中国大鲵研究进展

中国大鲵是我国特有濒危的两栖物种,是研究生物进化、生物多样性、性别决定分子机制等问题的好材料。近年来,人们对它的研究力度不断加大,著述颇多。本文对大鲵的生态保护、解剖发育、生理生化、遗传进化等方面的近期研究资料进行了整理和回顾,也简要探讨了今后大鲵研究的主要工作,以期为研究者们提供有价值的资料。     

The Arabidopsis RING E3 ubiquitin ligase AtAIRP2 plays combinatory roles with AtAIRP1 in abscisic acid-mediated drought stress responses

The ubiquitin (Ub)-26S proteasome pathway is implicated in various cellular processes in higher plants. AtAIRP1, a C3H2C3-type RING (for Really Interesting New Gene) E3 Ub ligase, is a positive regulator in the Arabidopsis (Arabidopsis thaliana) abscisic acid (ABA)-dependent drought response. Here, the AtAIRP2 (for Arabidopsis ABA-insensitive RING protein 2) gene was identified and characterized. AtAIRP2 encodes a cytosolic C3HC4-type RING E3 Ub ligase whose expression was markedly induced by ABA and dehydration stress. Thus, AtAIRP2 belongs to a different RING subclass than AtAIRP1 with a limited sequence identity. AtAIRP2-overexpressing transgenic (35S:AtAIRP2-sGFP) and atairp2 loss-of-function mutant plants exhibited hypersensitive and hyposensitive phenotypes, respectively, to ABA in terms of seed germination, root growth, and stomatal movement. 35S:AtAIRP2-sGFP plants were highly tolerant to severe drought stress, and atairp2 alleles were more susceptible to water stress than were wild-type plants. Higher levels of drought-induced hydrogen peroxide production were detected in 35S:AtAIRP2-sGFP as compared with atairp2 plants. ABA-inducible drought-related genes were up-regulated in 35S:AtAIRP2-sGFP and down-regulated in atairp2 progeny. The positive effects of AtAIRP2 on ABA-induced stress genes were dependent on SNF1-related protein kinases, key components of the ABA signaling pathway. Therefore, AtAIRP2 is involved in positive regulation of ABA-dependent drought stress responses. To address the functional relationship between AtAIRP1 and AtAIRP2, FLAG-AtAIRP1 and AtAIRP2-sGFP genes were ectopically expressed in atairp2-2 and atairp1 plants, respectively. Constitutive expression of FLAG-AtAIRP1 and AtAIRP2-sGFP in atairp2-2 and atairp1 plants, respectively, reciprocally rescued the loss-of-function ABA-insensitive phenotypes during germination. Additionally, atairp1/35S:AtAIRP2-sGFP and atairp2-2/35S:FLAG-AtAIRP1 complementation lines were more tolerant to dehydration stress relative to atairp1 and atairp2-2 single knockout plants. Overall, these results suggest that AtAIRP2 plays combinatory roles with AtAIRP1 in Arabidopsis ABA-mediated drought stress responses.     

Rapid determination of gatifloxacin in biological samples and pharmaceutical products using europium‐sensitized fluorescence spectrophotometry

A europium‐sensitized fluorescence spectrophotometry method using an anionic surfactant, sodium dodecyl benzene sulphonate (SDBS), was developed for the determination of gatifloxacin (GFLX). The GFLX–Eu³⁺–SDBS system was studied and it was found that SDBS significantly enhanced the fluorescence intensity of the GFLX–Eu³⁺ complex (about 25‐fold). The optimal experimental conditions were determined as follows: excitation and emission wavelengths of 338 and 617 nm, pH 7.5, 3.0 × 10^–6 mol/L europium(III), and 5.0 × 10^–5 mol/L SDBS. The enhanced fluorescence intensity of the system (ΔI_f) showed a good linear relationship with the concentration of GFLX over the range 1.0 × 10^–8–8.0 × 10^–7 mol/L with a correlation coefficient of 0.9990. The detection limit (S:N = 3) was determined as 1.0 × 10^–9 mol/L. This method has been successfully applied for the determination of GFLX in pharmaceuticals and human urine/serum samples. Compared with most other methods reported, the rapid and simple procedure proposed here offered higher sensitivity, wider linear range and good stability. The luminescence mechanism of the system is also discussed in detail. Copyright © 2008 John Wiley & Sons, Ltd.     

