海藻酸微囊替代DMSO和FBS的STO细胞冷冻保存研究

目的:改进现有的细胞冷冻保存方法,建立一个不含二甲基亚砜(DMSO)和血清(FBS)的高效冷冻保存方法,为细胞治疗等临床实践提供优质细胞.方法:海藻酸微囊包埋鼠胚成纤维细胞(STO细胞)后用不含DMSO和FBS的冷冻保存液进行冷冻保存.设四个对照组:添加10％DMSO和20％FBS的组、仅添加10％DMSO的组、仅添加20％FBS、DMSO和FBS均不添加组.在冷冻前后对各实验组细胞用台盼兰染色,进行细胞计数,计算细胞存活率,同时利用溴乙锭的二聚物(EthD)、钙黄绿素-AM(Calcein-AM)进行染色观察细胞的形态,且进一步验证细胞存活率;解冻复苏后用MTT法评估细胞的增殖速度和生长活力.结果:冷冻保存30天后对各组的细胞数量、细胞存活率、细胞形态和解冻复苏后细胞的生长活力进行比较发现,海藻酸微囊包埋冷冻组的细胞数、细胞存活率、细胞形态和生长活力均与添加DMSO和FBS的组之间无显著性差异,而与其它三个对照组呈显著性差异.结论:使用海藻酸微囊替代DMSO和FBS保存STO细胞,能有效的维持细胞形态、数量、存活率,同时不影响细胞的生长活力,从而建立了一个不含DMSO和FBS的高效冷冻保存方法.     

Zerumbone induced apoptosis in liver cancer cells via modulation of Bax/Bcl-2 ratio

Background  

Zerumbone is a cytotoxic component isolated from Zingiber zerumbet Smith, a herbal plant which is also known as lempoyang. This new anticancer bioactive compound from Z. zerumbet was investigated for its activity and mechanism in human liver cancer cell lines.     

A statistical test that supports a human/chimpanzee clade based on noncoding DNA sequence data

Using the aligned DNA sequence data of Miyamoto et al. and Maeda et al., all noncoding genetic material, and a simple statistical test, we show that a Homo/Pan clade is supported at approximately the 3% level of significance. The method accommodates polymorphism and different evolutionary rates for different sites. All assumptions on which the statistical study is based are made explicit. (See the Note added in proof, which--adding recently published data--improves the significance level to about 1%.      

叶绿素结合蛋白CP24介导光照响应基因StRSM 1调控叶绿素积累

单MYB转录因子成员RSM(RADIALIS-like SANT/MYB)突变,倒置黑暗诱导的叶绿素减少,原因有待确定;为了揭示RSM如何调控叶绿素积累,本研究运用基因工程途径,获得了普通烟草和马铃薯体内抑制和过量表达StRSM 1阳性转化株系,测定了阳性转化株系的叶绿素积累等生理表型;结果显示,普通烟草和马铃薯体内抑制RSM 1表达,显著增加了叶绿素积累,叶色随之加深;RSM 1过量表达,显著减少叶绿素积累,叶色变浅。叶绿素代谢相关基因表达测定结果显示,StRSM 1过量表达增加了黑暗下叶绿素结合蛋白CP24基因的表达,改变了其表达模式。以上结果表明,转录因子StRSM 1响应光照反向调控叶绿素积累,叶绿素结合蛋白CP24参与了StRSM 1对叶绿素积累的调控。结果有助于进一步明确RSM 1如何响应光照和深刻理解RSM 1参与的光照响应。     

Electrical and adaptive properties of rod photoreceptors in bufo marinus. I. Effects of altered extracellular Ca(2+) levels

