阻断5-HT1B受体对正常和帕金森病大鼠底丘脑核神经元活动的不同作用

目的观察体循环给予5-HT1B受体拮抗剂SB224289后,对正常大鼠和帕金森病(Parkinsons disease,PD)模型大鼠底丘脑核(subthalamic nucleus,STN)神经元电活动的影响。方法采用在体细胞外记录的方法,研究正常和PD大鼠STN神经元电活动的变化;比较阻断5-HT1B受体后两组大鼠STN神经元电活动的改变。结果①PD组大鼠STN神经元的平均放电频率[(7.87±1.37)Hz]明显高于正常组大鼠[(6.59±1.44)Hz,P<0.001]。两组大鼠STN神经元均表现为规则、不规则和暴发式放电3种形式。正常组大鼠具有规则、不规则和暴发式放电的神经元比例分别为33.33%、54.17%和12.50%;PD组分别为31.25%、35.42%和33.33%,PD组大鼠具有暴发式放电的神经元的百分比显著高于正常组(P<0.01)。②阻断5-HT1B受体后,正常大鼠STN神经元平均放电频率明显增高(P<0.01),具有暴发式放电的神经元数量显著增多(P<0.05)。而PD组大鼠阻断5-HT1B受体后STN神经元的放电频率和放电形式均无明显变化。结论 PD大鼠STN神经元呈过度兴奋状态;阻断5-HT1B受体可兴奋正常大鼠的STN神经元,而不影响PD大鼠的神经元活动。     

胃消化间期压力活动的相位耦合分析

目的了解胃消化间期压力活动的对角切片谱分布情况。方法应用随机信号的对角切片谱分析胃消化间期压力活动,这种方法可克服功率谱分析缺少相位信息的缺陷,同时保留高阶谱分析包含相位信息的优良特性。结果胃消化间期移行性复合运动(MMC)Ⅲ期压力活动表现明显的平方相位自耦合结构,而Ⅰ期的压力活动几乎没有相位耦合。结论胃消化间期压力活动可能存在着非线性平方相位耦合关系,并从细胞分子水平上的胃肌电活动(GMA)对这种耦合现象发生的机理作了讨论。     

乙型肝炎病毒全X基因对HEK293细胞功能的影响

目的克隆并构建乙型肝炎病毒(HBV)全X基因表达载体,探讨HBV全X基因对HEK293细胞功能的影响,并比较HBV全X基因与X基因对细胞影响的差异。方法应用基因克隆的方法从慢性HBV感染者血清中获得HBV全X及X基因,构建带有HBV全X基因与X基因的真核表达载体以及带有绿色荧光基团GFP的重组质粒;细胞转染技术将其分别转入HEK293细胞,RT-PCR及Western-blot技术验证HBV全X基因及X基因在细胞内的表达;荧光显微镜下观察其在细胞内的定位情况;MTT法检测细胞增殖率的变化;Annexin V流式细胞分析技术检测其对细胞凋亡率的影响;TRAP实时荧光定量PCR法检测各组细胞端粒酶活性的变化。结果成功构建带有HBV全X基因的重组质粒且目的基因能在HEK293细胞内高表达;表达的HBV全X蛋白主要分布在胞核,而X蛋白在胞质、胞核均有分布;转染HBV全X和X基因后细胞增殖率分别为(0.826±0.002)%和(0.805±0.013)%,显著高于转染空质粒(0.691±0.035)%和未转染对照组(0.681±0.018)%,差异具有统计学意义(P<0.05);转染HBV全X和X基因后细胞凋亡率分别为(25.25±3.02)%及(21.34±2.16)%,显著高于转染空质粒(13.49±1.49)%及未转染细胞对照组(11.37±1.58)%,差异具有统计学意义(P<0.05);转染全X及X基因细胞的端粒酶活性亦显著高于对照组,差异有统计学意义(P<0.05);全X转染组细胞的增殖率、凋亡率及端粒酶活性均高于X转染组,但差异均无统计学意义(P>0.05)。结论 HBV全X基因具有促细胞增殖、凋亡以及影响细胞端粒酶活性的作用,其可能通过不同于HBV X基因的细胞内途径影响细胞的生物学功能,从而在HBV相关的原发性肝细胞癌发生发展中起作用。     

交通拥堵与温室气体排放

缓解拥堵项目可减少温室气体排放,但基于单个变量(如仅基于出行距离)的CO2排放预测并不精确.为了准确预测缓堵项目减少的CO2排放量,利用车辆运行检测技术、车辆活动数据库及根据车辆类型设计的排放模型,通过分析速度与排放的关系绘制速度-排放曲线,并结合交通运行检测数据,评价缓堵策略、速度管理策略、交通平滑策略等交通运行管理...     

