凝胶层析在发酵液中分离提纯透明质酸中的应用

采用葡聚糖凝胶G-100,以0.1 mol/L氯化钠溶液为洗脱剂,在优化的凝胶层析条件(层析柱高度30 cm、进样量2 ml、洗脱流速1 ml/4 min)下,对透明质酸发酵液进行分离纯化,得到了高纯度的透明质酸.HA提取率达到79.85%,蛋白质去除率为84.93%.     

异育银鲫幼鱼对饲料中缬氨酸需求量的研究

通过65d的生长实验, 探讨饲料中不同缬氨酸水平对异育银鲫(Carassius auratus gibelio)幼鱼平均体重(3.17±0.01)g/尾生长性能、饲料利用和蛋白利用的影响, 确定异育银鲫对饲料中缬氨酸的需要量。实验设计由酪蛋白、明胶和晶体氨基酸为蛋白源的8种等氮(蛋白含量:38%)等能(总能:17.0 kJ/g)的半精制饲料, 其缬氨酸水平等梯度逐步升高(0.54、0.79、1.04、1.29、1.54、1.79、2.04、2.29%饲料干物质)。各实验饲料中缬氨酸的实测值分别为:0.66、0.85、1.07、1.22、1.46、1.70、1.86和2.12%饲料干物质。饲料中其他必需氨基酸含量参照异育银鲫肌肉氨基酸模式。每种饲料饲喂3个重复,每个重复30尾鱼,养殖于塑料方箱中(70 cm×55 cm×50 cm;水体容积:150 L), 养殖系统为室内流水系统(500 mL/min流速), 每天两次饱食连续投喂(9:00和16:00)。实验结果表明,饲料中适量的缬氨酸显著提高了异育银鲫的特定生长率、饲料转化效率和蛋白沉积率(P0.05), 但显著影响了血清谷氨酰胺的浓度(P0.05), 但不同的缬氨酸水平显著影响了鱼体灰分含量(P<0.05)。对各处理组异育银鲫特定生长率的结果进行二次回归分析, 结果表明满足异育银鲫最大生长的缬氨酸需求量为饲料干物质的1.72%, 即占饲料蛋白的4.53%。       

Trichogramma parasitoids for control of Lepidopteran borers in Taiwan: species,life‐history traits and Wolbachia infections

Trichogramma, polyphagous endoparasitoids of lepidopteran eggs, are used against a variety of crop pests throughout the world including those of sugar cane and corn in south‐eastern Asia. Their ability to be easily and economically reared on factitious hosts and their wide host range have contributed to their widespread use in pest control. The overall aim of this study was to select strains for eventual release in crop areas for control of lepidopteran borer pests of sugar cane and corn. To this end, we identified common Trichogramma species emerging from corn borer egg masses throughout south‐western Taiwan, compared their life‐history characteristics, assessed their thermal limits and identified the Wolbachia infection status of collected Trichogramma parasitoids. Trichogramma ostriniae was the most commonly collected species on corn, with occasional detection of T. chilonis and an unidentified species designated as T. sp. y. Although the sex ratio varied widely between sites, Wolbachia infection was detected only at a single site in one species (T. ostriniae). Wolbachia‐infected T. ostriniae were tolerant to high temperature stress. Trichogramma chilonis had lowest fecundity of the three species tested, and a Wolbachia‐infected T. ostriniae strain had lower fecundity than an uninfected strain. Given the limited availability of distribution and historical data for Trichogramma species in Taiwan, the current study provides a baseline for future work and also highlights the importance of accurately identifying species when establishing colonies of natural enemies for biocontrol.     

AEG‐1 induces gastric cancer metastasis by upregulation of eIF4E expression

Gastric cancer is the third leading cause of cancer‐related deaths worldwide, and patients with lymph node, peritoneal and distant metastasis have a poor prognosis. Overexpression of Astrocyte‐elevated gene‐1 (AEG‐1) has been reported to be correlated with the progression and metastasis of gastric cancer. However, its mechanisms are quite unclear. In this study, we found that elevated expression of AEG‐1 was correlated with metastasis in human gastric cancer tissues. Moreover, gain‐ or loss‐of‐function of AEG‐1, respectively, promoted or suppressed epithelial–mesenchymal transition (EMT), migration and invasion of gastric cancer cells. AEG‐1 positively regulated eIF4E, MMP‐9 and Twist expression. Manipulating eIF4E expression by transfection of overexpression constructs or siRNAs partially eliminated AEG‐1‐regulated EMT, cell migration and invasion. In addition, overexpression or knockdown of eIF4E promoted or suppressed EMT, cell migration and invasion in parallel with upregulation of MMP‐9 and Twist expression, while manipulating eIF4E expression partially abrogated AEG‐1‐induced MMP‐9 and Twist. Finally, silencing of AEG‐1 expression not only inhibited tumour growth in parallel with downregulation of eIF4E, MMP‐9 and Twist expression in a xenograft nude mouse model, but also suppressed lymph node and peritoneal metastasis of gastric cancer in an orthotopic nude mouse model. These findings suggest that AEG‐1 promotes gastric cancer metastasis through upregulation of eIF4E‐mediated MMP‐9 and Twist, which provides new diagnostic markers and therapeutic targets for cancer metastasis.     