The effects of altering extracellular Ca(2+) levels on the electrical and adaptive properties of toad rods have been examined. The retina was continually superfused in control (1.6 mM Ca(2+)) or test ringer’s solutions, and rod electrical activity was recorded intracellularly. Low-calcium ringer’s (10(-9)M Ca(2+)) superfused for up to 6 min caused a substantial depolarization of the resting membrane potential, an increase in light-evoked response amplitudes, and a change in the waveform of the light-evoked responses. High Ca(2+) ringer’s (3.2 mM) hyperpolarized the cell membrane and decreased response amplitudes. However, under conditions of either low or high Ca(2+) superfusion for up to 6 min, in both dark-adapted and partially light-adapted states, receptor sensitivity was virtually unaffected; i.e., the V-log I curve for the receptor potential was always located on the intensity scale at a position predicted by the prevailing light level, not by Ca(2+) concentration. Thus, we speculate that cytosol Ca(2+) concentration is capable of regulating membrane potential levels and light-evoked response amplitudes, but not the major component of rod sensitivity. Low Ca(2+) ringer’s also shortened the period of receptor response saturation after a bright but nonbleaching light flash, hence accelerating the onset of both membrane potential and sensitivity recovery during dark adaptation.

Exposure of the retina to low Ca(2+) (10(-9)M) ringer’s for long periods (7-15 min) caused dark-adapted rods to lose responsiveness. Response amplitudes gradually decreased, and the rods became desensitized. These severe conditions of low Ca(2+) caused changes in the dark-adapted rod that mimic those observed in rods during light adaptation. We suggest that loss of receptor sensitivity during prolonged exposure to low Ca(2+) ringer’s results from a decrease of intracellular (intradisk) stores of Ca(2+); i.e., less Ca(2+) is thereby released per quantum catch.

     

Different rewards in female and male flowers can explain the evolution of sexual dimorphism in plants

Insects use floral signals to find rewards in flowers, transferring pollen in the process. In unisexual plants, the general view is that staminate (male) and pistillate (female) flowers obtain conspecific pollen transfers by advertising their rewards with similar floral signals. For female plants lacking food rewards, this can lead to floral mimicry and pollination by deceit. In this study, we challenge this view by presenting evidence for different rewards offered by flowers on females and males, as a mechanism promoting sexual dimorphism in Leucadendron xanthoconus (Proteaceae), a clearly sexually dimorphic shrub. The tiny beetle pollinators Pria cinerascens (Nitidulidae) depend entirely on the plants they pollinate for survival and reproduction. Male flowers provide mating and egglaying sites, and food for adults and larvae. Female flowers lack nectar and function to shelter pollinators from rain. Their flower heads have cup‐shaped display leaves, and are more closed than are those in males. On rainy days, flowers on females received 30% more visits than did flowers on males, and 90% more than they did on sunny days. When we removed display leaves in females, intact flower heads received 14 times more P. cinerascens visits than did manipulated flower heads, indicating that the cup shape attracts the beetles. In both sexes, having many flowers increased the probability of visits and the number of P. cinerascens visiting a plant. In males, the number of larvae was positively correlated with floral‐display size, while in females, seed set (pollen transfers) showed no relationship with floral‐display size. Ninety‐five per cent of the ovules received pollen and 52% matured into seeds. We explain the sexual dimorphism in L. xanthoconus as a result of an intimate partnership with P. cinerascens pollinators, in conjunction with a rainy climate. Pollinators favour large male floral displays, because they offer a reliable food source for adults and larvae. Frequent rains drive the P. cinerascens to leave males in search of the protection offered by females. Because females offer shelter, an essential resource that is not offered by male plants, they receive sufficient pollen independent of their floral‐display size. This pollination system promotes the evolution of sexually dimorphic floral signals, guiding pollinators to different rewards in male and female flowers. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 85 , 97–109.     

Mycotoxins of Alternaria alternata produced on ceiling tiles

The production of mycotoxins by Alternaria alternata in cellulosic ceiling tiles was examined with thin-layer chromatography and high-performance liquid chromatography procedures. Alternariol and alternariol monomethyl ether were found in ceiling tile extracts, whereas extracts of control rice cultures of all three isolates produced these mycotoxins plus altenuene and altertoxin I. Extensive fungal growth and mycotoxin production occurred in the ceiling tiles at relative humidities of 84–89% and 97%. Received 28 May 1997/ Accepted in revised form 06 October 1997     

