氧化还原对成年大鼠海马CA1区锥体神经元外向整流氯通道的调控作用

采用膜片钳内面向外式记录技术,研究急性分离成年大鼠海马CAl区锥体神经元外向整流氯离子通道的氧化还原调控。发现细胞内侧给予氧化剂DTNB(5,5'-dithiobis-2-nitrobenzoic acid),可显著减弱氯通道的活动,IC50值为(28.05±2.42)μmol/L;还原剂DTT(dithiothreitol)对氯通道没有明显影响,但可逆转DTNB引起的抑制效应。说明DTNB不改变通道电导,其引起的通道活动减弱是由氯通道关闭时间延长而开放时间缩短所致。研究还发现,另一对氧化型和还原型谷胱甘肽具有与DTNB和DTT同样的效应。本研究结果显示,成年大鼠海马CA1区锥体神经元外向整流氯通道可以被细胞内氧化还原剂所调控。     

Activation of STAT3 stimulates AHSP expression in K562 cells

Studies on the chaperone proteinα-hemoglobin stabilizing protein(AHSP)reveal that abundant AHSP in erythroid cells enhance the cells’tolerance to oxidative stress imposed by excessα-hemoglobin in pathological conditions.However,the potential intracellular modulation of AHSP expression itself in response to oxidative stress is still unknown.The present study examined the effect and molecular mechanism of STAT3,an oxidative regulator,on the expression of AHSP.AHSP expression increased in K562 cells upon cytokine IL-6-induced STAT3 activation and decreased in STAT3 knock-down K562 cells.Regulation of AHSP in oxidative circumstance was then examined inα-globin-overloaded K562 cells,and real-time PCR showed strengthened expression of both AHSP and STAT3.ChIP analysis showed binding of STAT3 to AHSP promoter and binding was significantly augmented with IL6 stimulation and uponα-globin overexpression.Dual luciferase reporter assays of the wildtype and mutated SB3 element,an IL-6RE site,in the AHSP promoter in K562 cells highlighted the direct regulatory effect of STAT3 on AHSP gene.Finally,direct binding of STAT3 to SB3 site of AHSP promoter was confirmed with EMSA assays.Our work reveals an adaptive AHSP regulation mediated by the redox-sensitive STAT3 signaling pathway,and provides clues to the therapeutic strategy for AHSP enhancement.     

植物体细胞胚发生的分子基础①

ＭｏｌｅｃｕｌａｒＦｏｕｎｄａｔｉｏｎｉｎＰｌａｎｔＳｏｍａｔｉｃＥｍｂｒｙｏｇｅｎｅｓｉｓＸＩＮＧＧｅｎｇ－ＳｈｅｎｇＣＵＩＫａｉ－ＲｏｎｇＳＨＡＮＬｕｎＷＡＮＧＹａ－Ｆｕ（ＴｈｅＳｔａｔｅＫｅｙＬａｂｏｒａｔｏｒｙｏｆＡｒｉｄＡｇｒｏｅｃｏｌｏｇ...     

线粒体DNA缺失与神经肌肉性疾病的研究

应用PCR技术对13例神经肌肉疾病患者的线粒体DNA缺失进行了研究。结果表明,其中二例肢带型肌营养不良症患者骨骼肌组织和一例帕金森氏病患者的血细胞线粒体DNA中存在至少526bp的缺失。提示线粒体突变在一些神经肌肉性疾病的发生中起一定的作用。 Abstract: Using PCR technique,we analysed the skeletal muscle and blood of 13 patients with neuromuscular disaeases.The results show that a mutant mitochondrial DNA with at least 526bp deletion exits in the skeletal muscles of 2 patients with Erb muscular dystrophy and in the blood of a patients with Parkinson’s disease.From our results,mitochondrial DNA mutations could be an important contributory factor to neuromuscular diseases.     

离子注入对微生物细胞的刻蚀与对DNA的损伤及修复

以耐辐射异常球菌为试材,以E. coli 为对照,用显微扫描电镜和3H-TdR标记,研究了离子注入对微生物细胞的刻蚀与对DNA的损伤及其修复。结果表明,注入离子对细胞存在着刻蚀损伤;中性蔗糖梯度密度离心沉降分析证明, 大剂量下离子注入可直接导致DNA损伤,并观察到在对应的存活率峰值注入剂量下,D. radiodurans修复损伤DNA的能力比E. coli 强,还证明了细胞经不同时间温育后,损伤的DNA分子得到了部分修复。 Abstract：　The direct action of N+implantationin on D. radioduransand E. coliwas investigated by SEM, and their cells were labeled with 3H-TdR, which were implanted by 20keV N+after incubation 18hours, then the DNA of lysed cells was subjected to the neutral sucrose gradient(5%～20%) ultra-centrifugation sedimentation analysis. The results showed that N+implantation exerted direct action on two kinds of microorganisms; the momentum transfer and energy deposition of implantation ions produced the direct etching damage on cells, and repair DNA efficiency of D.radiodurans was higher than that of E. coli. Meanwhile, the damaged DNA incomplete repairing was observed. When incubation was continued up to 6 hours, the rejoined DNA molecules broke again. The repair of damaged DNA could be inhibited by 200μg/ml chloramphenicol. This suggested that DNA damage was serious by ion implantation and damaged DNA repair of cells need continuously synthesizing repair enzyme.     