Lipolysis drives expression of the constitutively active receptor GPR3 to induce adipose thermogenesis

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饲料中灵芝提取物对异育银鲫生长、饲料利用、免疫应答及抗病的影响

通过生长实验探求饲料中添加灵芝提取物饲料对异育银鲫（6.720.11） g生长、饲料利用及免疫应答的影响,以及停止灵芝提取物饲料后,使用效果的持续性。在实验进行的第21天（D21）、第46天（D46）、第65天（D65）进行取样,结果表明所有的灵芝提取物组的特定生长率在饲喂65d后与对照组无显著差异（P0.05）。摄食率随着灵芝提取物在饲料中含量的增加而增加（P0.05）,饲料效率则有着相反的趋势（P0.05）。在D46和D65时,随着灵芝提取物的含量增高,吞噬活力略有增高,但无显著差异（P0.05）。在D21、D46和D65时,呼吸暴发活力随着灵芝提取物的含量增高而增高（P0.05）,在D21时,（0.6%-21d、0%-44d）处理组和（3%-21d、0%-44d）处理组其呼吸暴发活力高于对照组（P0.05）。在D65时,（0.6%-21d、0%-44d）处理组和（3%-21d、0%-44d）处理组其呼吸暴发活力仍高于对照组（P0.05）,但差异不显著。D21、D46和D65时,替代途径补体溶血活力随着灵芝提取物的增高而降低（P0.05）。所有处理组溶菌酶活力在D21时没有产生显著差异。在D46时,对照组溶菌酶活力比最高添加组低但高于低添加组（P0.05）,（3%-1d、0%-44d）处理组也仍高于对照组（P0.05）。在D65时,所有添加组的溶菌酶活力均低于对照组（P0.05）。在D65时,髓过氧化物酶活力随着灵芝提取物的添加剂量升高而升高（P0.05）,（0.6%-21d、0%-44d）处理组仍高于对照组（P0.05）。灵芝提取物的能提高白细胞呼吸暴发活力,且在在高剂量摄入21d后（D46时）,仍可以保持较高的活力,说明灵芝提取物对免疫应答有一定时间延续效应。随着灵芝提取物添加量的升高,异育银鲫在经爱德华氏菌攻毒后的存活率显著提高。最高存活率组为3%添加组,显著高于对照组（P0.05）。实验周期为65d时,饲料中灵芝提取物推荐添加剂量为3%。       

木本植物内源生长素免疫胶体金定位分析技术的研究

以木本植物杨树(Populus sp.)和核桃(Juglans regia L.)为材料,对内源生长素免疫胶体金定位技术在固定、烤片、免疫染色、显色等关键环节进行了改进优化与验证.结果显示,优化后适合木本植物定位方法的主要技术要点是:在染色中,通过采用尿素-胰蛋白酶联合消化技术和增加牛血清白蛋白处理大大地改善了抗原修复结果,提高了染色的敏感性和特异性.利用优化后的方法对核桃幼胚和杨树嫩茎诱导生根过程中的吲哚乙酸(IAA)进行定位研究,发现杨树试管嫩茎生根过程中,形成层及周缘维管束有很强的IAA信号,核桃子叶生根中,胚中有很强的IAA信号,胚根中有半圆形、胚芽中有>"形强信号区,胚轴信号较弱,胚根信号最强.研究表明,与传统免疫染色方法相比,优化后的方法对木本植物生长素定位具有敏感性高,特异性强,银颗粒明显,背景清晰,耗时少等特点.     