海藻酸微囊替代DMSO和FBS的STO细胞冷冻保存研穷

目的：改进现有的细胞冷冻保存方法,建立一个不舍二甲基亚砜（DMSO）和血清（FBS）的高效冷冻保存方法,为细胞治疗等临床实践提供优质细胞。方法：海藻酸微囊包埋鼠胚成纤维细胞（STO细胞）后用不含DMSO和FBS的冷冻保存液进行冷冻保存。,设四个对照组：添加10％DMSO和20％FBS的组、仅添加10％DMSO的组、仅添加20％FBS、DMSO和FBS均不添加组。在冷冻前后对各实验组细胞用台盼兰染色,进行细胞计数,计算细胞存活率,同时利用溴乙锭的二聚物（EthD）、钙黄绿素-AM（Calcein—AM）进行染色观察细胞的形态,且进一步验证细胞存活率;解冻复苏后用MTT法评估细胞的增殖速度和生长活力。结果：冷冻保存30天后对各组的细胞数量、细胞存活率、细胞形态和解冻复苏后细胞的生长活力进行比较发现,海藻酸微囊包埋冷冻组的细胞数、细胞存活率、细胞形态和生长活力均与添加DMSO和FBS的组之间无显著性差异,而与其它三个对照组呈显著性差异。结论：使用海藻酸微囊替代DMSO和FBS保存STO细胞,能有效的维持细胞形态、数量、存活率,同时不影响细胞的生长活力,从而建立了一个不含DMS0和FBS的高效冷冻保存方法。     

几种旱生灌木种子萌发对干旱胁迫的响应

以不同渗透势的PEG(6 0 0 0 )溶液 (- 0 .3～ - 2 .7MPa)为模拟干旱胁迫条件 ,研究了柠条 (Caraganakorshinskii)、花棒 (Hedysarumscoparium)和白沙蒿 (Artemisiasphaerocephala)种子发芽、幼苗生长和累积吸水率对干旱胁迫的响应 ,讨论了参试种子发芽特性、初生根长度与幼苗建植成活率的关系 .结果表明 ,干旱胁迫下柠条发芽率最高 ,其次为花棒 ,白沙蒿最低 . - 0 .3～ - 0 .6MPa渗透势下白沙蒿发芽 10d的初生根长度显著大于花棒和柠条 (P <0 .0 5 ) ,三者的测量值依次为 7.9、4 .5和 3.1cm .干旱胁迫条件下参试种子发芽率与种子 72h累积吸水率均呈极显著的正相关 (P <0 .0 1) .     

Myrmecophilous butterflies utilise ant–treehopper associations as visual cues for oviposition

1. Selection of a safe oviposition site is important for herbivorous insects whose immature stages have limited mobility. Female herbivores rely on environmental cues for this choice, and presence of natural enemies or mutualistic partners may be important in this process. 2. Some butterflies have mutualistic interactions with ants (myrmecophily), in which caterpillars offer a nutritional liquid and gain protection against natural enemies. Participants in butterfly–ant mutualisms may utilise signals to initiate interactions, but the use of visual cues by ovipositing myrmecophilous butterflies remains uncertain. 3. Larvae of facultatively myrmecophilous Parrhasius polibetes (Lycaenidae) feed on Schefflera vinosa, and females prefer to oviposit near aggregations of the ant‐tended treehopper Guayaquila xiphias, where caterpillars survive better due to increased ant attendance. Given the conspicuousness of ant–treehopper associations, it was investigated whether butterflies use them as visual cues for oviposition and, if so, which participants of the association are used as cues: ants, treehoppers, or both. 4. Experiments using dried insects on paired branches revealed that females visually recognise ants and ant–treehopper associations, using them for egg‐laying decisions. However, presence of a treehopper aggregation alone had no effect on oviposition choices. 5. This is a first insight into the importance of visual discrimination for ovipositing myrmecophilous butterflies. The results show that facultative mutualisms can be important enough to promote a behavioural adaptation (visual detection of ants) reinforcing the interaction. Our research highlights the importance of the behavioural interface within complex multispecies systems.