Beta- Lactam Antibiotics Stimulate Biofilm Formation in Non-Typeable Haemophilus influenzae by Up-Regulating Carbohydrate Metabolism

Non-typeable Haemophilus influenzae (NTHi) is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth) stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10 µg/mL) of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended.     

Robo4 maintains vessel integrity and inhibits angiogenesis by interacting with UNC5B

Robo4 is an endothelial cell-specific member of the Roundabout axon guidance receptor family. To identify Robo4 binding partners, we performed a protein-protein interaction screen with the Robo4 extracellular domain. We find that Robo4 specifically binds to UNC5B, a vascular Netrin receptor, revealing unexpected interactions between two endothelial guidance receptors. We show that Robo4 maintains vessel integrity by activating UNC5B, which inhibits signaling downstream of vascular endothelial growth factor (VEGF). Function-blocking monoclonal antibodies against Robo4 and UNC5B increase angiogenesis and disrupt vessel integrity. Soluble Robo4 protein inhibits VEGF-induced vessel permeability and rescues barrier defects in Robo4(-/-) mice, but not in mice treated with anti-UNC5B. Thus, Robo4-UNC5B signaling maintains vascular integrity by counteracting VEGF signaling in endothelial cells, identifying a novel function of guidance receptor interactions in the vasculature.     

球孢白僵菌高渗适应性相关基因Bbmpd的克隆与表达分析

【目的】克隆与球孢白僵菌(Beauveria bassiana)的高渗适应性相关基因,并对其功能进行分析,以揭示球孢白僵菌对高渗等逆境适应的分子机理。【方法】利用YADE法克隆T-DNA的侧翼序列并进行基因组步行,获得突变基因的全长及上游序列;利用RT-PCR技术分析突变基因的表达特性以及与Bbhog1的关系;采用同源重组技术敲除Bbmpd基因。【结果】克隆得到插入突变基因及其上、下游序列全长3037bp。该基因与编码球孢白僵菌的1-磷酸甘露醇脱氢酶基因相似性为98%。Bbmpd的表达受高渗环境(0.8mol/L NaCl)的诱导,受Bbhog1信号途径的激活调节,Bbhog1缺失导致Bbmpd表达下调。Bbmpd缺失突变体在高渗胁迫下的生长受到明显抑制。Bbmpd缺失不影响球孢白僵菌在查氏培养基上的生长和产孢。【结论】由T-DNA突变体克隆了编码球孢白僵菌1-磷酸甘露醇脱氢酶基因Bbmpd,该基因的表达受高渗环境的诱导和Bbhog1的调控,与球孢白僵菌高渗适应性相关。     

小肠平滑肌胰岛素受体的特征及其蛋白激酶活性

 本实验对狗小肠平滑肌中胰岛素受体的结构和特征进行了分析研究。通过麦胚凝集素琼脂糖和两次Sepharose-CL-6B凝胶层析从平滑肌中纯化胰岛素受体,达到电泳纯。SDS-聚丙烯酰胺凝胶电泳证明胰岛素受体是由两个亚基组成的,分子量分别为135kD和90kD。磷酸化实验证明平滑肌胰岛素受体具有胰岛素依赖性蛋白激酶活性,能催化自身的β亚基磷酸化和底物的磷酸化。Scatchard分析表明胰岛素和受体的结合呈(?)协同效应,最大结合率为13μg胰岛素/mg蛋白质。     

黄花菜的采后生理与保鲜技术研究进展

黄花菜(Hemerocallis citrina Baroni,daylily)在我国有几千年的种植历史,是一种具有较高经济价值的特色蔬菜。通过对国、内外在黄花菜保鲜和贮藏的研究的了解,从黄花菜的生理、保鲜两方面对黄花菜的采后生理及保鲜技术进行了综述,同时也对黄花菜的应用价值进行了介绍。