DH10B菌株高效电转化条件探究

以pUC19、pECBAC1、pCLD04541DNA以及3个不同大小的BACDNA为材料,研究了E.coli DH10B菌株在5个不同脉冲电场下的转化效率。研究发现,随着DNA片段大小的增加,最高转化效率和最适场强迅速减小。利用DH10B细胞转化pUC19 DNA的最适场强是21kV/cm,而190kb BAC DNA仅为13kV/cm;在最适场强下,40kb BAC DNA的转化效率约是190kb BAC DNA的50倍。通过大量数据绘制了不同因素影响下转化效率的变化曲线,优化了E.coli DH10B菌株电转化条件,为质粒的重组转化以及大片段基因组文库的构建奠定了基础。     

脉冲电场对真皮成纤维细胞生长和膜流动性的影响

采用MTT比色分析法检测脉冲电场(f=50Hz,t=20μs,Epp=1V/m),对真皮成纤维细胞增殖的影响,结果表明,电场作用1分钟和5分钟,均产生促进细胞增殖的效果(P<0.05和P<0.001),而较长时间(t≥10分钟)的电场作用则显著抑制细胞的正常增殖(P<0.001)。 采用荧光偏振法研究了脉冲电场对真皮成纤维细胞膜流动性的影响。结果表明,电场作用5分钟后即刻引起膜流动性的显著增加。而电场作用45分钟后,需经过一段时间的温育(30 分钟),才表现出膜流动性的降低。说明不同作用时间的电场可以造成膜流动性的增加或降低,而且对膜流动性的影响是可逆的。我们认为,脉冲电场所引起的膜流动性改变是细胞分裂能力发生变化的原因之一。     

脱落酸、多效唑和油菜素内酯对低温期间香蕉过氧化物酶和电导率的影响

在人工气候箱中模拟寒潮对香蕉幼苗造成的低温冷害 ,研究脱落酸、多效唑和油菜素内酯对低温胁迫期和低温胁迫恢复期香蕉幼苗叶片过氧化物酶和相对电导率的影响。试验表明 ,叶面喷施适量脱落酸、多效唑和油菜素内酯能够提高低温胁迫期和恢复期香蕉叶片的过氧化物酶活性并且降低叶片相对电导率。其最适浓度脱落酸为 2 0～ 2 5mg/ L;多效唑为 15～ 30 m g/ L;油菜素内酯为 0 .9～ 1.5m g/ L。     

急性脊髓损伤后大鼠电刺激运动诱发电位的变化

目的：比较不同程度脊髓损伤（SCI）与运动诱发电位（MEP）变化之间的关系,探索MEP检查在SCI早期诊断及预后中的价值。方法：27只雄性SD大鼠以改良Allen‘s打击法致伤T8－T9脊髓,按打击冲量随机分为空白对照组（n=5),SCI A组（50gcf,n=8),SCI B组（70gcf,n=8)和SCI C组（100gcf,n=6),采用单极经皮层电刺激,分别于损伤前、伤后即刻、15min、30min、1h、3h和6h连续观察scMEP变化,并计算脊髓出血坏死区域占脊髓横截面积的比率。结果：对照组MEP无显著改变,SCI A组和SCI B组动物MEP早成份波幅立即减低或消失,以后有所恢复,晚成份波消失后未再出现。SCI C组动物除2只大鼠SCI后MEP仍有所恢复外,其余动物再未出现MEP波。脊髓损伤随打击冲量增大而增加,与伤后1h scMEP最大波幅呈显著相关（r=-0.821)。结论：SCI后scMEP的变化程度与打击冲量和脊髓病理损伤面积相关,提示scMEP可以作为一种脊髓功能检测的客观指标。     

Directional sensitivity of tuberous electroreceptors: polarity preferences and frequency tuning

This paper examines the directionality of tuberous electroreceptor responses and relates them to a polarity bias seen for passive electrolocation by electric fish (Hypopomus). We recorded from Burst Duration Coders (BDCs) while stimulating with 1 kHz single period sine waves with electric fields oriented horizontally in different directions. Electroreceptors have figure-8 directional sensitivity profiles with two, usually unequal lobes of sensitivity separated by 180°. For most units the larger lobe points inward, while for a few, the lobes are symmetrical or the larger lobe points outward. The differences correlate with differences in frequency tuning of the receptors. We can alter, and even reverse, the directional asymmetry of a single unit by changing the frequency of the stimulus. Two general response profiles result, with two corresponding classes of tuning curves. The degree of asymmetry varies with position on the body surface. The asymmetries and the effects of stimulus frequency and of tuning can be modeled with a linear/non-linear/linear cascade filter. The behavioral preference for approaching the head end ( + ) of an electrode is difficult to understand in light of the asymmetry of responses we report for amplitude-coding BDCs but can be understood by reference to the time-coding Pulse Marker (PM) receptors.Abbreviations BDC Burst Duration Coder - EOD electric organ discharge - nALL anterior lateral line nerve - PM Pulse Marker     

