L-茶氨酸对H2O2致L02细胞损伤的保护作用及其机制研究

研究L-茶氨酸对肝细胞损伤的保护作用及其机制。利用H2O2诱导的LO2肝细胞损伤模型,分别用MTT法检测细胞存活率、测定LDH、流式细胞术检测细胞凋亡率、Western blot法检测Caspase-3和PARP蛋白表达7LBax／Bcl-2比值的变化,评价L-茶氨酸是否能保护H2O2诱导的肝细胞损伤。结果表明,L-茶氨酸能提高H2O2损伤的L02细胞存活率,减少LDH的渗漏,降低肝细胞凋亡,且L-茶氨酸通过抑制caspase-3的激活和PARP的切割及Bax／Bcl-2比值的升高而发挥抗凋亡的作用。L-茶氨酸对肝细胞损伤有一定的治疗和保护作用。     

维生素C对嗜酸乳杆菌GIM1.208 β-葡萄糖苷酶激活效应及分子互作关系

【背景】研究发现维生素C(vitaminC,VC)对嗜酸乳杆菌GIM1.208β-葡萄糖苷酶(β-glucosidase, BGL)具有明显激活作用。【目的】探究VC对BGL的影响特性以及二者互作关系。【方法】通过对硝基苯酚法、电化学法、Zeta电位法及核磁共振氢谱法研究VC对BGL的活性影响及互作特性。【结果】VC对BGL催化底物活性具有明显促效作用,Km=1.167 mmol/L,Vmax随着VC浓度增大而增大;VC浓度为3.5%时酶活性达到最高;在VC浓度为3.5%、温度低于30℃时,酶活性受温度影响较小,VC对BGL具有良好的稳定作用,相对酶活保持在90%以上。VC与BGL结合,两者存在弱静电作用且两者的相互作用使得VC的电氧化反应变得更加容易,同时推测VC分子中乙二醇支链片段与BGL之间存在氢键作用。【结论】VC对BGL具有明显激活作用,试验探究了两者作用力类型及作用位点,为BGL应用及天然酶激活剂的高效利用提供设计理论参考。     

树木年轮中的微量元素含量与克山病关系的初步分析

森林是陆地生态系统中的一个重要组成部分,在一定程度上可以说它是“历史的见证”,因为在森林中记录了过去生态环境的一些变化和特点。 树木年轮是过去环境记录的一个重要各面,大家知道,通过观察树木年轮,可分析研究近千年来地球上气候的变化。这一方法在国内外已得到广泛的应用。但是从生物地球化学观     

水稻幼苗活力性状的低温反应数量性状基因座检测

以籼粳交“密阳23/吉冷1号”的F2：3代200个家系作为作图群体,在12℃冷水胁迫下,进行苗高、苗鲜重和苗干重等水稻幼苗活力性状的低温反应鉴定,并利用由SSR标记构建的分子连锁图谱为基础,对冷水胁迫下苗高、苗鲜重和苗干重以及它们的低温反应指数进行了数量性状基因座（QTLs）检测。研究结果表明,低温胁迫下上述幼苗活力性状在F3家系群中均表现为接近正态的连续分布,表现为由多基因控制的数量性状;在第1、2、7、8和12染色体上,检测到与幼苗活力性状的低温反应相关的QTL共12个,对表型变异的贡献率范围为5．2％-17．9％,其中位于第2染色体RM262-RM263区间和第12染色体RM270-RM17区间的与低温下苗高相关的qCSH2和qCSH12,以及位于第12染色体RM19-RM270区间和第1染色体RM129-RM9区间的分别控制低温下苗干重及其低温反应指数的qSDW12和qCSDW1对表型变异的贡献率较大,分别为16．6％、17．9％、15．9％和16．2％。其增效等位基因均来自吉冷1号,前两者均表现为加性效应,后两者分别表现为显性和超显性。     

分子标记技术的发展及应用

介绍了几种应用前景较广的分子标记,如基于DNA杂交技术的分子标记:限制性片段长度多态性(RFLP)和DNA可变串联重复数标记(VNRT);基于PCR技术的分子标记:随机扩增多态性 DNA(RAPD)、酶切扩增多态性(CAPS)、扩增片段长度多态性(AFLP)、微卫星DNA(SSR)和DNA单链构象多态性(SSCP);以及新兴的第3代分子标记,即基于DNA芯片技术的分子标记:单核苷酸多态性(SNP)等。分别阐述了它们的原理、方法步骤与优缺点、应用注意事项和适用范围,同时概述了它们在生物学研究中的应用和进展。     

