模拟微重力干预48 h和72 h后的胶质瘤细胞经传代再培养,其增殖率也明显低于正常重力组;同时,U87细胞MMP-2及MMP-9蛋白表达水平与模拟微重力处理时间呈时间依赖性下降,而GFAP蛋白表达水平则呈时间依赖性升高。结论:模拟微重力影响U87细胞的形态、生长及表型。     

Mg~(2+)和SeO_3~(2-)对鸡胚软骨细胞受模拟微重力不良影响的拮抗

在回转模拟微重力条件下 ,研究了鸡胚负重软骨细胞骨架的微管系统和碱性磷酸酶活性两项指标的变化 ,以及 1mg/L亚硒酸钠和 5mmol/LMg2 + 对这些指标的影响 .流式细胞仪对微管含量的测定显示回转后微管蛋白含量的减少 ,说明微管系统受到不良影响 .碱性磷酸酶活性比对照组明显降低 ,表明模拟微重力能降低软骨细胞的钙化能力 .如果在回转前加入SeO2 -3 和Mg2 + ,发现SeO2 -3 可以在一定程度上拮抗模拟微重力引起的微管蛋白及碱性磷酸酶活性改变 ,而Mg2 + 基本上可以完全拮抗模拟微重力对这两项指标的不良影响     

A COMPARATIVE ANALYSIS OF CELL MORPHOLOGY IN MATURING TISSUES OF ROOTS FROM SHOCKED AND UNSHOCKED BULBS OF ALLIUM CEPA L.

A shock pressure pulse of 60 pounds per square inch (psi; 4.22 kg/cm²) effectively inhibited root growth of onion bulbs. Morphological changes observed after shock included a reduction in cell number in transverse section, a decrease in cell length, a decrease in cell volume, and an increase in cell cross-sectional area. Mitotic activity was consistently increased one day after shock, and this may have accounted for the increased cell number per millimeter of root tip segment 8 days after shock. The development of the tissues appeared normal after shock exposure; however, the tissue response to pressures seems to depend on whether they are exposed to prolonged confining pressures or a brief pressure pulse. Responses which were unique to shock treatment include a decrease in cell number in transverse section, reduced cell volume, and increased radial enlargement of the cell. These responses have not been observed under prolonged pressure treatment.     

CPPU对猕猴桃果实组织发育及内源细胞分裂素含量的影响

对经生长调节剂ＣＰＰＵ处理的猕猴桃果帝进行了组织学及内源细胞分裂素含量变化的研究。结果表明：处理后的果帝细胞分裂时期延长,细胞的数目增多,体积增大;处理果实中,内源细胞分裂素的含量在果肉细胞分裂盛期及其之后都有所增高。     

锰对作业工人血液流变学的影响及清栓酶的疗效观察

本文对１０３例锰作业工人测定了血液流变学中的七项指标,结果表明：全血粘度（高切、低切）、血浆粘度、红细胞压积、纤维蛋白原、红细胞电泳、血小板粘附、血沉等项指标,均明显高于正常对照组（Ｐ＜０．０１）,证实锰作业工人普遍存在高粘滞血症,为临床治疗锰中毒开辟新的途径提供了依据。对其中的５２例锰中毒（包括症状较重的观察对象）,用清栓酶治疗后,复查其七项指标与治疗前比较,均有明显下降（Ｐ＜０．０１）,其症状与体征得以改善,进一步说明清栓酶是治疗锰中毒的理想药物之一。     

