太白银莲花皂苷B对胶质瘤细胞生长抑制的研究

目的:太白银莲花皂苷B(Anemone taipaiensis saponin B)是第一次从太白银莲花中经过系统化学分析和分离鉴定的皂苷之一,所以它的生物学效应目前仍然不清楚。在本研究中,我们首次体外研究太白银莲花皂苷B对胶质瘤细胞系的生物学效应,观察它对胶质瘤细胞增殖的的抑制作用。方法:采用四甲基偶氮唑蓝(MTT)法测定太白银莲花皂苷B对胶质瘤细胞生长曲线的影响,Hoechst 33342细胞核染色后荧光显微镜观察,采用光学显微镜观察细胞的形态学变化。结果:MTT实验结果显示太白银莲花皂苷B对胶质瘤细胞U87MG和U251MG有强烈的生长抑制作用,且具有剂量依赖性,应用SPSS18.0统计软件得出太白银莲花皂苷B对U87MG细胞72 h的抑制浓度为IC25=5.2μmol/L,IC50=6.7μmol/L and IC75=8.7μmol/L,U251细胞的抑制浓度为IC25=6.2μmol/L,IC50=7.9μmol/L and IC75=10.5μmol/L。Hoechst 33342细胞核染色荧光显微镜观察以及光学显微镜下细胞形态观察显示出典型的凋亡细胞形态学特征,经过皂苷B处理后,细胞皱缩成圆球形,细胞核碎裂或者致密浓染,向核膜边缘聚集,染色质浓缩为半月状、车轮状或者马蹄状,凋亡小体出现。这些特征在24 h时更明显。结论:体外实验初步显示,太白银莲花皂苷B对U87MG和U251MG细胞具有明显的增殖抑制作用,并具有促凋亡作用。     

Dynamics of vacuoles and actin filaments in guard cells and their roles in stomatal movement

Vacuoles and actin filaments are important cytoarchitectures involved in guard cell function. The changes in the morphology and number of vacuoles and the regulation of ion channel activity in tonoplast of guard cells are essential for stomatal movement. A number of studies have investigated the regulation of ion channels in animal and plant cells; however, little is known about the regulating mechanism for vacuolar dynamics in stomatal movement. Actin filaments of guard cells are remodelling with the changes in the stomatal aperture; however, the dynamic functions of actin filaments in stomatal movement remain elusive. In this paper, we summarize the recent developments in the understanding of the dynamics of actin filaments and vacuoles of guard cells during stomatal movement. All relevant studies suggest that actin filaments might be involved in stomatal movement by regulating vacuolar dynamics and the ion channels in tonoplast. The future study could be focused on the linker protein mediating the interaction between actin filaments and tonoplast, which will provide insights into the interactive function of actin and vacuole in stomatal movement regulation.     

木材细胞壁力学研究进展

基于细胞水平的微观力学研究是探索木材复杂力学行为本质的核心内容。该文系统介绍木材细胞壁力学性能实验表征技术及力学模型的发展,重点说明了我国在这方面取得的进展和成果,在此基础上,明确指出细胞壁力学在现阶段存在的不足,并对其发展方向进行详细阐述,以期丰富和推动我国木材科学理论体系的发展,促进纸浆造纸、纺织材料、复合材料等相关领域的研究进展。     

小麦叶片磷酸酯酶生化特性的研究

采用分光光度法对小麦叶片磷酸酯酶的生化动力学特性进行了研究.实验结果表明:叶片磷酸酯酶水解pNPP的Km值为6.554 mm o l/L,Vm ax为0.774×103U/(m g prote in).该酶的最适pH为6.8,保温10 m in的最适温度为45℃,在50、60、70℃保温时,该酶活力丧失50%所需的时间分别为12、3.75和1.8 m in.不同的金属离子及络合剂对该酶活力有影响,M g2 和C a2 对该酶有激活作用,Zn2 、Cu2 和M n2 对酶活力则有不同程度的抑制作用.EDTA、磷酸氢二钠和偏钒酸铵对该酶活力也有抑制作用.利用L inew eaver-Burk作图法判定抑制类型分别为非竞争性、竞争性和非竞争性抑制,抑制常数分别为5.87、2.47和8.2μm o l/L.     