艾里莫芬烷倍半萜抗白血病定量构效关系的量子化学研究

采用Gaussian03程序用密度泛函理论计算了15种艾里莫芬烷倍半萜的量子化学参数,研究了这类化合物抗白血病生物活性与量子化学参数间的关系,发现化合物的生物活性与LUMO和LUMO+1轨道的能量以及偶极有较好的线性关系,LUMO和LUMO+1轨道的能量越低,偶极越大,则生物活性越好。而这类化合物的LUMO轨道主要是由内酯环或呋喃环组成的π分子轨道,内酯环是化合物的生物活性区域。研究结果为此类化合物的结构修饰和开发研究提供了参考依据。     

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基于支持向量机理论对出行链活动类型的识别方法进行了研究. 首先对居民出行的时间序列位置信息做数据预处理,提取出行链的出行过程和活动地点信息,并结合地理信息系统（GIS）提取活动的备选类型;然后从出行链和活动的时间和空间因素提取活动类型识别的特征,形成特征向量作为分类器的输入,并建立基于支持向量机的两两分类器,采用分类器投票的方法从备选集中选择活动的类型;最后利用模拟数据和交叉验证的方法对两两分类器进行训练检验,分别从高斯径向机核函数和多层感知器核函数的角度分析活动类型识别率. 结果表明：在两两分类中,高斯径向机核函数的最高识别率为99%,最低识别率为62%;多层感知器核函数的最高识别率为97%,最低识别率为54%.     

基于支持向量机的出行链活动类型识别研究

基于支持向量机理论对出行链活动类型的识别方法进行了研究. 首先对居民出行的时间序列位置信息做数据预处理,提取出行链的出行过程和活动地点信息,并结合地理信息系统（GIS）提取活动的备选类型;然后从出行链和活动的时间和空间因素提取活动类型识别的特征,形成特征向量作为分类器的输入,并建立基于支持向量机的两两分类器,采用分类器投票的方法从备选集中选择活动的类型;最后利用模拟数据和交叉验证的方法对两两分类器进行训练检验,分别从高斯径向机核函数和多层感知器核函数的角度分析活动类型识别率. 结果表明：在两两分类中,高斯径向机核函数的最高识别率为99%,最低识别率为62%;多层感知器核函数的最高识别率为97%,最低识别率为54%.     

语言交际能力培养在航海英语口语教学中的实施

航海英语口语教学的现状偏离了培养学生语言交际能力的宗旨.本文从阐述语言交际能力的定义入手,分析培养语言交际能力的途径和手段,并结合航海英语的特点,提出在课堂环境中,培养语言交际能力的实施方法.     

Preliminary study on Salvia miltiorrhiza bung endophytic fungus

Objective To select the strains which can produce tanshinone ⅡA like its host plant Salvia miltiorrhiza bung. Methods A total of 50 strains of endophytic fungi were isolated from healthy, living and symptomless tissues of Salvia miltiorrhiza bung, among which 29 strains were obtained from the root, 14 from the stem, 3 from the leaf, 3 from the flower and 1 from the seed. Their antimicrobial activities against nine different bacteria, including both Gram-negative and Gram-positive bacteria, were measured by Oxford plate agar diffusion bioassay. Results Our data showed that all but four strains had significant antibacterial activities on at least one indicator bacterium to some extent, and five strains (DR1, DR4, DR16, DR18 and DF2) manifested quite prominent antibacterial activities against certain pathogenic bacteria. In some degree, it might indicate that this endophytic fungus isolated from the tissues of Salvia miltiorrhiza bung has a potential value as a natural antibacterial medicine as well. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were carried out to test selected strains, both inside and outside of the cell to see if any strain can produce tanshinone ⅡA. The result showed that extracts from three strains, labeled as DR12 (outside cell), DR21 (inside cell) and DF3 (inside cell), had a component with the same Rf value in TLC assay as that of authentic tanshinone ⅡA. The extract from DR12 (outside cell) and DR21 (inside cell) had a peak at retention time identical to that of authentic tanshinone ⅡA in HPLC. Conclusion The fungi appear to produce the bioactive compound tanshinone ⅡA, and they could be used to produce tanshinone ⅡA by fermentation. It provides a new way to synthesize this natural medicine.     

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.