容积调控氯通道的研究进展

细胞在低渗环境中出现渗透性膨胀 ,随后细胞内的溶质及水分外流 ,使已膨胀的细胞容积向正常容积转化 ,此过程称为调节性细胞容积减小 (RVD) ,它是哺乳动物细胞普遍存在的现象。容积调控氯通道 (VRAC)在这个过程中起重要作用 ,不仅如此 ,最近研究发现VRAC参与了细胞增殖、分化和凋亡过程。本文主要综述了VRAC的生物学特点和生理功能等方面的研究进展     

砷对蓝藻光合作用和细胞生长的影响

在世界的许多地方, 砷污染是一个严重的问题, 尤其是水体中的污染程度更加严重。砷主要以As( V) 和As( ó ) 形式存在。由于AsO43- 在结构上与PO43-很相似, 它通过磷转运途径进入细胞, 因此砷的毒性主要是源于对磷代谢的干扰1 。砷对于人和动物的危害性是人所共知的, 它会引发细胞凋亡, 导致多种组织和器官发生癌变, 从而引起死亡。但是砷对于水体生态系统中的重要类群) 藻类的生理和生长的影响报道较少。       

Site-selective lateral multilayer assembly of bienzyme with polyelectrolyte on ITO electrode based on electric field-induced directly layer-by-layer deposition

We describe here a new approach to construct a multilayer enzyme/polyelectrolyte film on a structured transparent indium-tin oxide (ITO) covered glass electrode surface as micropattern, on which two different types of enzyme distributed laterally on one common substrate without interference. The multilayer film was prepared by alternate electric field directed layer-by-layer assembly deposition and alternate deposition of different redox enzymes and polyelectrolyte poly(diallyldimethylammonium chloride) (PDDA) onto the site-selective ITO glass electrode surface. The cyclic voltammogram, obtained from the ITO glass electrode modified with the glucose oxidase (GO(X))/PDDA and catalase (CA(T))/PDDA multilayers, revealed that the bioelectrocatalytic response is directly correlated to the number of deposition bilayers. From the analysis of cyclic voltammetric characterization, the coverage of catalytically active enzymes per enzyme/PDDA bilayer during the multilayer formation was homogeneous, which demonstrates that the multilayer is constructed in a spatially ordered manner. Also, from the atomic force microscopy and Brewster angle microscopy measurements, more information of the multilayer constructed by different methods on the modified electrode surface is obtained and compared. This fabrication technique is simple and would be applicable to the construction of a thickness- and area-controlled biopattern composed of multi-enzymes as well as multiple biomaterials.     

拉布拉多犬的母性行为水平对幼犬胆量的影响

母性行为是动物以维持幼崽的生存及生理健康为主要目的的一种基本行为,母性行为作为重要的早期经历对动物的个体发展有深远影响。动物的行为在时间和环境中具有一致性,多个行为特征的一致性加权被称为气质特征,气质特征的差异是犬(Canis lupus familiaris)能否顺利通过培训成为导盲犬的决定性因素。其中,胆量是决定导盲犬培训成功与否的重要气质特征。本研究以中国导盲犬大连培训基地的拉布拉多种犬及幼犬为研究对象,探究母性行为水平对幼犬胆量的影响。本研究通过视频观察记录拉布拉多犬哺乳期前21 d的母性行为变量时长,对在哺乳区内、身体接触、哺乳和舔舐幼犬4项变量进行主成分分析后将7只实验犬分为母性行为高水平与低水平两组。对两组犬生产的共54只幼犬于6 ~ 8周龄时进行幼犬胆量行为测试,根据胆量行为测试的评分标准对幼犬的行为表现进行评分,统计分析母性行为高水平组与低水平组其幼犬的胆量是否存在差异。本研究结果表明,母性行为低水平组的幼犬在胆量测试中面对陌生环境、突然出现的响声刺激、突然打开的雨伞刺激以及陌生人的游戏邀请时均表现出更大的胆量。在被动测试中,母性行为低水平组幼犬的探索潜伏时长显著短于母性行为高水平组幼犬(P < 0.05),探索范围显著大于母性行为高水平组幼犬(P < 0.05),紧张程度极显著低于母性行为高水平组幼犬(P < 0.01);在金属响声测试中,母性行为低水平组幼犬的惊吓反应(P < 0.01)和紧张程度(P < 0.01)均极显著低于母性行为高水平组幼犬;在雨伞测试中,母性行为低水平组幼犬的紧张程度显著低于母性行为高水平组幼犬(P < 0.05);在玩具测试中,母性行为低水平组幼犬的玩耍兴趣显著高于母性行为高水平组幼犬(P < 0.05),紧张程度显著低于母性行为高水平组幼犬(P < 0.05);在斜坡隧道测试中,母性行为低水平组幼犬的紧张程度显著低于母性行为高水平组幼犬(P < 0.05),通过斜坡的用时短于母性行为高水平组幼犬,但经统计检验无显著差异(P > 0.05)。本研究的结论为低母性行为水平带给幼犬强度适当的早期生活压力,使幼犬面对新环境刺激时表现出更好的适应能力和较大的胆量。本研究为工作犬种犬的筛选提出新的建议：母性行为水平低的种犬对幼犬胆量的发展有更好的影响。     

