Advanced glycosylation end products promote migration of podocytes through mTORC1/uPAR pathway

AIM: To investigate the influence of advanced glycosylation end products-modified bovine serum albumin (AGE-BSA) on mammalian target of rapamycin complex 1 (mTORC1), urokinase-type plasminogen activator receptor (uPAR), and cell mobility in the podocytes, and to further explore the probable relationship. METHODS: The conditionally immortalized mouse podocyte cell line was cultured in vitro. MTT assay and immunofluorescence were used to analyze the cell viability and cytoskeleton of the podocytes treated with the stimuli and intervention agents. The activity of mTORC1 and the expression level of uPAR in normal podocytes and podocytes treated with control BSA or AGE-BSA were detected by Western blotting. The migration ability of the podocytes was determined by would-healing assay. Rapamycin was added to inhibit the activity of mTORC1 along with the addition of AGE-BSA to observe the changes of uPAR and the motility of podocytes. RESULTS: No significant difference of the cell viability or cytoskeleton in the podocytes treated with the stimuli and intervention agents was observed. AGE-BSA up-regulated the activity of mTORC1 and the expression of uPAR, and induced the high mobility of the podocytes. Rapamycin obviously reduced the high expression level of uPAR and the increase in the migration ability of podocytes caused by AGE-BSA treatment. CONCLUSION: AGE-BSA might cause the high migration of podocytes through the mTORC1/uPAR signaling pathway.     

多酶激活剂对小白菜生长、品质及抗氧化酶活性的影响

研究了不同浓度的多酶激活剂对小白菜生长、品质及抗氧化酶活性的影响。研究结果表明,多酶激活剂处理均可提高小白菜的地上部鲜、干质量、VC和可溶性蛋白含量,降低小白菜硝酸盐含量,增强超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）活性,以T2处理（800倍液）效果最佳。     

氮气与甲酸乙酯混合熏蒸对锯谷盗的毒力研究

采用广口瓶密闭熏蒸法,在实验室条件下研究了氮气与甲酸乙酯混合熏蒸对锯谷盗成虫的毒力。结果表明,氮气和甲酸乙酯混用具有良好的熏蒸效果。在81%、87%和95%的氮气浓度下,甲酸乙酯为10μL/L熏蒸处理24h,锯谷盗的校正死亡率分别为55%、75%和97%,表明氮气可显著提高甲酸乙酯的熏蒸毒力;温度对氮气和甲酸乙酯混合熏蒸效果具有显著影响作用,相同条件下,在30℃时校正死亡率为41%,在15℃时则提高至90%,表明低温条件下熏蒸效果比高温条件下更好。     

黑曲霉果胶裂解酶基因的克隆与原核表达

根据NCBI上果胶裂解酶(PL)基因在黑曲霉基因组上的分布特点设计特异引物,通过重组PCR扩增出黑曲霉编码PL的结构基因。将PL基因连接到大肠杆菌表达载体pET-28a上,得到重组质粒pET-28a-PL,转化大肠杆菌(E.coli)BL21后,用IPTG进行诱导表达。结果表明,PL基因片段序列全长1 431bp,与已知PL基因(XM_001397206.2)核苷酸和氨基酸序列同源性均为99%。表达产物经12%SDS-PAGE电泳,得到了一条大小约为50kD的条带,与预期分子量相符,证明真菌PL基因在E.coli BL21中可以有效表达。     

Characterization of a New Biofunctional,Exolytic Alginate Lyase from Tamlana sp. s12 with High Catalytic Activity and Cold-Adapted Features

Alginate, a major acidic polysaccharide in brown algae, has attracted great attention as a promising carbon source for biorefinery systems. Alginate lyases, especially exo-type alginate lyase, play a critical role in the biorefinery process. Although a large number of alginate lyases have been characterized, few can efficiently degrade alginate comprised of mannuronate (M) and guluronate (G) at low temperatures by means of an exolytic mode. In this study, the gene of a new exo-alginate lyase—Alys1—with high activity (1350 U/mg) was cloned from a marine strain, Tamlana sp. s12. When sodium alginate was used as a substrate, the recombinant enzyme showed optimal activity at 35 °C and pH 7.0–8.0. Noticeably, recombinant Alys1 was unstable at temperatures above 30 °C and had a low melting temperature of 56.0 °C. SDS and EDTA significantly inhibit its activity. These data indicate that Alys1 is a cold-adapted enzyme. Moreover, the enzyme can depolymerize alginates polyM and polyG, and produce a monosaccharide as the minimal alginate oligosaccharide. Primary substrate preference tests and identification of the final oligosaccharide products demonstrated that Alys1 is a bifunctional alginate lyase and prefers M to G. These properties make Alys1 a valuable candidate in both basic research and industrial applications.     