EFFECTS OF CORTICOSTEROIDS ON THE BIOCHEMICAL MATURATION OF RAT BRAIN: POSTNATAL CELL FORMATION

(1) Treatment with cortisol acetate (0.2 mg daily during the first 4 days after birth) reduced the rate of growth in the rat: at 35 days of age the body weight was reduced by 50 per cent and the brain weight, depending on the region, by up to 30 per cent. (2) In the brain the normal increase in cell number was severely inhibited during the period of cortisol treatment; this resulted in a final deficit in cell number of about 20 per cent in the cerebrum and 30 per cent in the cerebellum. (3) To determine whether cortisol affected primarily cell formation or cell destruction the labelling of brain DNA was studied 1 h after a subcutaneous injection of 20 Ci/100 g [2-¹⁴C]thymidine. In the controls the amount of labelled DNA increased by a factor of two in the cerebrum and seven in the cerebellum during the period 2-13 days, and it decreased to 40 and 27 per cent of the peak values in the cerebrum and cerebellum respectively in the following 7 days. The results indicated that mitotic activity is higher in the cerebellum than in the cerebrum in the 2nd week of life. It would appear that in the cerebrum appreciable cell death accompanies new cell formation, especially during the period 13-35 days of age. (4) Cortisol treatment affected cell division rather than cell destruction in the brain since it strongly inhibited the incorporation of [2-¹⁴C]thymidine into DNA. The inhibition was severe during the period of treatment but it did not result in a lasting fall in mitotic activity. At the age of 13 days the amount of labelled DNA formed approached the normal level and it was twice that in controls at 20 days, indicating a tendency for compensating cell deficit by an accelerated mitotic activity. Nevertheless, massive cell proliferation ceased at about the same age as in normals; the labelling of DNA decreased markedly between 13 and 20 days after birth, and the DNA content did not increase after the age of 20 days. (5) In contrast to the marked effect on cell number, cortisol treatment did not influence significantly the maturational changes related to average cell size (DNA concentration) or the chemical composition of cells (RNA/DNA and protein/DNA).     

METHODS FOR DETERMINING THE PROLIFERATION KINETICS OF CELLS BY MEANS OF 5-BROMODEOXYURIDINE

After treatment of cells with 5-bromodeoxyuridine (BUdR), the percentage of completely BUdR-labelled interphase nuclei is greater the longer the BUdR treatment. The labelling effect is visible after staining with the fluorochrome 33258 Hoechst and with Giemsa. Various formulae and a nomogram are presented by means of which the percentage of cells in S period, duration of the S period and the whole cell cycle can be determined by examination of a single preparation or by comparison of several preparations. The methods are tested using cell cultures of Microtus agrestis and compared with autoradiographic methods after labelling with ³H-thymidine.     

细叶黄芪叶肉原生质体发育早期细胞壁再生的研究

采用透射电镜术、电镜多糖细胞化学染色、细胞壁荧光染色以及香豆素抑制细胞壁再生等方法,对细叶黄芪（Ａｓｔｒａｇａｌｕｓｍ ｅｌｉｌｏｔｏｉｄｅｓ ｖａｒ．ｔｅｎｕｉｓ）叶肉原生质体细胞壁的再生及其化学特点进行了研究。结果表明,离体培养２４ 小时的原生质体表面产生一些突起小泡,有时可见少量纤维组分的形成。培养３ 天时这种纤维组分明显增多。至５ 天时可清楚看到再生壁是由纤维和颗粒构成。六亚甲四胺银染色证明它们都是由多糖组分组成的。另外,培养３６ 小时的原生质体有相互粘连的现象。电镜观察、荧光染色及香豆素处理的研究表明粘连与再生壁的形成有关。根据上述观察结果,对原生质体再生壁的结构及其化学性质等问题进行了讨论     

HSF2 mRNA在热应激和大剂量11酸睾酮诱导恒河猴生精细胞凋亡中的表达变化

为了探讨HSF2 mRNA在热应激和超生理剂量睾酮诱导恒河猴生精细胞凋亡中的表达变化,我们建立了手术诱导单侧隐睾和注射大剂量11酸睾酮（TU）恒河猴动物模型,应用3′末端标记分析（TUNEL）和原位杂交方法,检测睾丸细胞的凋亡信号和HSF2的表达变化。TUNEL结果显示热应激和超生理剂量睾酮能够诱导生精细胞出现凋亡信号,它分别于处理后第5天和第30天达到最强,表明热应激和睾酮干扰精子发生可能是通过生精细胞凋亡的方式来实现的。HSF2 mRNA水平在生精细胞凋亡早期（凋亡信号达到最强以前）略有降低,而在凋亡高峰期之后其表达急剧下降。Hsf2基因与我们以前研究的Hsp70-2基因的表达具有时间上的相关性,表明HSF2蛋白可能调控Hsp70-2基因的表达,而且HSF2可能通过多种方式影响精子的发生以及抑制生精细胞的凋亡。     

