A novel technique to specifically analyze the secretome of cells and tissues

The secretome of cells and tissues may reflect a broad variety of pathological conditions and thus represents a rich source of biomarkers. The identity of secreted proteins, usually isolated from cell supernatants or body fluids, is hardly accessible by direct proteome analysis, because these proteins are often masked by high amounts of proteins actually not secreted by the investigated cells. Here, we present a novel method for the specific detection of proteins secreted by human tissue specimen as well as cultured cells and chose liver as a model. The method is based on the metabolic labelling of proteins synthesized during a limited incubation period. Then, the cell supernatant is filtered, precipitated, and subjected to two-dimensional gel electrophoresis. Whereas fluorography detected a large number of proteins derived from residual plasma and dead cells, the autoradiographs selectively displayed genuinely secreted proteins. We demonstrate the feasibility of this approach by means of the secretomes of the hepatocellular carcinoma-derived cell line HepG2 and human liver slices. The selective identification of cell- and tissue-specific protein secretion profiles may help to identify novel sets of biomarkers for wide clinical applications.     

基于DEMATEL和ANP的电子商务代运营服务企业核心竞争力影响因素研究

随着“互联网+”与传统行业的不断融合,越来越多自身电子商务运营能力不足的中小企业选择电子商务服务外包的方式进入电子商务行业,电子商务代运营服务企业面临机遇的同时也面临激烈的竞争与挑战。为提高电子商务代运营服务企业的竞争力,提升其核心能力建设,从企业综合实力、代运营技术、代运营服务项目、代运营服务质量、人力资源、企业信誉、发展潜力7个方面构建了电子商务代运营服务商竞争能力评价指标体系。针对评价指标众多,主观、客观因素繁杂的状况,建立了DEMATEL与ANP相结合的多属性综合决策方法,首先,使用DEMATEL分析评价指标间的影响关系,绘制影响因素四分图;其次,构建了电子商务代运营服务商竞争能力重要性及关联性ANP网络,引入ANP方法计算各指标的全局权重,并与调整后的综合矩阵混合计算;最后,通过分析混合权重,识别出管理体系、财务状况、历史业绩、服务理念、战略规划为电子商务代运营企业核心竞争力影响的关键因素,进而制定培育企业核心竞争能力的对策。     

空间机器人组装超大型结构的动力学分析北大核心CSCD

超大型航天结构具有超大柔性、超低固有频率的特点,空间机器人在轨组装时应尽可能避免激起超大型结构的柔性振动.空间机器人组装超大型结构模块的过程分成抓捕阶段、位姿调整与稳定阶段、安装阶段和爬行阶段.通过对安装阶段的动力学与控制研究,提出共线安装的轨迹规划方法,有效避免了柔性结构振动.首先,采用自然坐标法和绝对节点坐标法建立主结构-空间机器人-待组装结构的在轨组装系统动力学模型.然后,将共线安装的要求转化为空间机器人的轨迹规划约束,要求空间机器人质心到主结构/待组装结构的距离保持不变,实现共线安装的轨迹规划.数值仿真表明:提出的组装方法在组装过程中可有效避免超大型结构的横向运动,降低夹持力矩.最后,分析了系统参数对组装过程动力学响应的影响,为超大型航天器的在轨组装提供了参考.     

“限时服务规则”下的复线船闸多目标调度优化

针对现有船舶过闸排队规则的欠缺,基于“限时服务规则”,构建复线船闸多目标双层优化调度模型:上层模型用于获得两个闸室安全区域的船舶排布可行方案;下层模型用于获得不同船舶排布可行方案的优化闸次数。下层模型分两个阶段完成:对符合“限时服务规则”的船舶,构建以闸次最少为目标的0-1规划模型,获得此类船舶安排的闸次;对其余船舶按照“先到先服务规则”,构建以闸次最少、闸室利用率最大为目标的多目标决策模型,获得不同船舶排布可行方案应该安排的频次。以位于江苏省干线航道上的某复线船闸某日24小时内过闸船舶的数据为例,计算结果表明:采用本文优化模型获得的优化方案与“经验编排方式”相比,两座船闸各节约2个闸次,两个船闸的平均闸室利用率分别提高了3.66和4.72个百分点。     

