Thrombin Generation in Zebrafish Blood

To better understand hypercoagulability as an underlying cause for thrombosis, the leading cause of death in the Western world, new assays to study ex vivo coagulation are essential. The zebrafish is generally accepted as a good model for human hemostasis and thrombosis, as the hemostatic system proved to be similar to that in man. Their small size however, has been a hurdle for more widespread use in hemostasis related research. In this study we developed a method that enables the measurement of thrombin generation in a single drop of non-anticoagulated zebrafish blood. Pre-treatment of the fish with inhibitors of FXa and thrombin, resulted in a dose dependent diminishing of thrombin generation, demonstrating the validity of the assay. In order to establish the relationship between whole blood thrombin generation and fibrin formation, we visualized the resulting fibrin network by scanning electron microscopy. Taken together, in this study we developed a fast and reliable method to measure thrombin generation in whole blood collected from a single zebrafish. Given the similarities between coagulation pathways of zebrafish and mammals, zebrafish may be an ideal animal model to determine the effect of novel therapeutics on thrombin generation. Additionally, because of the ease with which gene functions can be silenced, zebrafish may serve as a model organism for mechanistical research in thrombosis and hemostasis.     

Repeated Blood Collection for Blood Tests in Adult Zebrafish

Repeated blood collection is one of the most common techniques performed on laboratory animals. However, a non-lethal protocol for blood collection from zebrafish has not been established. The previous methods for blood collection from zebrafish are lethal, such as lateral incision, decapitation and tail ablation. Thus we have developed a novel “repeated” blood collection method, and present here a detailed protocol outlining this procedure. This method is minimally invasive and results in a very low mortality rate (2.3%) for zebrafish, thus enabling repeated blood sampling from the same individual. The maximum volume of blood sampling is dependent on body weight of the fish. The volume for repeated blood sampling at intervals should be ≤0.4% of body weight every week or ≤1% every 2 weeks, which were evaluated by measurements of blood hemoglobin. Additionally, hemoglobin, fasting blood glucose, plasma triacylglycerol (TG) and total cholesterol levels in male and female adult zebrafish were measured. We also applied this method to investigate the dysregulation of glucose metabolism in diet-induced obesity. This blood collection method will allow many applications, including glucose and lipid metabolism and hematological studies, which will increase the use of zebrafish as a human disease model organism.     

Blood Circulation Pattern in Highly Skilled Swimmers

Echocardiography was used to study the parameters of cardiodynamics, systemic hemodynamics, left ventricular morphology and functioning, phase analysis of cardiac activity, and rheological blood properties in highly skilled expert swimmers. The relationship between different levels of blood circulation was established. A close correlation between the blood flow at rest and arterial pressure was found to contribute to the optimization of microvascular pressure and shear stress in accordance with body demands.     

Blood Flow Changes Coincide with Cellular Rearrangements during Blood Vessel Pruning in Zebrafish Embryos

After the initial formation of a highly branched vascular plexus, blood vessel pruning generates a hierarchically structured network with improved flow characteristics. We report here on the cellular events that occur during the pruning of a defined blood vessel in the eye of developing zebrafish embryos. Time-lapse imaging reveals that the connection of a new blood vessel sprout with a previously perfused multicellular endothelial tube leads to the formation of a branched, Y-shaped structure. Subsequently, endothelial cells in parts of the previously perfused branch rearrange from a multicellular into a unicellular tube, followed by blood vessel detachment. This process is accompanied by endothelial cell death. Finally, we show that differences in blood flow between neighboring vessels are important for the completion of the pruning process. Our data suggest that flow induced changes in tubular architecture ensure proper blood vessel pruning.     

