Eukaryotic expression and biological activity analysis of neuroprotective peptide [Gly14]-Humanin

Objective To investigate the expression of neuroprotective peptide [Gly14]-Humanin (HNG) in eukaryotic cells by gene engineering technique and analyze its biological activity. Methods By means of asymmetrical primer/template, double stranded cDNA of HNG with FLAG in its C-terminal was obtained, which was cloned into the plasmid pcDNA3.1(-), and the resultant recombinant vector pcDNA3.1(-)/HNG-FLAG was transfected into PC12 cells. At the same time, the recombinant vector pcDNA3.1(-)/EGFP was transfected to control the efficiency of transfection. The expression of HNG in the cells was determined by immunocytochemistry. In order to analyze the biological activity of the expressed HNG, 25μM Aβ25-35 peptide was added to the culture medium of the transfected cells for 24h, then cell morphology, MTT assay and Hoechst 33258 staining were observed. Results The eukaryotic expression vector of pcDNA3.1(-)/HNG-FLAG was identified by enzyme digestion and sequencing. HNG was highly expressed in PC12 cells. After exposure of PC12 cells to 25μM Aβ25-35 for 24h, cell viability decreased to (65.8±5.3)%, and the dystrophic changes of neuritis and nuclei condensation were obvious. When cells were pre-transfected with pcDNA3.1(-)/HNG-FLAG, Aβ25-35-induced cell death and morphological changes of cells and nuclei were suppressed. In contrast, pre-transfected with empty vector did not protect cells from Aβ25-35-induced toxicity. Conclusion The eukaryotic expression vector for FLAG-tagged HNG was successfully constructed and expressed in PC12 cells. Expressed HNG has biological activity.     

Analysis of within-household effects and between-household differences in maintenance activity allocation

This paper investigates the allocation of household individuals to out-of-home maintenance activities using the rich activity-travel diary data from the San Francisco Bay Area. Two inter-related decisions are considered in this context: (i) whether the given activity episode is performed individually (solo) or jointly, and (ii) the person who participates in the activity, if it is a solo activity. To account for the conditional nature of the solo activity person selection, a nested mixed logit modeling framework is proposed and implemented to jointly analyze person allocation for all maintenance activities performed by a household on a given day. The model is used to investigate within-household effects and between-household differences. The proposed model relaxes some important restrictions in person allocation models by accounting for various sources of correlations and relaxing the assumption of constant variance across households. The proposed model is used to analyze the differences in person allocation between different types of households. The results indicate that life-cycle and household role, income, gender, employment status, and several types of constraints (activities including cost, time-availability, vehicle-availability, coordination constraints, and child-care obligations) affect person allocation decisions in the context of maintenance activities. The empirical results indicate the presence of various sources of correlations across persons, over activities, and within-household that are significant. In addition, the data also provides evidence that the unobserved variances in person selection utilities are not constant across households. A better understanding of these within-household interactions and between-household differences may be used in activity-based simulation models and to develop more effective and focused demand management measures.     

客车品牌整合营销传播中的项目管理应用研究

将项目管理的思想引入到客车行业的品牌整合营销传播之中,重点分析研究客车品牌整合传播中的进度控制和成本管理,以提高客车品牌营销的管理水平和传播效果。     

山区河流泥石流灾害防治技术研究

为改善通航条件、提高航道等级,对山区河流泥石流灾害防治技术进行了研究.根据形成机理、运动规律、成灾机制、岸坡的地形地貌特点,把山区河流型泥石流分为坡面流和沟谷流;基于土质结构原理与岩土稳定理论,提出了修复植被、规范人类行为与截流排水、拦粗排细、转向导流等工程防治技术相结合的山区河流泥石流灾害的综合防治技术.结果表明,该...     

