重组细胞高产型H3N2猪流感病毒株的拯救

为拯救出一株能够在动物传代细胞中高水平复制的H3N2亚型猪流感疫苗株,利用反向遗传操作技术,将A/Goose/Dalian/3/01(H9N2)毒株的PB1、PA、NP、M、NS基因和A/PR/8/34毒株的PB2基因作为内部基因与猪流感病毒A/Swine/Henan/S4/01(H3N2)毒株的HA、NA基因进行重组,成功拯救出了具有高度细胞适应性毒株rH3N2株,该毒株接毒MDCK细胞60h后,血凝价可以达到1∶512,表明该毒株具有高度适应细胞繁殖特性,为H3N2亚型猪流感病毒细胞培养型疫苗的研制奠定了基础。     

国产膜式人工肺临床应用633例的观察和临床估价

本文总结上海市胸科医院自１９８４年９月到１９９７年１２月应用复旦大学研制的ＦＤＭＯ膜肺为６３３例病儿行心脏直视手术的实际经验。体外循环方法采用中度低温和中深度血液稀释,稀释后血球压积维持在０．１６－０．２２,心肺转流中采用高流量灌注,每分钟为２．８－３．２Ｌ／Ｍ＾２,心肺转流时间２８－３５８分钟,平均９４分钟,平均９４分钟。     

温度对黑线仓鼠能量代谢及开场行为的影响

形态、生理及行为的季节性变化是动物生存和繁殖的适应性策略。温度可能是影响啮齿动物季节性繁殖和种群波动的重要气候因子。为了验证环境温度能诱导鼠类体重、能量代谢及行为产生适应性变化的假设,并探究其中的相互关系和意义,通过不同温度处理模拟四季温度,以野生雄性黑线仓鼠(Cricetulus barabensis)为研究对象,检测其体重、日食量、静止代谢率(RMR)及开场行为的变化。结果显示:随着处理温度的下降,黑线仓鼠的体重、日食量及静止代谢率均出现显著性或极显著性地增加趋势(P0.05或P0.01);以体重为协变量的协方差分析表明,日食量和静止代谢率的增加不完全是由体重增加而造成的;高温处理降低了黑线仓鼠的爬行格数(P0.01),低温处理降低了黑线仓鼠的爬行格数和后肢站立次数(P0.01或P0.05),并产生明显的颤抖现象。以上研究结果支持环境温度能诱导鼠类体重、能量代谢及行为产生适应性变化的假设。低温同时增加黑线仓鼠能量的摄入水平和维持基本生长的能量支出水平,同时降低动物在陌生环境中的自发活动与探索行为。高温则降低黑线仓鼠能量的摄入和支出水平,同时减少自发活动与探索行为。体重、能量代谢和行为学特征的变化有利于仓鼠度过寒冷的冬季和干热的夏季,同时也与仓鼠的季节性繁殖现象相一致,因此这些特征可能是黑线仓鼠对其寒冷的冬季及干热的夏季生存环境的适应。     

干旱砾漠区露天采煤对草地植物群落特征及其稳定性影响

露天煤炭开采对区域地形地貌、水文循环和土壤植被等生态环境因子产生了较大干扰,但干旱砾漠区露天开采对周边植物群落及其稳定性的影响范围和程度仍有待深入研究,这个问题的探明才能为矿区植被恢复的精准施策提供科学依据。该文通过定量比较距露天开采区不同距离植物群落组成、多样性、生产力和稳定性变化特征,分析干旱砾漠区露天采煤对植物群落的影响范围和程度,并结合土壤性状和微气象因子的测量,探究影响干旱矿区植物群落特征的主导因子。结果表明:(1)干旱砾漠区露天开采排土场原始植被消失殆尽,开采区外围0~500 m范围内原优势物种重要值降低,伴生物种重要值增加,500 m后随着距开采区距离的增加优势植物重要值逐渐增加,至开采区800 m外植物群落组成趋于稳定;(2)开采区外围0~500 m范围内植物群落地上生物量、多样性和群落稳定性较5000 m处分别平均下降了53%、70%和37%,开采区500 m以外上述群落指标整体上变化不明显,而植被覆盖度在开采区外围不同距离并未表现出显著变化趋势;(3)土壤水分、有机质、有效氮磷钾、可溶性盐与植物群落地上生物量、多样性和稳定性呈正相关关系,而土壤pH、光照强度和地表温度与上述植物群落指标呈负相关关系。因此干旱砾漠区露天开采排土场及开采区外围0~500 m范围是矿区生态修复过程中重点考虑区域,同时研究表明干旱砾漠区植被主要分布在砾漠戈壁浅沟和季节性河道等生境相对良好的凹陷微地形条件下,综合可得出干旱砾漠区植被修复过程中可通过微地形塑造、土壤改良和乡土物种繁殖促进,以实现干旱矿区生态系统的稳定和可持续发展。     

Separation of supercoiled from open circular forms of plasmid DNA,and biological activity detection

To establish a cost-effective purification process for the large-scale production of plasmid DNA for gene therapy and DNA vaccination, a single anion-exchange chromatography (AEC) step was employed to purify supercoiled plasmid DNA (sc pDNA) from other isoforms and Escherichia coli impurities present in a clarified lysate. Two different size and conformation plasmids were used as model targets, and showed similar elution behavior in this chromatographic operation, in which sc pDNA was effectively separated from open circle plasmid DNA (oc pDNA) in a salt gradient. The process delivered high-purity pDNA of homogeneity of 95 ± 1.1% and almost undetectable levels of endotoxins, genomic DNA, RNA and protein, at a yield of 65 ± 8%. Furthermore, the transfection efficiency (29 ± 0.4%) was significantly higher than that (20 ± 0.1%) of a pDNA control. The present study confirms the possibility of using a single AEC step to purify sc pDNA from other isoforms and host contaminants present in a clarified E. coli lysate.     