不同烤烟品种叶片代谢关键酶活性差异研究

以翠碧一号、K326、中烟100、云烟87、红花大金元、中烟103为试验材料,通过大田试验研究了不同烤烟品种叶片不同生育时期代谢关键酶活性以及质量差异。结果表明:从栽后25d开始,烟叶谷氨酰胺合成酶(GS)活性呈双峰曲线,K326和云烟87最低,翠碧一号和中烟103前高后低;淀粉酶活性随烟叶生长而增长并到55d时出现峰值,随后开始下降,以翠碧一号、云烟87的淀粉酶活性最高;苯丙氨酸解氨酶(PAL)活性在栽后65d之前,中烟100和中烟103显著的低于其他4个品种;多酚氧化酶(PPO)活性在栽后45d品种间开始出现差异,以翠碧一号活性最低。从烤后烟样化学成分含量来看,中烟103的总氮、烟碱含量最低,还原糖、总糖含量最高,钾氯含量适中,化学成分比较协调。     

一般氮代谢调控蛋白NtrC的研究进展

NtrBC是细菌氮代谢调控系统中双组分系统,其中NtrC是响应环境信号的转录激活蛋白,对靶基因具有转录激活作用。已有的研究表明NtrC在细菌氮源利用、生物固氮、高分子胞外聚合物合成、碳氮平衡等方面发挥了重要的调节作用。对NtrC的全局调控作用研究,是当前微生物代谢调控网络研究热点之一。综述了细菌NtrC近年来的研究进展,详细介绍了细菌NtrC的生物学功能及调控机制,以期为更好地了解微生物的环境适应性,代谢多样性和碳氮偶联作用的分子机制奠定理论基础。     

Short-Term Storage of Semen of Rainbow Trout: Interactions of Time,Antibiotic, and Activator

This study was designed to examine the main effects and interactions of time, presence of antibiotics, and type of sperm activators on the fertilization capacity (eyeing rate) of refrigerated semen of rainbow trout, Oncorhynchus mykiss. The semen samples were stored in the presence or absence of 250 IU ml^?1 penicillin and 250 μg ml^?1 streptomycin sulfate. Freshwater and a saline solution were used as sperm activators. The semen samples were stored at 2–3°C and fertilized after 0, 6, 8, 12, 19, and 25 days of storage. Fertilizing capacities of semen samples stored in the presence of antibiotics (63.8 ± 5.6%) were significantly (p < 0.05) higher than those stored in the absence of antibiotics (46.2 ± 6.7%). Also, the fertilizing capacities of stored semen samples activated using saline solution (70.7 ± 5.7%) had significantly (p < 0.05) higher values than those activated using freshwater (39.3 ± 5.9%). Semen samples stored in the absence of antibiotics completely lost fertilizing capacity within 19 days of storage. After 25 days of storage in the presence of antibiotics, induction of fertilization using freshwater and saline solution resulted in 0% and 79.8 ± 1.7% fertility, respectively.     

绿叶挥发物代谢调控及分子机理研究进展

植物处于复杂的自然环境中,常受到植食性昆虫、病原菌及机械损伤等生物或非生物的侵害并造成植物的局部伤害,从而启动复杂的防御反应。释放绿叶挥发物（green leaf volatiles,GLVs）是植物防御性反应之一。绿叶挥发物是植物不饱和脂肪酸经过Oxylinpins生化途径中的脂氧合酶（LOX ）和脂氢过氧化物裂解酶（HPL）催化而形成的一类C6和C9醛、醇及其相应的酯类。作为启动植物防御机制的信号分子,这类挥发性物质通过长距离的气态传输在植物与病原菌、植物与植食性昆虫、植物自身和相邻的植物之间起作用,从而介导防御性反应。研究其生化途径及其调节机理,对探索其对病虫害的直接、间接防御、改善作物风味品质及抗病新种质创制起着非常重要的作用,且对园艺植物甚至农林生态系统中作物病虫害综合治理策略的制定具有重要的指导意义。该文综述了绿叶挥发物合成的生化途径及其作为信号转导物质的作用机制,并阐述了绿叶挥发物在植物抗逆方面的生理作用及其应用前景。     

尾叶桉不同无性系抗病性与酶活性关系的研究

应用菌液培养法和分生孢子悬浮液喷雾法直接测定尾叶桉无性系对青枯病和焦枯病的抗病能力,4个无性系的抗病能力排序为 GUL4＞GUL1=GUL8＞GUL15.对这4个无性系进行过氧化物酶(POD)活性测定、同工酶电泳分析和苯丙氨酸解氨酶(PAL)活性测定,结果表明抗病性越强的无性系,其过氧化物酶和苯丙氨酸解氨酶的活性越高,过氧化物酶同工酶谱带数量越多.