模拟微重力条件下心肌细胞的体外三维固定化培养

观察心肌细胞体外培养形成三维(3D)组织结构的能力和过程及心肌细胞在模拟微重力状态下的3D固定化培养效果。应用酶消化法从新生的乳鼠心室肌组织获取心肌细胞,以Cytodex3为心肌细胞的3D固定化培养载体,将心肌细胞固定化培养于Spinnerflask中,用扫描电镜观察心肌细胞体外培养形成的3D组织结构;以心肌细胞的代谢效率和细胞搏动强度为观察指标,比较心肌细胞在Spinnerflask及HARV(highaspectratevessel)生物反应器中3D固定化培养的差异。结果显示,心肌细胞不仅能贴附于Cytodex3上生长,且形成了具有同步自律收缩的3D组织样结构;心肌细胞在两种不同培养体系中的细胞接种效率和细胞形态没有明显差异,培养于HARV中的心肌细胞的代谢效率和细胞搏动强度均明显高于Spinnerflask培养体系。体外培养的乳鼠心肌细胞具有形成同步自律收缩的3D组织结构的能力;模拟微重力的培养环境有利于改善心肌细胞3D组织样培养物的代谢和功能 。     

培养新生大鼠心肌细胞的电信号传导：多电极记录研究

利用多电极阵列同步记录技术对培养的新生大鼠单层心肌细胞的电活动进行胞外记录,观察心肌细胞在自发搏动和电刺激情况下信号在细胞间的传导模式。通过对记录信号的处理和分析,能获得诸如起搏细胞的数量和位置、动作电位的传导速度和途径以及不同起搏细胞间的相互影响等信息。研究还发现,心肌细胞阈下刺激会影响细胞的搏动和信号传导。     

原代单层培养的大鼠搏动心肌细胞内电位的记录方法

自发性节律搏动是原代单层培养心肌细胞的一个重要特征。培养心肌细胞的平均直径较小,自发性节律搏动频率较快。用玻璃微电极记录培养心肌细胞的电位,穿刺成功率较低,且难以稳定在细胞内。我们采用加硅涂层的玻璃微电极方法能在单层搏动培养心肌细胞内稳定地记录1h以上。 所用玻璃微电极是由内含玻璃纤维,外径为1mm的毛细管加热后,经两次拉曳,得到颈长15mm,尖端直径<0.1μm,长度为2～3μm的微电极。并在管内灌入3mol/L KCl溶液,直流电阻为30～50MΩ。在微电极灌注电解质液后,将其尖端浸在含2％二甲基二氯硅烷的四氯化碳溶液内硅化数秒钟。     

微重力条件下心肌细胞在聚羟基乙酸纤维支架上的三维固定化培养

以 1d龄Wistar乳鼠的心室肌组织为心肌细胞的来源 ,采用胰蛋白酶消化及细胞差速贴壁分离心肌细胞 ,以未经修饰和经鼠尾胶原溶液浸泡修饰的聚羟基乙酸 (polyglycolicacid ,PGA)纤维支架作为心肌细胞体外三维 (3D)固定化培养的支架 ,比较心肌细胞在静置培养体系及微重力培养体系下的生长、形态和收缩状况。心肌细胞在未经处理的PGA纤维支架 3D固定化培养时 ,心肌细胞在其上的分布不均匀 ,大部分心肌相互连接形成球状聚集体 ;PGA纤维支架经鼠尾胶原溶液浸泡处理后 ,心肌细胞在其上的分布较为均匀 ,细胞形态多呈梭形或不规则状 ,心肌细胞自律性搏动的幅度加大。在模拟微重力培养条件下 ,心肌细胞在经鼠尾胶原溶液浸泡修饰的PGA纤维支架上的分布更为均匀 ,心肌细胞形成具有自律性同步收缩特性、面积约为 15mm2 的类组织样 3D结构     