市场化改革背景下养老机构服务供给博弈研究

考虑政府补贴、市场化程度、养老机构服务成本、老年人偏好等因素,构建了完全国有化、公私并存的半市场化和完全市场化三种情形下公办和民办养老机构之间的两阶段博弈模型,比较分析了市场化程度对养老机构服务价格制定和服务水平选择的影响,实证讨论了各参数变化对养老机构收益及社会福利的影响。研究表明:完全市场化机制会降低养老机构服务质量,完全国有化机制会陷入社会福利陷阱,探索公私并存的最佳市场化水平才能更好地提高社会福利;适度推进市场化改革,提高养老机构服务水平,降低民办养老机构服务成本,适度提高对民办养老机构的补贴,将有效缓解公办和民办养老机构的不公平竞争;提高老年人支付能力,推进养老机构供给侧结构性改革,将保障机构养老服务的有效供给,促进养老服务业的健康有序发展。     

一种新的配电网故障恢复算法及其改进

当配电网中某运行设备发生故障并切除后,会引起一些负荷失去供电,此时需要快速有效地寻找恢复供电路径。文中介绍的算法根据配电网的辐射状结构特征,把电源点和失去供电的负荷点看作“兴趣点”,把寻找供电路径转化为求解给定图的“兴趣树”,从而简化了问题。计算结果表明,经多方面的改进后,算法更加快速有效,在配电系统自动化高层软件开发中,有在线应用前景,本算法适用于负荷集中的１０ｋＶ以上配电网。     

科学发现及其逻辑性研究

在讨论科学发现,科学发现逻辑性研究的历史的基础上,就科学发现逻辑性研究的两个新视角作了评价和展望。“信息域”拓展了科学发现逻辑性的空间,突现出社会文化因素的意义;模糊思维把科学发现的逻辑性研究转换为对模糊信息进行加工、度量的认知过程,强化了科学发现逻辑性研究的现实性。     

Site-specific detection of S-nitrosylated PKB alpha/Akt1 from rat soleus muscle using CapLC-Q-TOF(micro) mass spectrometry

Protein Kinase Balpha(PKBalpha, or Akt1) is believed to play a crucial role in programmed cell death, cancer progression and the insulin-signaling cascade. The protein is activated by phosphorylation at multiple sites and subsequently phosphorylates and activates eNOS. Free cysteine residues of the protein may capture reactive, endogenously produced nitric oxide (NO) as S-nitrosothiols. Site-specific detection of S-nitrosylated cysteine residues, usually at low stoichiometry, has been a major challenge in proteomic research largely due to the lack of mass marker for S-nitrosothiols that are very labile under physiologic conditions. In this report we describe a sensitive and specific MS method for detection of S-nitrosothiols in PKB alpha/Akt1 in rat soleus muscle. PKB alpha/Akt1 was isolated by immunoprecipitation and 2D-gel electrophoresis, subjected to in-gel tryptic digestion, and cysteinyl nitrosothiols were reacted with iodoacetic acids [2-C(12)/C(13) = 50/50] under ascorbate reduction conditions. This resulted in the production of relatively stable carboxymethylcysteine (CMC) immonium ions (m/z 134.019 and m/z 135.019) within a narrow argon collision energy (CE = 30 +/- 5 V) in the high MS noise region. In addition, free and disulfide-linked cysteine residues were converted to carboxyamidomethylcysteines (CAM). Tryptic S-nitrosylated parent ion was detected with a mass accuracy of 50 mDa for the two CMC immonium ions at the triggered elution time during capillary liquid chromatography (LC) separation. A peptide containing Cys(296) was discriminated from four co-eluting tryptic peptides under lock mass conditions (m/z 785.8426). S-nitrosothiol in the tryptic peptide, ITDFGLBKEGIK (B: CAM, [M + 2H](2+) = 690.86, Found: 690.83), is believed to be present at a very low level, since the threshold for the CMC immonium trigger ions was set at 3 counts/s in the MS survey. The high levels of NO that are produced under stress conditions may result in increased S-nitrosylation of Cys(296) which blocks disulfide bond formation between Cys(296) and Cys(310) and suppresses the biological effects of PKB alpha/Akt1. With the procedures developed here, this process can be studied under physiological and pathological conditions.     

化合物库的组合技巧和组合合成

从化合物库的组合技巧和组合合成方法两个方面探讨了组合化学及其在药物开发中的应用。     

RNA Methylation m6A: A New Code and Drug Target?

The m⁶A‐RNA modification is a dynamic and reversible process, which has emerged as a new RNA code for the regulation of gene expression. The functional network of methyltransferases (writers), demethylases (erasers), and binding proteins (readers) modulate the level of m⁶A modification. Dysfunction of RNA methylation has been associated with various fundamental biological processes and human diseases. Herein, we briefly introduce an understanding‐enabled manipulation on m⁶A‐RNA modification with an emphasis on the use of small‐molecule intervention.