外源性胰岛素对斑马鱼血糖及其转运的影响

斑马鱼（Danio rerio）在糖负荷状态下表现出持续高血糖现象。与对照组（仅腹腔注射灭菌去离子水）相比,葡萄糖组（仅腹腔注射葡萄糖）血浆胰岛素水平无显著差异,胰岛素基因表达显著上调,肝胰脏葡萄糖转运蛋白（glucose transporters,GLUTs）基因表达无显著差异,说明斑马鱼自身胰岛素分泌不足和葡萄糖转运迟缓是导致其在糖负荷状态下持续高血糖的原因。为了观察外源性胰岛素对斑马鱼血糖及其在体内转运的影响,设计低（1.25 IU/kg）、中（12.5 IU/kg）、高（125 IU/kg）3个浓度的胰岛素,分别与葡萄糖溶液（0.1 g/mL）共注射斑马鱼并观察其血糖变化。结果表明,低剂量胰岛素能有效促进斑马鱼血糖的降低,且能直观反映糖负荷后血糖的变化情况,为最适注射浓度。此外,研究显示斑马鱼血糖变化不受性别影响。在胰岛素最适注射浓度下,与葡萄糖组相比,胰岛素组（葡萄糖与胰岛素共注射）可以显著减少斑马鱼血糖恢复到正常水平的时间,进一步分析发现,斑马鱼血浆胰岛素水平增加,肝胰脏葡萄糖转运蛋白基因表达显著上调,但胰岛素基因表达却被显著抑制。综上所述,胰岛素分泌不足和葡萄糖转运迟缓是造成斑马鱼持续高血糖的原因;外源性胰岛素能够促进糖负荷状态下斑马鱼血糖的降低,但是具有反馈抑制斑马鱼肝胰脏胰岛素基因表达的作用。     

A Fall in Blood Glucose Level Precedes Meal Onset in Free-Feeding Rats

LOUIS-SYLVESTRE, J. AND J. LE MAGNEN. Chez les rats se nourrissant librement, une chute de glycémie précède chaque repas. NEUROSCI. BIOBEHAV. REV. Suppl. 1, 13–15, 1980 - L'hypothèse selon laquelle la stimulation à manger ou éveil spécifique de faim a pur origine une chute de glucèmie induite par le déficit périodique entre production hépatique et utilisation périphérique du glucose est ancienne. Cependant, jusqu'a present, les résultats de déterminations épisodiques de la glycémie, pratiquées au cours des intervalles interprandiaux, chez le rat se nourrissant ad libitum n'ont pas apporte de preuve directe. Une technique permettant une détermination continue, de longue durée, de la glycémie chez le rat se nourrissant librement a été mise au point. Son utilisation a permis de montrer que 5 à 6 minutes avant tout repas, diurne ou nocturne, se produit une chute de glycémie de 6 à 8%.     

中国水蛇雌性生殖系统的血液循环

取12条处于不同生殖期的雌性中国水蛇作血管单注射,观察其生殖系统血液循环特点。结果表明,卵巢处于静止期的个体的生殖系统循环模式与雄体相似,卵子处于迅速生长期,卵巢血管变得十分发达,布满于正在生长的卵子表面,卵子处于成熟期,卵巢血管又回复到模式状态;受精卵落入子宫中发育时,子宫血管变得十分发达,子宫壁及卵黄膜均十分薄,子宫腔中有液体浸润胚胎,提示了胚胎与母体之间可能通过这些液体进行某些物质交换。     

Imaging Transient Blood Vessel Fusion Events in Zebrafish by Correlative Volume Electron Microscopy

The study of biological processes has become increasingly reliant on obtaining high-resolution spatial and temporal data through imaging techniques. As researchers demand molecular resolution of cellular events in the context of whole organisms, correlation of non-invasive live-organism imaging with electron microscopy in complex three-dimensional samples becomes critical. The developing blood vessels of vertebrates form a highly complex network which cannot be imaged at high resolution using traditional methods. Here we show that the point of fusion between growing blood vessels of transgenic zebrafish, identified in live confocal microscopy, can subsequently be traced through the structure of the organism using Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) and Serial Block Face/Scanning Electron Microscopy (SBF/SEM). The resulting data give unprecedented microanatomical detail of the zebrafish and, for the first time, allow visualization of the ultrastructure of a time-limited biological event within the context of a whole organism.     

