白血病K562细胞株中CD34~+细胞群的分离及其生物学特性

目的分离白血病K562细胞株中CD34+细胞群,并分析其生物学特征,为从干细胞角度治疗白血病提供实验依据。方法采用免疫磁性分选法分离K562细胞中CD34+细胞群,台盼蓝拒染法检测细胞活性;流式细胞术检测CD34+细胞比例及细胞周期;单细胞克隆培养检测CD34+细胞自我更新能力;RT-PCR法检测CD34+细胞分化相关指标促红细胞生成素(Erythropoietin,EPO)和粒细胞-巨噬细胞集落刺激因子(Granulocyte macrophage colony stimulatingfactor,GM-CSF)基因mRNA的转录水平;并检测CD34+细胞耐药蛋白P-gp(P-glycoprotein)的表达。结果免疫磁性分选法可有效分离出CD34+细胞群,细胞活性为99%～100%;分离的CD34+细胞含量占细胞总数的78.5%～85.3%,细胞大部分处于静止状态,G0/G1期细胞比例达80%左右,显著高于分离前的K562细胞;CD34+细胞群具有形成混合集落的能力;与K562细胞相比,CD34+细胞EPO和GM-CSF基因mRNA的转录水平明显下降(P<0.01),P-gp表达阳性。结论成功从K562细胞株中分离了CD34+细胞群,其具有自我更新和多向分化的能力,表明其具有白血病干/祖细胞的生物学特点。     

100G OTN大容量传输技术解析

从OSNR性能改善、色散(CD)容限、偏振模色散(PMD)容限、相位调制、偏振复用等方面介绍了100G PM-QPSK关键技术特性,最后给出了烽火通信推出的100G大容量传输系统的解决方案.     

Misfolding and Amyloid Aggregation of Apomyoglobin

Apomyoglobin is an excellent example of a monomeric all α-helical globular protein whose folding pathway has been extensively studied and well characterized. Structural perturbation induced by denaturants or high temperature as well as amino acid substitution have been described to induce misfolding and, in some cases, aggregation. In this article, we review the molecular mechanism of the aggregation process through which a misfolded form of a mutated apomyoglobin aggregates at physiological pH and room temperature forming an amyloid fibril. The results are compared with data showing that either amyloid or aggregate formation occurs under particular denaturing conditions or upon cleavage of the residues corresponding to the C-terminal helix of apomyoglobin. The results are discussed in terms of the sequence regions that are more important than others in determining the amyloid aggregation process.     

Serum Oxidant and Antioxidant Status Following an All-Out 21-km Run in Adolescent Runners Undergoing Professional Training—A One-Year Prospective Trial

This study investigated the 1-year longitudinal effect of professional training in adolescent runners on redox balance during intense endurance exercise. Changes in selected serum oxidant and antioxidant status in response to a 21-km running time trial in 10 runners (15.5 ± 1.3 years) undergoing professional training were evaluated twice in 12 months (pre- and post-evaluation). Venous blood samples were collected immediately before and 4-h following the 21-km run for analysis of serum concentrations of thiobarbituric acid-reactive substances (TBARS), xanthine oxidase (XO), catalase (CAT), reduced glutathione (GSH), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC). In pre-evaluation trial, serum TBARS and SOD decreased after the 21-km run (p < 0.05) while XO, GSH, CAT and TAOC were unchanged. In post-evaluation trial, serum TBARS and SOD decreased, whereas XO and CAT increased post-exercise (p < 0.05). Furthermore, pre-exercise serum T-AOC, post-exercise serum XO, CAT, T-AOC (p < 0.05), and GSH (p = 0.057) appeared to be higher than the corresponding pre-evaluation values. The current findings suggest that a professional training regime in adolescent runners is not likely to jeopardize the development of their antioxidant defense. However, uncertainties in the maintenance of redox balance in runners facing increased exercise-induced oxidative stress as a consequence of training-induced enhancement of exercise capacity await further elucidation.     

Green Fluorescent Protein (GFP)-Based Overexpression Screening and Characterization of AgrC,a Receptor Protein of Quorum Sensing in Staphylococcus aureus

Staphylococcus aureus AgrC is an important component of the agr quorum-sensing system. AgrC is a membrane-embedded histidine kinase that is thought to act as a sensor for the recognition of environmental signals and the transduction of signals into the cytoplasm. However, the difficulty of expressing and purifying functional membrane proteins has drastically hindered in-depth understanding of the molecular structures and physiological functions of these proteins. Here, we describe the high-yield expression and purification of AgrC, and analyze its kinase activity. A C-terminal green fluorescent protein (GFP) fusion to AgrC served as a reporter for monitoring protein expression levels in real time. Protein expression levels were analyzed by the microscopic assessment of the whole-cell fluorescence. The expressed AgrC-GFP protein with a C-terminal His-tagged was purified using immobilized metal affinity chromatography (IMAC) and size exclusion chromatography (SEC) at yields of ≥10 mg/L, following optimization. We also assessed the effects of different detergents on membrane solubilization and AgrC kinase activity, and polyoxyethylene-(23)-lauryl-ether (Brij-35) was identified as the most suitable detergent. Furthermore, the secondary structural stability of purified AgrC was analyzed using circular dichroism (CD) spectroscopy. This study may serve as a general guide for improving the yields of other membrane protein preparations and selecting the appropriate detergent to stabilize membrane proteins for biophysical and biochemical analyses.     

一种基于谱分析与改进方位非线性变标的大斜视聚束成像算法

针对大斜视聚束SAR成像中方位频谱混叠,且距离向与方位向严重耦合的问题,该文提出一种利用谱分析(SPECtral ANalysis, SPECAN)解方位频谱混叠、距离向采用调频变标(Chirp Scaling, CS)、方位向采用改进非线性变标(Extended Non-linear Chirp Scaling, ENCS)的大斜视聚束成像算法,首先将回波信号走动校正,然后利用SPECAN消除方位频谱混叠。进行距离徙动校正(Range Cell Migration Correction, RCMC)之后,利用CS消除距离弯曲的空变性,方位向采用 ENCS 补偿沿方位向变化的多普勒调频率,从而有效地提高了方位向的聚焦深度。仿真结果和分析表明,该方法能够在较大斜视角的聚束SAR模式下得到聚焦良好的高分辨率场景。     

深圳大运中心主体育场单层折面空间网格结构临时支撑卸载技术

针对深圳大运中心主体育场钢结构工程单层折面空间网格的结构特征和临时支撑体系布局,分析了不同卸载工况对结构产生的影响,综合等比例卸载和等距离卸载方法的优点,采取大吨位千斤顶和计算机同步控制卸载技术,成功解决多点高行程卸载和80个千斤顶同步控制的难题。     

木结构火灾性能研究进展

在已有木结构火灾性能研究资料基础上,分析和总结了木结构火灾特征、木材炭化机理和微观结构变化、木材炭化速度模型、受火木构件剩余承载力、持荷木构件耐火极限等研究现状,并提出了需要进一步研究的重点。