神经球方法在神经生物学中的应用及其局限性

神经球是神经干细胞在体外扩增培养过程中的一般表现形式。目前,神经球方法(neurosphere assay,NSA)已在神经干细胞性质、神经发育模型研究和作为分子载体用于神经系统疾病治疗等方面得到了广泛应用,但因神经球固有的异质性、自发融合等特性限制了其应用。本文就神经球方法在神经生物学研究中的应用及其局限性做一综述。     

富氢盐水对高压电烧伤大鼠微循环内红细胞流速及组织病理损伤的影响

目的:探讨富氢盐水对高压电烧伤大鼠微循环内红细胞流速及组织病理损伤的影响。方法:80只大鼠随机分为四组,分别为烧伤组、治疗组、对照组和阳性对照组,治疗组烧伤后给予富氢盐水治疗,阳性对照组给予罂粟碱溶液治疗。每组分成三个亚组用于血气分析,肺组织形态学观察和红细胞流动速度检测。结果:治疗组和阳性对照组大鼠肺组织湿干重比均显著低于烧伤组(P0.05),治疗组和阳性对照组大鼠肺组织湿干重比差异不显著(t=0.585,P0.05)。治疗组和阳性对照组大鼠生存时间均显著高于烧伤组(P0.05),治疗组和阳性对照组大鼠生存时间差异不显著(t=0.448,P0.05)。治疗组和阳性对照组氧分压均显著高于烧伤组(P0.05);治疗组和阳性对照组氧分压差异不显著(t=0.424,P0.05)。治疗组和阳性对照组二氧化碳分压均显著低于烧伤组(P0.05);治疗组和阳性对照组二氧化碳分压差异不显著(t=1.285,P0.05)。治疗组和阳性对照组大鼠红细胞流动速度分压均显著高于烧伤组(P0.05);治疗组和阳性对照组之间大鼠红细胞流动速度差异不显著(P0.05)。结论:富氢盐水能够促进烧伤大鼠微循环内红细胞的流动,改善微循环,恢复受损肺组织,在高压电烧伤患者的治疗方面具有较好的应用前景。     

基于无人机的冬小麦拔节期表层土壤有机质含量遥感反演

快速监测大面积分布的盐渍化麦田土壤有机质含量,可为推进盐渍土改良和促进碳循环研究提供数据支撑。通过野外采样与获取无人机遥感影像,分别基于裸土和植被情况,采用多元线性回归(MLR)、偏最小二乘回归(PLSR)和支持向量机回归(SVR)3种方法,建立区域有机质含量遥感模型,并进行检验和对比,确定最优的土壤有机质含量反演模型;最后基于最优模型进行研究区表层土壤有机质的反演,并与插值结果进行比较。结果表明: 经5×5的中值滤波处理后的光谱与土壤表层有机质对应最优;3种模型中,SVR模型的预测精度最高,PLSR次之,MLR效果最差。对比两种变量的建模效果,基于植被的SVR建模效果最好,其建模决定系数(R²)、均方根误差(RMSE)分别为0.89、0.20,验证R²、RMSE分别为0.82、0.24;基于裸土的建模效果不理想,最优的也是SVR模型,其建模R²、RMSE分别为0.63、0.26,验证R²、RMSE分别为0.61、0.25。根据最优模型反演得到该区域有机质含量为17.51～22.53 g·kg^-1,平均值为19.51 g·kg^-1,与实地调查结果较为一致;插值结果与反演结果相比,精度受到限制。综上,基于无人机多光谱可以对盐渍土冬小麦拔节期土壤有机质含量进行快速、大范围精准估测。     

A simplified model for spatially differentiated impact assessment of air emissions

We developed a simplified emission dispersion and exposure-assessment model designed to reflect the site-specific health impacts of air pollution in life-cycle impact assessment (LCIA). We proposed an EXposure Per Emission Coefficient (EXPEC), a dimensionless parameter representing the relative amount of pollutant inhaled per emission. EXPEC values were calculated for two typical source categories - electric power plants and road vehicles — on a concentric circle model. The EXPEC values were significantly different for different locations and source categories. We examined the application of EXPEC in a case study that compared the effects of emissions from electric and gasoline-engine vehicles. EXPEC is a useful tool for assessing spatially differentiated potential impacts.     

General and transport properties of hypotonic and isotonic preparations of resealed erythrocyte ghosts

Summary Resealed human erythrocyte ghosts are regarded as valuable tools for the study of membrane properties. In order to investigate to what extent preparation procedures affect the yield of ghosts, their general properties, and their permeability, ghosts prepared by lysis at low (hypotonic media) and high (isotonic media) ionic strength were compared with each other and with native erythrocytes. For isotonic lysis, cells were either subjected to dielectric breakdown or suspended in isotonic NH₄Cl solutions. In spite of very different characteristics of the lysis and the resealing process in the three types of preparations, the resulting ghosts do not differ in a number of features except for somewhat varying yields and for properties resulting from the mode of lysis.Specific transport properties, as characterized by the mediated fluxes ofm-erythritol,l-arabinose,l-lactate, and sulfate, proved to be unaltered with a few unsystematic exceptions. The simple nonmediated fluxes of all these permeants, as measured in the presence of inhibitors, however, were enhanced between 1.5- and 4-fold, indicating a somewhat increased ground permeability (of the lipid domain) in all ghost membranes.