白细胞介素-2对心脏节律的作用及其机制的研究

目的：探讨白细胞介素－2（1L－2）对心脏节律作用及其可能机制。方法：采用体外培养乳鼠心肌细胞模型和离体人鼠灌流心脏模型,观察培养的心肌细胞搏动频率和离体心脏心率及节律。结果：①2.5-200u/ml的IL－2呈浓度依赖性地降低心肌细胞的搏动频率。②50u/ml的IL-2明显增加离体心脏心率和室性早搏个数。③propranolol预处理可取消50u/mlIL-2的离体心脏作用。④热失活IL－2对培养的心肌细胞搏动频率和离体心脏心率和心律都无显著作用。结论：IL－2可直接制培养心肌自律性,其对离体心脏的正性变时和致心律失常作用可能由内源性作茶酚胺介导。     

β3受体激动剂对培养的大鼠心肌细胞搏动频率和细胞内环-磷酸腺苷水平的影响

目的：观察β3受体激动剂(BRL-37344)对培养的大鼠心肌细胞搏动频率和细胞内环-磷酸腺苷(cAMP)水平的影响,以探讨β3受体在心肌细胞中的作用。方法：分离培养乳鼠心肌细胞,随机分为八组：对照组、ISO组、Nadolol ISO组、BRL组、PTX BRL组、L-NAME BRL、Nadolol BRL组和Buprandol BRL组,观察心肌细胞搏动频率,并应用酶联免疫方法测定cAMP含量,逆转录多聚酶链反应(RT-PCR)方法测β3受体mRNA表达。结果：ISO(非选择性β受体激动剂)可显著增加心肌细胞搏动频率和升高cAMP水平,这种作用可被Nadolol(为β1,β2受体抑制剂)阻断。BRL-37344可显著降低心肌细胞搏动频率和cAMP含量,这种作用可被PTX(Gi蛋白抑制剂)和Bupranolol(非选择性β受体阻滞荆)完全阻断,同时可被L-NAME(一氧化氮合酶抑制剂)部分阻断,不受Badolol影响。RT-PCR方法测出心肌细胞中有β3受体mRNA表达。结论：心肌细胞中存在β3受体,它在心肌表现为负性变力作用,β3受体的效应不受β1,β2受体抑制剂影响。心脏β3受体信号途径中可能有Gi蛋白的参与,并且经过一氧化氮合酶途径发挥其作用。     

双光电描记体外培养心肌细胞搏动的方法

本文阐述一种新的体外培养心肌细胞搏动双光电描记系统,能同时描记出同一视野不同细胞(簇)的搏动变化,同步分析不同生理或病理状态的心肌细胞在各种因素刺激下的各自搏动特征,准确测定出细胞搏动发生同步的时间,研究冲动在心肌细胞间传导过程及细胞之间的互相关系,并有简便、效率高、节省细胞标本之优点。     

组胺对培养新生大鼠心肌细胞自发性搏动及动作电位的影响

以培养大鼠乳鼠心肌细胞为模型,观察组胺对培养心肌细胞自发性搏动频率及动作电位的影响。结果表明：组胺０．１－１０ｕｍｏｌ／Ｌ可以引起剂量依赖性的心肌细胞搏动频率的增快,而且这种效应呈时间依赖性。组胺１０ｕｍｏｌ／Ｌ可以使心肌细胞动作电位的幅度（ＡＰＡ）,最大上升速率（Ｖｍａｘ）及超射（ＯＳ）明显增加,动作电位持续时间（ＡＰＤ５０和ＡＰＤ９０）明显延长,窦性周长（ＳＣＬ）明显缩短。以上结果提示,组胺致     

以胶原膜为支架的心肌细胞体外三维培养

将从新生乳鼠心室肌组织获取的心肌细胞接种于鼠尾胶原膜三维支架和组织培养板,以细胞形态、细胞搏动、葡萄糖比消耗率(q_glu)、乳酸比产率(q_lac)、乳酸转化率(Y_lac/glu)、肌酸激酶及乳酸脱氢酶的活力为观察指标,比较心肌细胞在鼠尾胶原膜中三维(3D)培养和组织培养板中二维(2D)培养的差异。培养于鼠尾胶原膜的乳鼠心肌细胞在第5天形成闰盘连接,形成面积约为80mm³、肉眼可见自律性同步收缩的心肌细胞3D培养物。3D培养体系中乳鼠心肌细胞的q_glu、q_lac和Y_lac/glu的均值分别为7.37 μmol/10.6cells/d、2.92 μmol/106cells/d和0.38 μmol/μmol;2D培养体系中乳鼠心肌细胞的q_glu、q_lac和Y_lac/glu的均值分别为7.59 μmol/10.6cells/d、3.83 μmol/10.6cells/d和 0.51 μmol/μmol。两种培养体系中乳鼠心肌细胞的肌酸激酶及乳酸脱氢酶的活力无明显差别。实验结果表明：培养于鼠尾胶原膜的心肌细胞保持正常心肌细胞的代谢活力和收缩功能。     