中国东、南部的3个腔蚓属蚯蚓新物种

本文报道了寡毛纲(Oligochaeta)巨蚓科(Megascolecidae)腔蚓属(Metaphire)3个新发现物种,分别是象头山腔蚓(M. xiangtoumontis Dong & Jiang sp. nov.),韩摆渡腔蚓(M. hanbaiduensis Dong & Sun sp. nov.)和长白山腔蚓(M. changbaimontis Dong & Shen sp. nov.)。象头山腔蚓受精囊孔2对,位于7/8 ~ 8/9节间,属于M. insulana物种群。韩摆渡腔蚓受精囊孔3对,位于6/7 ~ 8/9节间,属于M. houlleti物种群。长白山腔蚓受精囊孔2对,位于6/7 ~ 7/8节间,属于M. glandularis物种群。所有新物种均附有形态学描述、图片、与相似物种的形态学比较及与GenBank上亲缘关系相近物种的遗传距离计算分析。以上结果丰富了我国腔蚓属蚯蚓的物种多样性,并首次记录了采集于长白山国家级自然保护区的巨蚓科蚯蚓新物种。     

Detection and Quantification of Fusarium oxysporum f. sp. niveum Race 1 in Plants and Soil by Real-time PCR

Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is the most serious soil-borne disease in the world and has become the main limiting factor of watermelon production. Reliable and quick detection and quantification of Fon are essential in the early stages of infection for control of watermelon Fusarium wilt. Traditional detection and identification tests are laborious and cannot efficiently quantify Fon isolates. In this work, a real-time polymerase chain reaction (PCR) assay has been described to accurately identify and quantify Fon in watermelon plants and soil. The FONRT-18 specific primer set which was designed based on identified specific sequence amplified a specific 172 bp band from Fon and no amplification from the other formae speciales of Fusarium oxysporum tested. The detection limits with primers were 1.26 pg/μl genomic DNA of Fon, 0.2 pg/ng total plant DNA in inoculated plant, and 50 conidia/g soil. The PCR assay could also evaluate the relationships between the disease index and Fon DNA quantity in watermelon plants and soil. The assay was further used to estimate the Fon content in soil after disinfection with CaCN₂. The real-time PCR method is rapid, accurate and reliable for monitoring and quantification analysis of Fon in watermelon plants and soil. It can be applied to the study of disease diagnosis, plant-pathogen interactions, and effective management.     

Mitotic checkpoint gene expression is tuned by codon usage bias

The mitotic checkpoint (also called spindle assembly checkpoint, SAC) is a signaling pathway that safeguards proper chromosome segregation. Correct functioning of the SAC depends on adequate protein concentrations and appropriate stoichiometries between SAC proteins. Yet very little is known about the regulation of SAC gene expression. Here, we show in the fission yeast Schizosaccharomyces pombe that a combination of short mRNA half‐lives and long protein half‐lives supports stable SAC protein levels. For the SAC genes mad2 ⁺ and mad3 ⁺, their short mRNA half‐lives are caused, in part, by a high frequency of nonoptimal codons. In contrast, mad1 ⁺ mRNA has a short half‐life despite a higher frequency of optimal codons, and despite the lack of known RNA‐destabilizing motifs. Hence, different SAC genes employ different strategies of expression. We further show that Mad1 homodimers form co‐translationally, which may necessitate a certain codon usage pattern. Taken together, we propose that the codon usage of SAC genes is fine‐tuned to ensure proper SAC function. Our work shines light on gene expression features that promote spindle assembly checkpoint function and suggests that synonymous mutations may weaken the checkpoint.     