“血液循环”一节的教学安排

自课改以来．如何创造性地使用教材,一直是我们的一个研究课题。几年来,围绕“以学生发展为本”的目标,依据生物学课程标准,结合本地、本校的实际情况,大胆突破教材的内容编排,对所使用教材中的部分内容进行了重组、整合,从而解决了教材内容与学习时间、活动内容与活动场地等的矛盾,使课堂教学更加贴近实际,让学生在愉悦的学习氛围中主动获取新知。     

血液循环在大脑信息处理过程中的时序控制作用

自2006年以来,作者已陆续在一些杂志,网站和学术会议[1-3]上发表一些文章和证据,主要探究如下几个问题:(1)讨论各种脑电波现象的起源,大脑信息处理过程,血液循环三者之间关系.(2)提出了血液循环在大脑处理信息的过程中起到了基础时钟的作用;提出其可能的一个分子机制,其中H 浓度,O2浓度,微循环系统的工作机制,NMDA受体的H 位点是几个关键因素.(3)大脑如何感知时间的生理机制.提出时间感(或者时间流逝感)其实是一种压感,来源于当血液流经大脑的血管网的时候,造成的压感;而大脑对连续物理过程的感知则是大脑中记录情景(照片)的一个细胞群序列兴奋的信号和压感通过"捆绑效应"整合给人的感觉(即有点像放电影).在本文中,将对这些观点和证据进行回顾:文章第1节在整体上叙述所建立的脑电波模型,和"血液循环在大脑处理信息上具有时序控制作用"的观点.第2～5节将从各方面给出支持第1节中所叙述模型和观点的证据,并对一些问题进行澄清.第6～8节将该模型应用于解释一些脑电波和神经生理现象,解决目前一些问题(例如α波的起源问题,睡眠的脑电现象等).     

Numerical Models of Auto-regulation and Blood Flow in the Cerebral Circulation

A two-dimensional time-dependent computational fluid dynamics model of the Circle of Willis has been developed. To simulate, not only the peripheral resistance of the cerebrovascular tree but also its auto-regulation function, a new "active" boundary condition has been defined and developed using control theory to provide a model of the feedback mechanism. The model was then used to simulate different common abnormalities of the Circle of Willis while a pressure drop, simulating a rapid compression of the right internal carotid artery, was imposed. Test results using a simple tube compared excellently with experiment. The total time-dependent flux for each efferent artery was tabulated and showed the important relationship between geometrical variations in the Circle of Willis and the auto-regulation of blood flow by vascular vaso-dilation and contraction. From this study, it was found that the worst case seemed to be that of a missing or dysfunctional right A1 segment of the anterior cerebral artery. The use of valid physiological models of the peripheral resistance allows for more realistic models of the blood flow in the Circle whilst allowing an easy extension to 3D patient specific simulations.     

游离皮瓣移植术后血运监测的研究进展

游离皮瓣移植已经成为修复重建外科常用的修复手段之一,其成功率已经超过95%,但微循环障碍的风险仍然存在,临床上皮瓣救助率与缺血时间呈负相关,因此游离皮瓣移植术后的血运监测至关重要。作者以"皮瓣"和"监测"为检索词在PubMed数据库检索出2000年1月到2014年1月期间关于游离皮瓣移植术后监测的相关文献,纳入标准为至少5篇以上的相关报道,通过分析相关文献概述当前主要的监测方法,从临床实用性、可靠性等方面分析其优缺点。     

Alterations in Electrolyte and Trace-Element Concentrations During Simulated Extracorporeal Blood Circulation

In our studies with simulated extracorporeal blood circulation we observe damaging effects on blood cells, especially under oxygenating conditions. In order to characterize these effects we also analysed electrolyte and trace-element concentrations in plasma during and after simulated extracorporeal blood circulation. Highest resorption effects for magnesium and highest desorption effects for calcium, copper and iron are found with oxygen gas flow in the system. Membrane permeability for electrolytes seems to be induced as well. Cellular damage due only to mechanical stress within the perfusion system can be neglected.