铬对增强心肌细胞抗病毒作用的研究

本文研究了Coxsackie B_2病毒(XOX B_2V)对人及鼠心肌细胞的感染性以及三氯化铬对心肌细胞生长与对病毒感染佳的影响。实验结果证明人心肌细胞对COX B_2 V感染的敏感性比鼠心肌细胞高。三氯化铬能促进人与鼠心肌细胞的生长,促进鼠心肌细胞的搏动及搏动频率加快,并随着培养时间的增加,细胞生长代谢旺盛,细胞培养液中乳酸脱氢酶(LDH)的含量升高。用含铬培养液培养的人心肌细胞对病毒感染的敏感性有所降低,人心肌细胞培养液中LDH的含量随病毒对细胞损伤的程度而升高。     

A force transducer to measure individual finger loads during activities of daily living.

A new six-degree-of-freedom force transducer has been manufactured, with the sensitivity to measure forces in the range +/-100 N and moments of up to +/-5 Nm. The transducer incorporates two mechanical components: shear forces and bending moments are measured via a strain-gauged tubular section whilst axial forces are transmitted to a cantilevered load cell. Both components are instrumented with 350 ohms strain gauge full bridge circuits and are temperature compensated. After calibration, measurement errors are less than +/-0.3 N for direct forces and +/-0.03 Nm for applied moments. In order to measure sub-maximal finger loads during activities of daily living, the transducer has been incorporated into several housings representing objects in domestic use: a jar, a tap, a key in a lock and a jug kettle.     

自由活动大鼠睡眠状态和呼吸节律的同步监测方法

目的:设计一种睡眠呼吸暂停综合征动物模型的监测方法.方法:通过多导生理仪同步监测自由活动大鼠6 h生理睡眠时间的皮层脑电、颈部肌电,以及呼吸波形.并用人机交互的方法进行数据处理.结果:监测信号稳定、清晰易辨,根据记录到的信号可进行不同清醒-睡眠状态分期,并判断大鼠的呼吸暂停情况.结论:该法操作简便,是小动物睡眠和呼吸监测的可靠方法.     

微机控制的穿棱箱双向主动回避反应实验系统

目的：研究动物的学习与记忆功能。方法：研制一种微机控制的穿梭箱双向主动回避反应实验系统,自动分析和记录实验结果。结果：实验结果客观、准确。结论：本系统自动化程度高,操作简便,能满足实验需求     

A method to record circadian plant movements, with application to Oxalis leaf rhythms

A transducer was developed to record the circadian movement of the individual leaflets in Oxalis regnellii Mig. The method can easily be adapted to measure other kinds of plant movements as well. It is based on the detection of the shadow each leaflet casts on the small side of a specially formed Perspex plate. The light is guided through the Perspex and collected by a phototransistor, which provides an electrical signal that is proportional to the light intensity falling onto it. The output signal can be made a linear function of the leaf angle. This equipment was used in experiments to study the coupling between the 3 leaflets in Oxalis . Pulses of 4 h of red light were given to one of the leaflets, the two others were shielded from the light. A phase response curve was determined for each leaflet, but there was no significant difference in the phase response between the 3 leaflets. Experiments were also made in which the 3 leaflets were separated physically by cuts along the petiole between the pulvini. In this case ultradian oscillations were observed.     