翅碱蓬CMO基因克隆、测序及植物表达载体的构建

从翅碱蓬叶片中提取总RNA,根据相关同源序列设计引物,使用反转录试剂盒进行RT-PCR,扩增得到翅碱蓬胆碱单加氧酶（CMO）cDNA,进行PCR产物测序;然后将CMOcDNAT-A克隆至pMD18-T-simple载体上,经测序正确后亚克隆至pBI121植物表达载体,进行酶切鉴定及克隆测序。     

乳酸菌在肠道内黏附定殖的研究进展

乳酸菌是一类无芽孢的革兰氏阳性菌,能利用碳水化合物发酵产酸。由于其具有安全高效,易获取的优势,在食品、医疗及动物生产方面得到广泛运用。在动物肠道内,它利用糖类物质,生成酸性代谢物,降低肠道pH,改善肠道微生物环境。要实现乳酸菌在动物体内的良好定殖,首先要实现黏附,而黏附过程主要分为特异性结合与非特异性结合。黏附机制的相关研究中黏附素理论被普遍认可,包含了粘附蛋白学说、磷璧酸学说及细菌表面分子学说。本文就乳酸菌的黏附相关因子,体外黏附模型及黏附的影响因素做了阐述。     

中华蜜蜂热休克蛋白70的克隆、表达及中蜂囊状幼虫病毒感染后表达差异分析

[目的]本研究旨在明确中华蜜蜂Apis cerana cerana热休克蛋白70 (Heat shock protein 70,HSP70)与中蜂囊状幼虫病毒(Chinese sacbrood virus,CSBV)的关系.[方法]首先根据NCBI上已公布的中华蜜蜂HSP70的全基因(NO.MH122655.1)序列,设计特异性引物,提取3日龄中华蜜蜂幼虫总RNA,通过RT-PCR方法获得HSP70基因,并通过生物信息学软件进行分析;其次将其克隆到原核表达载体pET-32a中,转化入大肠杆菌BL21(DE3)进行蛋白表达,并制备重组蛋白,将纯化的重组蛋白免疫小鼠制备多克隆抗体;最后利用制备多克隆抗体作为一抗,通过Western-blot方法检测CSBV感染中华蜜蜂前后HSP70在幼虫体内的变化.[结果]成功获取中华蜜蜂HSP70蛋白的全基因,其核苷酸序列全长为1833 bp,生物信息学分析后,发现其编码蛋白存在7个抗原表位.利用原核表达系统,成功获得大小约87 ku的重组蛋白HSP70,免疫小鼠后获得多克隆抗体,经Western-blot分析能与标签蛋白发生特异性反应;对感染CSBV前后蜜蜂体内HSP70蛋白检测发现感染后HSP70表达水平显著提高.[结论]通过原核表达系统,成功表达中华蜜蜂HSP70基因和HSP70多克隆抗体,并证实CSBV感染后影响中华蜜蜂体内HSP70蛋白表达量的改变,为深入研究HSP70功能提供了帮助.     

A New Species of the Genus Tylototriton （Urodela： Salamandridae） from Northeastern Hunan Province,China

We describe a new species from the family Salamandridae from northeastern Hunan Province of China based on morphological and molecular genetic evidences.The new species,Tylototriton liuyangensis,is a member of the T.asperrimus group,and shares a number of similarities with T.wenxianensis and T.broadoridgus.The dorsal surface of the new Tylototriton species is completely black,with nodule-like warts distributed evenly along the lateral margin of dorsal body,and there is no transverse striae between the warts.There are several diagnostic characters of the new species that differ from those of T.wenxianensis and T.broadoridgus as follows：（1）the interorbital space（IOS）is comparatively wide;（2）there is a bony ridge present on the dorsal head surface,running along the upper eyelids;（3）the space between axilla and groin（AGS）is comparatively large,with the mean male AGS approximately equal to 50%of the snout-vent length（SVL）and the mean female AGS being 53%of SVL;and（4）there is a small genital armature on the anterior angle of labial folds present inside the anal fissure of adult males during the breeding season.The result from the molecular phylogenetic analysis of the genus Tylototriton（including the type specimen of the new species）based on the complete DNA sequence of the mitochondrial ND2 gene（1035 bp）indicated that the new species was similar to T.wenxianensis,T.dabienicus,and T.broadoridgus,but formed an independent clade.This result was consistent with the morphological analysis above,supporting that the Liuyang Population represented a distinct species.     