In vitro Anti-viral Activity of the Total Alkaloids from Tripterygium hypoglaucum against Herpes Simplex Virus Type 1

Herpes simplex virus type 1 (HSV-1) is a commonly occurring human pathogen worldwide. There is an urgent need to discover and develop new alternative agents for the management of HSV-1 infection. Tripterygium hypoglaucum (level) Hutch (Celastraceae) is a traditional Chinese medicine plant with many pharmacological activities such as anti-inflammation, anti-tumor and antifertility. The usual medicinal part is the roots which contain about a 1% yield of alkaloids. A crude total alkaloids extract was prepared ...     

A Stable Upstream Stem-loop Structure Enhances Selection of the First 5'-ORF-AUG as a Main Start Codon for Translation Initiation of Human ACAT1 mRNA

Acyl-coenzyme A:cholesterol acyltransferase (ACAT)is an integral membrane protein, which is mainly locatedin rough endoplasmic reticulum (ER), and is responsiblefor catalyzing the intracellular formation of cholesterylester from cholesterol and long-chain fatty acyl-coenzymeA [1,2]. Human ACAT1 cDNA K1 was firstly cloned andfunctionally expressed in 1993 [3]. Further studies withspecific anti-ACAT1 antibody (DM10) illustrated that onemajor 50 kD ACAT1 protein was expressed in various…     

浙北地区常见绿化树种光合固碳特征

高固碳能力的树种选择是营造优质碳汇林,发展碳汇林业的重要基础工作.以浙北地区常见的30种造林绿化树种为研究材料,利用LI-6400便携式光合测定仪,测定树木光合日变化及不同光强梯度下光合作用的光响应特性,并根据实验观测值进行计算,对30个树种的日净固碳量和光合生理拟合参数进行Ward法聚类分析和因子分析.结果表明:香樟的固碳量最大((11.374±1.020) g·m-2·d-1),其次为碧桃、垂柳、石栎、无患子,固碳量最小的为红叶李((2.178±0.605) g·m-2·d-1),香樟和红叶李的日净固碳量有极显著差异(P＜0.01);树木的生理特性指标分析进一步反映了树种在浙北地区生长适应性及固碳能力大小,同时,根据树木的生理特性指标进行因子分析和聚类分析的结果,香樟、碧桃在浙北地区生长适应性较好,其次为无患子、垂柳、女贞等;根据树种固碳量及生理指标综合测定分析,建议在浙北地区造林绿化中可以优先选用香樟、碧桃、垂柳、无患子、石栎、女贞这些树种.     

过氧化氢驱动的细胞色素P450酶研究进展

细胞色素P450酶在自然界中广泛存在,能催化多种类型的氧化反应,在有机合成和生物化工方面具有重要的应用潜力。尽管大多数P450酶通常需要辅酶和复杂的电子传递体系协助活化氧分子,一些P450酶也可以利用过氧化氢作为末端氧化剂,这极大地简化了催化循环,为P450酶的合成应用提供了一条新的简便途径。本文系统地介绍了几类过氧化氢驱动的P450酶催化体系,包括脂肪酸羟化酶P450SPα和P450BSβ、脂肪酸脱羧酶P450OleTJE、人工改造的羟化酶P450BM3和P450cam突变体、以及基于底物误识别策略的P450-H2O2体系。通过分析催化反应机制,本文探讨了P450-H2O2催化体系在目前存在的挑战和可能的解决途径,并对其进一步应用前景进行了展望。     

大鼠全脑缺血后海马CA1区锥体神经元L型钙通道电流的改变(英)

已有研究表明在脑缺血期间及再灌流后早期,海马CA1锥体神经元细胞内钙浓度明显升高,这一钙超载被认为是缺血性脑损伤的重要机制之一.电压依赖性钙通道是介导正常CA1神经元钙内流的主要途径.实验观察了脑缺血再灌流后早期海马CA1锥体神经元电压依赖性L型钙通道的变化.以改良的四血管闭塞法制作大鼠15 min前脑缺血模型,在急性分离的海马CA1神经元上,采用膜片钳细胞贴附式记录L型电压依赖性钙通道电流.脑缺血后CA1神经元L型钙通道的总体平均电流明显增大,这是由于通道的开放概率增加所致.进一步分析单通道动力学显示,脑缺血后通道的开放时间变长,通道的开放频率增大.研究结果提示L型钙通道功能活动增强可能参与了缺血后海马CA1锥体神经元的细胞内钙浓度升高.