利用连续光声成像监测动物脑部血液动力学变化

基于B型扫描超声诊断仪和线性阵列探测器的快速光声成像系统被搭建,并用于脑部血液动力学的监测研究。实验中,小鼠脑部皮层的全景血管网络分布图像通过光声探测器双扫描采集接收的方式被重建出来。然后,针对小鼠的脑中动脉,利用阵列电子扫描采集方式,光声成像实现了局部连续地监测由静脉注射生理盐水和吲哚菁绿引起的脑部血液动力学反应,成功地利用光声图像记录其光吸收的改变。实验展示了光声快速成像获取脑部功能信号的应用潜力。     

A streptavidin surface on planar glass substrates for the detection of biomolecular interaction

Based on the requirements of biomolecular interaction analysis on direct optical transducers, a streptavidin surface is examined. A general protocol was developed allowing the immobilization of biotinylated compounds using the rife biotin-streptavidin system. This type of surface modification can be applied to all biosensors using glass surfaces as sensor devices. Reflectometric interference spectroscopy (RIfS), a label-free, direct optical method was used to demonstrate the quality of the transducer surfaces. The surface modification is based on an aminofunctionalized polyethylene glycol layer covalently bound to the silica surface of the transducer and shows very little nonspecific binding. Biotin molecules can be easily coupled on such layers. Streptavidin followed by a biotinylated estrone derivative was immobilized by incubation of the biotinylated transducer surface. For the streptavidin layer we obtained interference signals corresponding to a protein monolayer. Finally, using a surface prepared as described above, biomolecular interaction experiments with an antibody against estrone were carried out to show the quality of the transducer surface. With RIfS all of the affinity-based surface modifications can be detected online and time resolved.     

Three-dimensional interdigitated electrode array as a transducer for label-free biosensors

A new transducer for biosensor applications has been developed based on a three-dimensional interdigitated electrode array (IDEA) with electrode digits separated by an insulating barrier. Binding of molecules to a chemically modified surface of the transducer induces important changes in conductivity between the electrodes. Three-dimensional sensor shows considerable improvement compared with a standard planar IDEA design. The potential of the developed device as a sensor transducer to detect immunochemical and enzymatic reactions, as well as DNA hybridization events is demonstrated. The immunosensor allows direct detection of the antibiotic sulfapyridine and shows the IC(50) parameter value of 5.6 microgL(-1) in a buffer. Immunochemical determination occurs under competitive configurations and without the use of any label. Each modified sensor is of a single use. Nevertheless, biochemical reagents can be easily cleaned off the sensor surface for its reuse. Layer-by-layer method of used to deposit polyethyleneimine and glucose oxidase showed that the sensor is also highly effective for detecting single and multilayered molecular assemblies.     

竹亚科簕竹属吊罗坭竹繁殖器官的补充描述

《中国植物志》第九卷第一分册及其它的相关文献均无关于竹亚科簕竹属吊罗坭竹花的描述或记录,本文作者采集制作了该竹子繁殖器官标本,并分别用中文和拉丁文对其进行了描述,其主要特征：花枝长70~170 cm;花序为总状花序或简单的圆锥花序;花序轴粗,节明显;小穗含4~5朵小花;小穗无柄;小穗轴节间无毛,呈棒状;颖2,膜质;外稃长或等于内稃;鳞被2;雄蕊6;子房卵形;柱头3,呈毛刷状;颖果未见。花期2~3月。花标本采自厦门市园林植物园。     

Emerging array-based technologies in proteomics

Microarray format is the method of choice for high-throughput hybridization of nucleic acids. Lately, microarray strategy has been applied to broad studies of interactions of proteins with other molecules. The diversity of these interactions and variations of the properties of proteins present new challenges to microchip technology. Significant achievements have been reported. In particular, three-dimensional (gel-based) chips can be manufactured with limited resources and offer flexibility, capacity, homogeneous solvent environment and the ability to record processes in real time and in variable conditions.     

The Vibratome-Ralph knife combination: a useful tool for immunohistochemistry

A modified immunohistochemical technique is described for the improved detection of antigens. The method involves the use of the Vibratome combined with Ralph knives, which are easily manufactured with an LKB 2078 Histoknifemaker. The sectioning procedure was used with the peroxidase-antiperoxidase method of Sternberger and his collaborators for demonstrating growth hormone release-inhibiting hormone (somatostatin) and noradrenaline N-methyltransferase in neurones in the central nervous system of the rat. The morphological preservation of the tissue was good and cytological details were easily seen, especially in the thin sections (5-10 microns thick).