这项研究是由德国联邦Ministery教育与研究部（BMBF）资助的"虚拟肝络"的支持。我要感谢弗兰克·舒伯特和刘若英贡佩尔特从耶拿大学医院为他们生产的视频和创建动画和伊莎贝尔JANK录制音频帮助媒体中心。

住房和进行的所有程序均按照德国动物福利立法。 1，传感器校准（按照制造商的说明进行传感器校准） 1.1）米勒导管校准。预浸泡在导管的尖端在无菌水或盐水的前30分钟，以平衡（归零）和校准。 <ol><li>在米勒传感器连接到桥式放大器的millar1通道，插入米勒传感器尖端插入水柱。 </li><li>设置水柱值:0水柱2 O在数据分析软件窗口中，选择桥放大和零吧。基线值0 水柱可以设置。 </li><li>设置水柱值到20水柱2 O运行数据分析软件窗口的进展，并停止。选择在桥的窗口"单位"放大，2 O相应地设置为0和20水柱基线。调整"单元"，以水柱&lt;子&gt; 2 O。 </li><li>校准millar2用于测量CAP中相同的方式（设置两个基准线90和110 水柱 ）。 </li></ol> 1.2），血流量探头校准<ol><li>将探头置于去离子水。连接探头与跨音速流动探头系统。 </li><li>在数据分析软件窗口中，选择输入放大到零流量探头。调整单元。 </li><li>按下按钮"测试信道"来收集信号:如果信号具有3-4条，这意味着该信号是好的。如果一个好的信号获取，程序可以继续。 </li><li>按下按钮，以"零通道"和规模频道看到的值是否已经校准与否。 </li><li>按下按钮"测量通道"后测量。 </li></ol> 2，准备好鼠标的手术过程<ol><li>鼠标放置在一个感应室和麻醉小鼠，用2％异氟醚和0.3ml /分钟的氧气。该操作可以在鼠标的脚趾捏撤回反射不存在下进行。 </li><li>剃手术区域，其包括左颈部和腹部的毛。 </li><li>把鼠标放在手术台上，并用胶带固定。使用对眼睛的兽医药膏，以防在操作期间干燥。 </li><li>将脖子下纱布垫在脖子上的操作领域的最佳曝光。 </li><li>消毒操作领域，并把消毒纱布覆盖鼠标只留下手术视野打开。 </li></ol> 3，重要参数的测量<ol><li>将心针进入皮下小鼠的爪子。 </li><li>将呼吸传感器下的小鼠的背部。 </li><li>将温度探头插入鼠标的直肠。 </li><li>记录温度，心电图和鼠标在数据分析软件的呼吸频率。 </li></ol> 4，颈部操作局长ystemic心血管监测 4.1）血管夹层<ol><li>确定中线颈，中间点锁骨，下颌骨的角度。 </li><li>使从下颌到锁骨的中间点是0.5厘米到中线的左边的角2cm的纵向切口。 </li><li>解剖颌下腺，翻过来，并用生理盐水浸泡的纱布覆盖。 </li><li>找出颈内静脉，解剖它，把静脉结扎后并固定在三个6-0丝线缝合。 </li><li>确定胸锁乳突肌，它从二腹肌肩胛舌骨和后腹优越的​​肚子分开，并用拉钩容易暴露颈动脉的拉动。 </li><li>解剖颈动脉，将动脉购买结扎并固定在三个6-0丝缝合线。 </li></ol> 4.2）颈动脉的血流量测量<ol><li>将跨音速探头颈动脉周围，保持它的稳定，并使用超声波凝胶或盐水优化接触。 </li><li>颈动脉作为使用数据分析软件的跨音速设备的小屏幕上显示记录的血流速度</li><li>在完成测量后拆除探头</li></ol> 4.3）颈动脉压测量（CAP） <ol><li>结扎颈总动脉的远端和夹持其近端。 </li><li>将颈动脉周围2个固定缝合。用10-0的聚丙烯为入住的缝线。 </li><li>使在容器的前壁的小切口。 </li><li>将米勒导管，并修复它与预置缝线。 </li><li>记录在数据分析软件瓶盖。 </li></ol> 4.4）颈静脉血流测定<ol><li>解除颈静脉并放置跨音速气流探测器来测量流速。 </li><li>记录流率数据分析软件。 </li></ol> 4.5）中心静脉压测定（CVP） <ol><li>夹住颈静脉的近端和结扎的远端。 </li><li>切使用microscissors在容器的前壁的小切口。 </li><li>插入的填充流体的导管，并与预先放置缝合线固定。 </li><li>记录在数据分析软件的CVP。 </li></ol> 5，腹部手术的收购肝血流动力学的影响 5.1）船只识别<ol><li>使上腹部横切口。 </li><li> Eventerate肠内到左侧，并盖上湿纱布。 </li><li>辨认下腔静脉，门静脉，肝总动脉和肝固有动脉。 </li><li>整个监视过程中减少一些温生理盐水，在腹部和肠子每5分钟的表面上。 </li></ol>门脉血流5.2）测量<ol><li>解剖门静脉。 </li><li>放置在门静脉6-0丝线将所述流量探针时，以方便容器的升降。 </li><li>把跨音速气流探测器周围的门静脉，并测量其血流量。 </li><li>记录门静脉的血流量。 </li></ol>常见的肝动脉血流5.3）测量<ol><li>小心翼翼的解剖肝总动脉。 </li><li>放置在容器中围绕一个6-0丝线缝合，以方便容器的升降。 </li><li>放置在动脉周围的气流探测器。 </li><li>测量其血流量和获取数据。 </li></ol>门静脉压力5.4）测量（PVP） <ol><li>选择肠系膜静脉一些偏科，这水渠直接进入门静脉的一个分支。 </li><li>结扎所选肠系膜静脉的远端。确保结扎是接近肠道管。结扎的小分支</li><li>将2 Fixing使用静脉周围6-0普理灵缝线。这个过程的关键是避免结扎静脉时，触摸肠系膜动脉。 </li><li>夹住门静脉的近端。 </li><li>将用10-0的聚丙烯2逗留缝合。会出现一些出血，因为留缝线应穿过细肠系膜静脉的血管壁。 </li><li>就用microscissor倾斜成45度角的静脉小切口。 </li><li>将通过肠系膜静脉导管米拉尔进入门静脉和修复</li><li>记录门静脉压力。在该过程结束时，通过放血牺牲小鼠麻醉下。 </li></ol>