几个生态因子对黄盖鲽的影响

黄盖鲽(Psuedopleuronectes yokohameGünther)是鲽科中的冷温性鱼类,广泛分布在日本、朝鲜及我国的东海、黄海和渤海。地方种群多,洄游范尉小,适应能力强,既是底栖生物食性,又可耐较低的温度,虽非名贵鱼种,却具有一定的经济价值,在开展近海增殖时,不存在越冬和效益外流问题。因此,早在60年代,日本山口县和大分县的学者们,便开始着手该品种的人工孵化和增殖放流试验。80年代初已达年放流2—3cm的稚鱼苗7.0×10~6尾的水平。为了在渤海开展增殖工作,建立综     

Substrate modification with lysine 63-linked ubiquitin chains through the UBC13-UEV1A ubiquitin-conjugating enzyme

Protein modification with lysine 63-linked ubiquitin chains has been implicated in the non-proteolytic regulation of signaling pathways. To understand the molecular mechanisms underlying this process, we have developed an in vitro system to examine the activity of the ubiquitin-conjugating enzyme UBC13-UEV1A with TRAF6 in which TRAF6 serves as both a ubiquitin ligase and substrate for modification. Although TRAF6 potently stimulates the activity of UBC13-UEV1A to synthesize ubiquitin chains, it is not appreciably ubiquitinated. We have determined that the presentation of Lys(63) of ubiquitin by UEV1A suppresses TRAF6 modification. Based on our observations, we propose that the modification of proteins with Lys(63)-linked ubiquitin chains occurs through a UEV1A-independent substrate modification and UEV1A-dependent Lys(63)-linked ubiquitin chain synthesis mechanism.     

Simulation of sequential batch reactor (SBR) operation for simultaneous removal of nitrogen and phosphorus

Modeling of the operation of sequential batch reactor (SBR) was performed to find out optimum design parameters for simultaneous removal of nitrogen and phosphorus in a small-scale wastewater treatment plant. The models were set up with material balances on SBR operation and Monod kinetics. The model parameters were obtained to best fit the experimental results in a small scale SBR. The models were useful in optimizing hydraulic retention time (HRT) and successfully simulated operations of SBR in a larger scale. Especially the model predicted well the reactions occurring in the filling period as well as the effect of dilution, and evaluated the performance of SBR process under diverse operating conditions.     

MST1 promotes apoptosis through phosphorylation of histone H2AX

MST1 (mammalian STE20-like kinase 1) is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. Overexpression of MST1 induces apoptotic morphological changes such as chromatin condensation, but the mechanism is not clear. Here we show that MST1 induces apoptotic chromatin condensation through its phosphorylation of histone H2AX at Ser-139. During etoposide-induced apoptosis in Jurkat cells, the cleavage of MST1 directly corresponded with strong H2AX phosphorylation. In vitro kinase assay results showed that MST1 strongly phosphorylates histone H2AX. Western blot and kinase assay results with a mutant S139A H2AX confirmed that MST1 phosphorylates H2AX at Ser-139. Direct binding of MST1 and H2AX can be detected when co-expressed in HEK293 cells and was also confirmed by an endogenous immunoprecipitation study. When overexpressed in HeLa cells, both the MST1 full-length protein and the MST1 kinase domain (MST1-NT), but not the kinase-negative mutant (MST1-NT-KN), could induce obvious endogenous histone H2AX phosphorylation. The caspase-3 inhibitor benzyloxycarbonyl-DEVD-fluoromethyl ketone (Z-DEVD-fmk) attenuates phosphorylation of H2AX by MST1 but cannot inhibit MST1-NT-induced histone H2AX phosphorylation, indicating that cleaved MST1 is responsible for H2AX phosphorylation during apoptosis. Histone H2AX phosphorylation and DNA fragmentation were suppressed in MST1 knockdown Jurkat cells after etoposide treatment. Taken together, our data indicated that H2AX is a substrate of MST1, which functions to induce apoptotic chromatin condensation and DNA fragmentation.