SNAP-23蛋白在鲈鱼巨噬细胞白细胞介素1β分泌过程中的功能

SNARE蛋白家族是所有真核细胞胞吐及分泌作用中的关键因子，由其介导的运输囊泡膜与靶膜的锚靠、融合为胞内蛋白的运出提供了一条重要途径。体外试验表明，Syntaxin6-Syntaxin7-Vti1b，SNAP-23-Syntaxin4等SNARE核心蛋白之间精确的相互作用是哺乳动物巨噬细胞肿瘤坏死因子α (TNF-α)运输和分泌的必备条件，在机体非特异性免疫应答反应过程中起重要作用。 本研究受上述启示，旨在揭示SNARE蛋白在海洋鱼类免疫细胞内重要细胞因子白细胞介素1β (IL-1β)的分泌过程中的作用。参照Percoll密度梯度离心技术，从鲈鱼头肾组织分离纯化巨噬细胞进行稳定培养；利用RT-PCR方法克隆出鲈鱼t-SNARE蛋白SNAP-23和Syntaxin3的部分cDNA序列，再结合先前克隆的VAMP2和已知的鲈鱼IL-1β，TNF-α和IL-8的基因序列，设计特异性引物。利用Real-time PCR技术在mRNA水平上精确测定鲈鱼巨噬细胞中上述6种基因在革兰氏阴性菌脂多糖(LPS)分子刺激下的表达变化，发现SNAP-23基因与三种细胞因子基因的表达正相关；通过免疫印迹检测SNAP-23蛋白表达变化，利用酶联免疫吸附试验(ELISA)检测IL-1β的分泌水平，在蛋白水平上验证了SNAP-23表达与IL-1β分泌的正相关性；利用5`RACE和3`RACE技术克隆出鲈鱼SNAP-23全长基因，结合定点突变策略和靶向PCR克隆手段，构建鲈鱼SNAP-23野生型融合质粒pEGFP-SNAP-23wt，Cys缺失突变融合质粒pEGFP-SNAP-23ΔCys和模拟E型肉毒神经毒素(BoNT/E)切割突变融合质粒pEGFP-SNAP-23ΔBoNT/E，以及鲈鱼IL-1β野生型融合表达质粒IL-1β-pEGFP和IL-1β-pEYFP。所有融合蛋白均在鲈鱼巨噬细胞内成功表达，结合ELISA实验结果发现，SNAP-23野生型的表达对IL-1β的分泌有促进作用，而Cys缺失突变体的表达则抑制IL-1β向胞外分泌。首次证实了鱼类巨噬细胞内SNAP-23蛋白在IL-1β分泌过程中的重要作用。此外通过与GFP共表达，定位了IL-1β分子在巨噬细胞内的分布，发现新合成的IL-1β分子很可能像TNFα一样经“内质网－胞质－伪足－胞外” 的分泌路径运出胞外。

评价水产动物生长性能的生理生化指标和分子生物学指标

以奥利亚罗非鱼(Oreochromis aureus)为实验对象，设计了3种不同的摄食类型，分别是鲜活饵料组、饥饿3周后饱食投喂组和人工饲料组。鲜活饵料组投喂冰冻赤子爱胜蚓，利用蚯蚓体内丰富的营养成分和活性物质，以期获得奥利亚罗非鱼良好的生长状况；饥饿后饱食组是指饥饿3周后，以人工饲料饱食投喂2周，用于研究饥饿与补偿生长获得快速生长时血液理化指标的变化情况；人工饲料组作为对照组。纯淡水条件下养殖，水温25±2℃。测定了奥利亚罗非鱼在3种摄食类型饲喂下某些血液生理生化指标变化的情况，并将指标变化情况与增重率做相关性分析，试图找出能够反映奥利亚罗非鱼生长性能的血液生理生化指标。 研究结果表明，奥利亚罗非鱼在饥饿3周后获得了补偿生长，补偿生长时的增重率和特定生长率显著高于人工饲料组(P<0.05)，高于鲜活饵料组，但差别不显著；相关性分析研究表明血清总蛋白、胆固醇、四碘甲状腺原氨酸（T4）与增重率极显著相关(P<0.01)，血红蛋白显著相关(P<0.05)，红细胞、白细胞、碱性磷酸酶高度相关(相关系数为0.580、0.551和0.557)，因此，建议血清总蛋白、胆固醇和血红蛋白可作为能够反映罗非鱼生长性能的新指标。 根据序列设计引物，PCR反应条件：变性温度：95 ℃，3 min；退火温度：57℃，20 sec；延伸温度：72℃，5 min，共36个循环，从牙鲆、黑鲪和鲈鱼中克隆出胰岛素样生长因子（IGF-Ⅰ）部分序列，首次证实了IGF-Ⅰ在3种海水鱼中的存在。 利用蛋氨酸与ZnSO4•7H2O，在pH 5.5、80℃下，反应1小时，采用蛋氨酸与硫酸锌2：1的配料比，合成出了产物蛋氨酸螯合锌，蛋氨酸螯合锌外观白色，粉状，室温下微溶于水，不溶于乙醇，并用原子吸收光谱法测定其含锌量为15%，螯合率为88.2%。

空间决策支持系统模型库系统研究

文章探讨了面向对象的模型设计、管理 .提出将模型描述、源程序、目标文件等作为一个框架的存储组织方式 ,讨论了基于 agent的模型自动生成、运行、修改方法

交易型领导的结构和作用机制研究

The Multifactor Leadership theory developed by Bass (1985) has become the new paradigm of leadership research. The empirical results of the effectiveness of transformational and transactional leadership in the literature, however, are not consentient. Researchers in China found the different structure of transformational leadership, but have not developed the transactional leadership. This study attempts to investigate three key questions in the unique Chinese socio-economic context: 1) what is the structure of transactional leadership in China? 2) What are the differences between western countries and China? And 3) what is the relationship between the transformational and transactional leadership mechanism? This study examines data collected from 3,500 participants, using Explored Factor Analysis (EFA), Confirmed Factor Analysis (CFA), Hierarchical Regression Analyses, partial correlations and other statistics methods. The major finings are listed as follows： Firstly, inductive methods was used to explore the structure of transactional leadership and the result show that transactional leadership is a four dimensions structure which includes contingent reward, contingent punishment , process control and anticipated investment. Reliability analysis, item analysis, EFA and CFA show the reliability and validity of the transactional leadership questionnaire we designed is good enough, the design of the item is effectively and properly. Contrast to other researches, anticipated investment emphasis on the leader’s recessive investment for subordinate, and this kind of transaction is quite special under the Chinese culture. While the content of the contingent reward with the contingent punishment is wider than the contingent reward in the western country, and the process control is wider than the management by exception and including goal setting and the management during the process. Secondly, hierarchical regression analyses showed that transformational and transactional leadership were significant positively related with in-role performance, extra-role performance, satisfaction and leadership effectiveness while negatively related to intention to leave. The effects of transactional and transformational leadership are different. Transactional leadership could significantly predict intention to leave controlling for transformational leadership, while transformational leadership could significantly predict in-role performance, extra-role performance, satisfaction and leadership effectiveness controlling for transactional leadership. Thirdly, the income level and the rank of subordinates are the moderators between the transformational, transactional leadership and leadership effectiveness. The leadership effectiveness of transactional leadership would decrease as the rank of subordinates increased, while the leadership effectiveness of transformational leadership would increase as the rank of subordinates increased. Transactional leadership is positively related to the effectiveness when the level of the subordinate income is low, but negatively related to the effectiveness when the level of the subordinate income is high. However the income level of the subordinate could not influence the leadership effectiveness of transformational leadership.