A sensitive signal-on assay for MTase activity based on methylation-responsive hairpin-capture DNA probe

This work develops a simple, sensitive and signal-on electrochemical sensor for methyltransferase (MTase) activity analysis. The sensor is composed of a methylene blue-modi?ed "signaling DNA probe" and a "capture DNA probe" tethered methylation-responsive hairpin DNA (hairpin-capture DNA probe). The thiol- modified hairpin-capture DNA probe at 5' end was firstly self-assembled on gold electrode via Au-S bonding. Methylation-induced scission of hairpin-capture DNA probe would displace the hairpin section and remain the "capture DNA probe" section on the gold electrode. Subsequently, the remained "capture DNA probe" on the gold electrode can hybridize with the methylene blue-modi?ed "signaling DNA probe", mediating methylene blue onto the gold electrode surface to generate redox current. It was eT on state. The developed facile signal-on electrochemical sensing system showed a linear response to concentration of Dam MTase range from 0.1 to 1.0 U/mL. The detection limit of Dam MTase activity was determined to be 0.07 U/mL and the total detection time is 7h. The sensor also has the ability to provide information about the dynamics of methylation process. Furthermore, we demonstrated that this sensor could be utilized to screen inhibitors or drugs for Dam MTase.     

重寄生真菌盾壳霉pH信号通路中Pal相关基因的功能鉴定

【目的】研究pH信号通路(Pal)在重寄生真菌盾壳霉与寄主核盘菌互作过程中的作用。【方法】从盾壳霉全基因组信息中分析获得了6个Pal相关基因CmpalA、CmpalB、CmpalC、CmpalF、CmpalH和CmpalI的全编码序列和氨基酸序列,通过PEG介导的原生质转化技术获得了CmpalA、CmpalB、CmpalC、CmpalF和CmpalH等5个基因的敲除突变体,分析这些敲除突变体与野生型在菌落培养性状、重寄生能力、降解草酸能力、产生抗真菌物质能力等方面的差异。【结果】与野生型相比,在pH 6–8的条件下,5个Pal相关基因敲除突变体的菌丝生长受到显著抑制,这说明缺失Pal相关基因使盾壳霉对高pH值环境更加敏感。菌核重寄生试验发现5个Pal相关基因敲除突变体的重寄生能力均显著低于野生型。qRT-PCR试验结果表明,敲除Pal相关基因之后导致重寄生相关酶基因Cmch1、Cmg1和Cmsp1的表达量显著降低,而且pH信号通路下游的CmpacC基因的表达量也显著降低。Pal相关基因敲除突变体在pH 6条件下对草酸盐的降解能力显著高于野生型,同时这5个突变体在pH 8条件下产生抗真菌物质能力也显著高于野生型。【结论】pH信号通路相关基因的缺失影响盾壳霉对环境pH的响应。pH信号通路在盾壳霉与核盘菌互作中发挥重要作用,不仅影响盾壳霉的重寄生作用,而且还影响盾壳霉的草酸降解作用和抗真菌作用。     

Spindlin1, a novel nuclear protein with a role in the transformation of NIH3T3 cells

spindlin1, a novel human gene recently isolated by our laboratory, is highly homologous to mouse spindlin gene. In this study, we cloned cDNA full-length of this novel gene and send it to GenBank database as spindlin1 (Homo sapiens spindlin1) with Accession No. AF317228. In order to investigate the function of spindlin1, we studied further the subcellular localization of Spindlin1 protein and the effects of spindlin1 overexpression in NIH3T3 cells. The results showed that the fusion protein pEGFP-N1-spindlin1 was located in the nucleus and the C-terminal is correlated with nuclear localization of Spindlin1 protein. NIH3T3 cells which could stably express spindlin1 as a result of RT-PCR analysis compared with the control cells displayed a complete morphological change; made cell growth faster; and increased the percentage of cells in G2/M and S phase. Furthermore, overexpressed spindlin1 cells formed colonies in soft agar in vitro and formed tumors in nude mice. Our findings provide direct evidence that spindlin1 gene may contribute to tumorigenesis.