Effects of elevated ozone concentration on methane emission from a rice paddy in Yangtze River Delta,China

Few investigations have been made on the impact of elevated ozone (O₃) concentration on methane (CH₄) emission from rice paddies. Using open‐top chambers in situ with different O₃ treatments, CH₄ emissions were measured in a rice paddy in Yangtze River Delta, China in 2007 and 2008. There were four treatments applied: charcoal‐filtered air (CF), nonfiltered air (NF), and charcoal‐filtered air with different O₃ additions (O₃‐1 and O₃‐2). The mean O₃ concentrations during the O₃ fumigation were 19.7, 22.6, 69.6 and 118.6 ppb in 2007 and 7.0, 17.4, 82.2 and 138.3 ppb in 2008 for treatments CF, NF, O₃‐1 and O₃‐2, respectively. The rice yields, as compared with CF, were reduced by 32.8% and 37.1%, 58.3% and 52.1% in treatments O₃‐1 and O₃‐2 in 2007 and 2008, respectively. The diurnal patterns of CH₄ emission varied temporally with treatments and there was inconsistence in diurnal variations in CH₄ emissions from the paddy field. The daily mean CH₄ emissions were significantly lower in treatments O₃‐1 and O₃‐2 than those in treatments CF and NF. Compared with CF treatment, CH₄ emissions from the paddy field were decreased to 46.5% and 38.3%, 50.6% and 46.8% under treatments O₃‐1 and O₃‐2 in the whole growing seasons of 2007 and 2008, respectively. The seasonal mean CH₄ emissions were negatively related with AOT40 (accumulative O₃ concentration above 40 ppb; P < 0.01 in both years), but positively related to the relative rice yield (reference to CF; P < 0.01 in 2007 and P < 0.001 in 2008), aboveground biomass (P < 0.01 in both years) and underground biomass (P < 0.01 in 2007 and P < 0.05 in 2008). The decreased CH₄ emission from the rice paddy due to an increased O₃ exposure might partially mitigate the global warming potential induced by soil carbon loss under elevated O₃ concentrations.     

Seasonal hysteresis of net ecosystem exchange in response to temperature change: patterns and causes

Understanding how net ecosystem exchange (NEE) changes with temperature is central to the debate on climate change‐carbon cycle feedbacks, but still remains unclear. Here, we used eddy covariance measurements of NEE from 20 FLUXNET sites (203 site‐years of data) in mid‐ and high‐latitude forests to investigate the temperature response of NEE. Years were divided into two half thermal years (increasing temperature in spring and decreasing temperature in autumn) using the maximum daily mean temperature. We observed a parabolic‐like pattern of NEE in response to temperature change in both the spring and autumn half thermal years. However, at similar temperatures, NEE was considerably depressed during the decreasing temperature season as compared with the increasing temperature season, inducing a counter‐clockwise hysteresis pattern in the NEE–temperature relation at most sites. The magnitude of this hysteresis was attributable mostly (68%) to gross primary production (GPP) differences but little (8%) to ecosystem respiration (ER) differences between the two half thermal years. The main environmental factors contributing to the hysteresis responses of NEE and GPP were daily accumulated radiation. Soil water content (SWC) also contributed to the hysteresis response of GPP but only at some sites. Shorter day length, lower light intensity, lower SWC and reduced photosynthetic capacity may all have contributed to the depressed GPP and net carbon uptake during the decreasing temperature seasons. The resultant hysteresis loop is an important indicator of the existence of limiting factors. As such, the role of radiation, LAI and SWC should be considered when modeling the dynamics of carbon cycling in response to temperature change.