电沉积Ni-S、Ni-Co-S合金析氢阴极的研究

在电沉积法制备Ni-S合金电极的基础上,向镀液中添加硫酸钴制备了Ni-Co-S合金电极。电化学测试结果表明,Ni-Co-S合金电极和Ni-S合金电极的催化性能都要好于其它电极,Ni-Co-S合金电极的性能随着硫酸钴浓度的升高先升高后降低。计算了电极的电极反应动力学参数(包括Tafel斜率b、交换电流密度i0和过电位),解释了Ni-Co-S合金电极和Ni-S合金电极在不同电流密度区活性不同的原因。恒电流电解表明,Ni-Co-S合金电极具有较高的稳定性。电极的微观形貌用SEM进行表征,成份用能谱进行分析,X射线衍射分析电极的晶型结构。     

Estimating a model of dynamic activity generation based on one-day observations: Method and results

In this paper we develop and explore an approach to estimate dynamic models of activity generation on one-day travel-diary data. Dynamic models predict multi-day activity patterns of individuals taking into account dynamic needs as well as day-varying preferences and time-budgets. We formulate an ordered-logit model of dynamic activity-agenda-formation decisions and show how one-day observation probabilities can be derived from the model as a function of the model’s parameters and, with that, how parameters can be estimated using standard loglikelihood estimation. A scale parameter cannot be identified because information on within-person variability is lacking in one-day data. An application of the method to data from a national travel survey illustrates the method. A test on simulated data indicates that, given a pre-set scale, the parameters can be identified and that estimates are robust for a source of heterogeneity not captured in the model. This result indicates that dynamic activity-based models of the kind considered here can be estimated from data that are less costly to collect and that support the large sample sizes typically required for travel-demand modeling. We conclude therefore that the proposed approach opens up a way to develop large-scale dynamic activity-based models of travel demand.     

Rural transport – Valuing the mobility of older people

This paper describes a series of focus groups that were undertaken in the Republic of Ireland and Northern Ireland to examine the challenges faced by older people in rural Ireland when trying to make trips. The paper outlines transport policy in both Northern Ireland and the Republic of Ireland and describes the activity patterns of older trip makers and the modes they use. It also discusses some of the contrasts between challenges faced by older men and older women. Both older men and older women experience a shrinking in activity spaces and activity participation when older. Older women are less likely to drive and to own a car and so are seriously impacted by loss of a spouse in terms of their unmet travel needs. However, older men also experience difficulties: the alternatives that are available (such as community transport schemes) provide trips and services that are more attractive to women. In addition, the car dependent lifestyles of men throughout their adult lives seems to make them less prepared for life without a car than older women.     

铁路现代物流营销模式的发展目标和实现机制研究——以铁路钢铁供应链为例

铁路现代物流必须实现从过去单个运输职能向组成供应链的多元素职能转换。集成化策略是供应链发展的最高层次,通过成员企业之间的协同合作,对供应链上、下游企业的整体集成可以为客户创造低成本高效能的最大化价值。多快好省应当是包括铁路钢铁供应链在内的所有供应链集成化目标,也是以供应链理论为基础的铁路现代物流营销模式的发展目标。     

“互联网 +”背景下汽车营销管理模式的创新

信息技术的不断普及大大地拓宽了大众获取咨询的方式和渠道,人们的选择也随之变得多种多样,因此人们倾向于从网络渠道获取相关的信息,比如说汽车的价格、质量以及品牌。同时,汽车企业也需要及时迎合这股互联网的热潮,利用互联网的高效性、便捷性,创新相关汽车营销和发展模式,让网络成为顾客和商家之间的沟通桥梁,以此推广企业的产品,扩大汽车销售市场,提高产业的经济效益和企业竞争力,从而占据一定的汽车销售市场。基于此,本文特地研究了“互联网+”背景下的汽车营销管理模式,意图给相关的营销管理提供一定参考。     

社会主义国家对外开放理论的飞跃

对外开放理论是邓小平理论的重要组成部分。它不仅在实践上指导中国在党的十一届三中全会以后取得了对外经济联系及整个社会经济的重大发展,同时在总结前人经验教训的基础上,结合中国及世界的最新发展,实现了对社会主义国家开放理论的飞跃和升华。