蒙古黄芪病程相关蛋白AmPR-10于大肠杆菌不同载体的可溶性表达策略研究

蒙古黄芪病程相关蛋白（Astragalus membranaceus pathogenesis-related protein-10, AmPR-10）具有核酸酶活性,对黄芪生长发育及抗病机制具有重要意义。但从天然黄芪中提取蛋白质的传统方法成本较高,蛋白得率较少。研究首次利用大肠杆菌表达体系对AmPR-10进行可溶性外源表达,构建了3种重组子：①以pET28a为载体构建pET28a-AmPR-10;②以pET30a为载体构建pET30a-AmPR-10;③以pET30a为载体、大肠杆菌分子伴侣skp修饰构建pET30a-skp-AmPR-10。通过SDS-PAGE分析,目的蛋白可溶性表达量比较结果是：pET30a-skp-AmPR-10 > pET30a-AmPR-10 > pET28a-AmPR-10。由蛋白质三级结构模拟分析发现,载体pET-30a上的标签S-tag通过影响AmPR-10局部α螺旋构象而提高目标蛋白的可溶性表达,分子伴侣skp通过提高融合蛋白整体α螺旋比例进一步提升目标蛋白可溶性表达量。研究解决了目前从天然黄芪中提取蛋白操作复杂、提取量小的问题。同时,经测定,目的蛋白具有核酸酶活性,为外源AmPR-10相关活性研究提供了依据。     

Patterns of Expression in the Matrix Proteins Responsible for Nucleation and Growth of Aragonite Crystals in Flat Pearls of Pinctada fucata

The initial growth of the nacreous layer is crucial for comprehending the formation of nacreous aragonite. A flat pearl method in the presence of the inner-shell film was conducted to evaluate the role of matrix proteins in the initial stages of nacre biomineralization in vivo. We examined the crystals deposited on a substrate and the expression patterns of the matrix proteins in the mantle facing the substrate. In this study, the aragonite crystals nucleated on the surface at 5 days in the inner-shell film system. In the film-free system, the calcite crystals nucleated at 5 days, a new organic film covered the calcite, and the aragonite nucleated at 10 days. This meant that the nacre lamellae appeared in the inner-shell film system 5 days earlier than that in the film-free system, timing that was consistent with the maximum level of matrix proteins during the first 20 days. In addition, matrix proteins (Nacrein, MSI60, N19, N16 and Pif80) had similar expression patterns in controlling the sequential morphologies of the nacre growth in the inner-film system, while these proteins in the film-free system also had similar patterns of expression. These results suggest that matrix proteins regulate aragonite nucleation and growth with the inner-shell film in vivo.     

LjMOT1, a high‐affinity molybdate transporter from Lotus japonicus,is essential for molybdate uptake,but not for the delivery to nodules

Molybdenum (Mo) is an essential nutrient for plants, and is required for nitrogenase activity of legumes. However, the pathways of Mo uptake from soils and then delivery to the nodules have not been characterized in legumes. In this study, we characterized a high‐affinity Mo transporter (LjMOT1) from Lotus japonicus. Mo concentrations in an ethyl methanesulfonate–mutagenized line (ljmot1) decreased by 70–95% compared with wild‐type (WT). By comparing the DNA sequences of four AtMOT1 homologs between mutant and WT lines, one point mutation was found in LjMOT1, which altered Trp²⁹² to a stop codon; no mutation was found in the other homologous genes. The phenotype of Mo concentrations in F₂ progeny from ljmot1 and WT crosses were associated with genotypes of LjMOT1. Introduction of endogenous LjMOT1 to ljmot1 restored Mo accumulation to approximately 60–70% of the WT. Yeast expressing LjMOT1 exhibited high Mo uptake activity, and the K_m was 182 nm . LjMOT1 was expressed mainly in roots, and its expression was not affected by Mo supply or rhizobium inoculation. Although Mo accumulation in the nodules of ljmot1 was significantly lower than that of WT, it was still high enough for normal nodulation and nitrogenase activity, even for cotyledons‐removed ljmot1 plants grown under low Mo conditions, in this case the plant growth was significantly inhibited by Mo deficiency. Our results suggest that LjMOT1 is an essential Mo transporter in L. japonicus for Mo uptake from the soil and growth, but is not for Mo delivery to the nodules.     

曾候乙墓穴木椁微生物的分离与鉴定

从曾侯乙墓中椁室、东椁室、西椁室和北椁室分别取样,经平板划线培养,分离,纯化共获得典型的菌株16株。将16株分别涂片镜检,生理生化分析鉴定,结果证明芽孢杆菌属11株,其中苏云金变种1株,地衣芽孢杆菌1株,巨大芽孢杆菌1株,球形芽孢杆菌5株,蜡状芽孢杆菌2株,多粘芽孢杆菌1株。其余菌株微杆菌属4株,黄色杆菌属黄杆菌1株。     

曾候乙墓穴木椁微生物降解对木材危害及防治措施研究

从曾候乙墓分离到的16株菌,其中有11株为芽孢杆菌属类,4株微杆菌属,1林为黄色杆菌属。将16株细菌分别侵染6种木材,结果证明细菌降解对6种木材均有不同的韧性下降,个别木材出现腐烂病灶。2号菌降解后对泡桐、枣树、马尾松、香樟、刺槐和桑树分别下降8％,12％,10％,5％,10％和13％。3,5,6,7,8,9和16号菌对上述6种木材也有不同程度影响。而4,11,12,13,14和15号菌对6种木材的韧性下降影响不明显。但其渗透性大大提高了,因此也会影响木材的使用年限。采用杀菌、杀虫的石油气,乙醇等有机溶济,防治微生物降解产物对木材的浸蚀,均有较好的防菌、防认腐效果。