INCORPORATION OF TRITIATED THYMIDINE DURING VARIOUS STAGES OF LEAF DEVELOPMENT OF XANTHIUM PENNSYLVANICUM

³H-thymidine was incorporated into leaf tissue of Xanthium pennsylvanicum during the stage of active cell division, during cellular differentiation, and into mature cells. Incorporation into nuclear DNA was high in the early stages of development. No nuclear incorporation was found after cessation of cell division. However, significant incorporation could be demonstrated in cytoplasm of differentiating and mature cells. Depending upon the time of growth in the radioisotope and the time of growth after treatment, ³H-thymidine, or its metabolized fraction, was incorporated into the secondary wall depositions of epidermal cells, mesophyll parenchyma cells, xylem cells, and chloroplasts. Autoradiographic technique and liquid scintillation spectrometry were used in these studies. The significance of ³H-thymidine incorporation into various organelles is discussed in relation to cell metabolism and its regulation during leaf development.     

INDUCTION OF KERATINOCYTE PROLIFERATION BY A SHORT TREATMENT WITH KERATINOCYTE-CONDITIONED MEDIUM

The effect of a short treatment with keratinocyte-conditioned medium (KCM) on the growth of normal human epidermal keratinocytes was investigated. Serum-free MCDB153 medium was conditioned by keratinocytes for 24h after plating. Following attachment to uncoated plastic surfaces (4h after plating), cells were exposed for 20h to KCM. After 10 days of culture in MCDB153 medium, an increase of about six-fold in cell number was observed in KCM-treated plates over controls, indicating that a short treatment with KCM is sufficient to induce cell proliferation. The effect of addition of KCM at different times after plating was also evaluated: KCM treatment resulted to exert its maximum effect on cell proliferation, when performed immediately after the completion of attachment of cells to the surface of the dish. Mitogenic activity present in KCM is not inhibited by heparin sulphate. The kinetics of accumulation of this early secreted growth-stimulating activity showed that a plateau is reached within 24h of conditioning. These data suggest that this mitogenic activity should not be amphiregulin. The observation that, following KCM treatment, the majority of cells is able to incorporate [³H]-thymidine as compared to controls suggests that the observed final increase in cell number is due to an increase in the number of cycling cells rather than to a shortening of the cell cycle.     

黄芩甙对肝癌细胞BEL-7402形态学的影响

目的观察黄芩甙对肝癌细胞BEL-7402凋亡的影响,同时观察对肝癌细胞形态及超微结构、线粒体超微结构、线粒体膜电位和细胞内Ca^2＋的影响,探讨线粒体损伤在黄芩甙诱导肝癌细胞凋亡中的作用及可能的机制。方法应用细胞培养技术培养肝癌细胞BEL-7402,光镜、倒置显微镜、扫描电镜、透射电镜观察细胞形态及超微结构的变化尤其是线粒体的变化,应用流式细胞仪检测细胞凋亡百分率及线粒体膜电位、细胞内Ca^2＋的改变,免疫组化法检测细胞Bcl-2、Pax蛋白表达。结果黄芩甙诱导肝癌细胞BEL-7402凋亡呈剂量依赖关系,细胞形态、超微结构及线粒体超微结构出现明显改变,降低肝癌细胞线粒体膜电位,使细胞内Ca^2＋增加,细胞Pax表达增加,广泛分布于胞核和胞质中,Bcl-2表达减少。结论黄芩甙诱导肝癌细胞BEL-7402凋亡,线粒体损伤在黄芩甙诱导肝癌细胞凋亡中起重要作用,其机制可能为抑制肝癌细胞Bcl-2蛋白表达,促进Pax蛋白表达及细胞内Ca^2＋增加,激发线粒体膜通透性转运孔开放,线粒体跨膜电位降低,使肝癌细胞凋亡。