Numerical investigations of transient heat transfer characteristics and vitrification tendencies in ultra-fast cell cooling processes

During freezing, cells are often damaged directly or indirectly by ice formation. Vitrification is an alternative approach to cryopreservation that avoids ice formation. The common method to achieve vitrification is to use relatively high concentrations of cryoprotectant agents (CPA) in combination with a relatively slow cooling rate. However, high concentrations of CPAs have potentially damaging toxic and/or osmotic effects on cells. Therefore, establishing methods to achieve vitrification with lower concentrations of CPAs through ultra-fast cooling rates would be advantageous in these aspects. These ultra-fast cooling rates can be realized by a cooling system with an ultra-high heat transfer coefficient (h) between the sample and coolant. The oscillating motion heat pipe (OHP), a novel cooling device utilizing the pressure change to excite the oscillation motion of the liquid plugs and vapor bubbles, can significantly increase h and may fulfill this aim. The current investigation was designed to numerically study the effects of different values of h on the transient heat transfer characteristics and vitrification tendencies of the cell suspension during the cooling processes in an ultra-thin straw (100 microm in diameter). The transient temperature distribution, the cooling rate and the volume ratio (x) of the ice quantity to the maximum crystallizable ice of the suspension were calculated. From these numerical results, it is concluded that the ultra-high h (>10(4) W/m2 K) obtained by OHPs could facilitate vitrification by efficiently decreasing x as well as the time to pass through the dangerous temperature region where the maximum ice formation happens. For comparison, OHPs can decrease both of the parameters to less than 20% of those from the widely used open pulled straw methods. Therefore, the OHP method will be a promising approach to improving vitrification tendencies of CPA solutions and could also decrease the required concentration of CPAs for vitrification, both of which are of great importance for the successful cryopreservation of cells by vitrification.     

微生物实验室管理模式初探

根据微生物学的实践教学特点, 从改革实验大纲和管理方式入手, 积极探索并初步构建了开放微生物实验室在空间场地、仪器设备、实验时间、实验内容等方面的运行与管理模式, 并在实践中收到了良好的成效。     

不同等级环境对大、小鼠及其应用的影响

目的了解不同等级环境（屏障环境和普通环境）对实验大小鼠的一般生理表现和抗应激能力以及对药物反应的影响,初步判定严格的微生物控制是否影响实验动物的人类模型作用。方法将40只SPF级Wistar大鼠和200只SPF级BALB／c小鼠分别饲养于屏障环境和普通环境内,通过以下实验对比观察不同等级环境对实验大、小鼠及其应用的影响：每周测量动物的体重增长情况;饲养35d后,测定大鼠15项血液学指标、14项血液生化指标和9个脏器的相对重量;以抗缺氧实验和游泳实验判断对小鼠体质和抗应激能力的影响;以5．氟尿嘧啶急性毒性实验和戊巴比妥钠麻醉实验测试其对药物作用的异同。结果在5周观察期内,屏障环境饲养小鼠和大鼠的体重增长均明显快于普通环境的对照动物（P〈0．05）;普通环境饲养大鼠的血液RBC、WBC、PCT、PLT、GOT、GGT、GPT、CK、TB值高于屏障环境大鼠（P〈0．05）,而CHO、LDL-C和HDL-C值低于屏障环境大鼠（P〈0．05）;不同微生物环境条件饲养小鼠的游泳耐力和抗缺氧实验结果接近,但是屏障环境组动物所获数据的变异较小;不同等级环境条件饲养小鼠的戊巴比妥钠麻醉反应和5-氟尿嘧啶中毒反应相似,而屏障环境组动物的麻醉维持时间较短,LD50较低,95％可信限狭窄。结论饲养于屏障环境和普通环境的实验大、小鼠在一般生理表现上出现部分差异,动物的抗应激能力和对药物的反应未发生显著变化;在屏障环境条件中,动物的药物反应更敏感、所获数据更稳定,因此,初步认为严格微生物控制条件下的SPF级动物可以作为生活在自然环境中的人类的模型动物。     

Opening the door to innovation

Open innovation is the new buzz, with initiatives popping up left and right. Here, we give a personal perspective on a very successful, knowledge-driven innovation initiated in an academia- industry alliance, which culminated in technology platforms that enable the generation of therapeutic antibodies with novel properties. To start, we provide a general background on open innovation in the drug development field.     

A genomic data viewer for iPad

The Integrative Genomics Viewer (IGV) for iPad, based on the popular IGV application for desktop and laptop computers, supports researchers who wish to take advantage of the mobility of today’s tablet computers to view genomic data and present findings to colleagues.