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作者宣称，他们没有竞争的经济利益。

肝脏血流动力学监测是一项重要的研究工具，在肝病和肝胆外科。收购肝血流动力学数据有助于表征循环系统肝胆程序的影响。还需要采集的肝血流动力学数据，研究了影响门脉压力和流量的门户， 如药物的疗效，需要在研究评估血管活性药物。 尽管小尺寸的重要参数，系统性和肝血流动力学可以在小鼠中进行监测。分别在协议中的关键步骤如下:首先，以放置在一个...

这部影片的总体目标是要证明一个实时监控程序，用于获取系统和肝脏血流动力学参数。开发这个程序的基本原理是在需要获得系统和肝脏血流动力学参数老鼠实验干预其巨大的价值。该过程可以应用于幼稚动物和期间或之后给定的肝胆实验外科干预，如肝部分切除，门静脉结扎和肝脏移植。 收购肝血流动力学数据，啮齿动物，需要提出的侵入性手术。肝脏灌注不能获得非侵入性。不过，也有收购全身血压的替代品。如尾袖套法8监测技术已被用于获取在大鼠和小鼠的血压。尾袖套法可以在有良知施加项动物。当测量血压时，动物需要被放置并固定在一个特定的不舒服的位置。在尾套设备的说明书，厂商指出，老鼠可能会变得紧张，并强调这可能削弱流通的尾巴。该种情况下，在尾巴获得的外周血压力可以比中央的血压低得多。 使用一系列传感器进行数据采集的集成多通道监测器进行全程监控程序。通过将导管插入后，在显微镜下仔细显微解剖和曝光的各容器中得到的血压。流速通过将跨音速流动探头每个容器周围测定。 我们已经报道了类似的术中监测程序导致了全面的一系列生理血流动力学数据堪比SINGL的老鼠来自其他团体7日报道E数据。因此，我们认为这个过程代表了良好的基础，使之适应鼠标，一个物种的10倍，比老鼠更小。在关键的区别与大鼠程序是利用Millar的导管，用于获取血压数据，而不是基于流体的导管系统。流动数据也获得了跨声速流动探头，只是很多小的比相应的大鼠血管。 由于动物的体积小，小鼠仪器在技术上是有挑战性的，但可行的。一旦仪器完成时，数据采集与主寿命数据分析是简单的，因为可以使用预定义的设置文件。设置文件，必须在一系列的实验中，开始一次定义，并且可以被存储并用于所有后续实验。 到现在为止，我们应用这个程序，以评估急性肝的实验血流动力学的影响。我们测得的CAP和PVP之前和紧接之后的70％部分肝切除（PH）和在夹紧/去夹紧的实验。我们夹在右叶表示质量接着位数的简短（5分钟）的夹紧和左外叶肝较完全的90％肝脏质量的20％的肝十二指肠韧带。去夹紧开始释放，从肝右叶后释放的中位数和左外叶夹。最大合模时间低于10分钟。

<table border="0" cellpadding="0" cellspacing="0" width="606">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:104px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PowerLab 16/30&#160;</td>
			<td style="width:104px;">ADInstruments</td>
			<td>PL3516</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Quad Bridge Amp</td>
			<td style="width:104px;">ADInstruments</td>
			<td>&#160;FE224</td>
			<td>Bridge amplifier&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Animal Bio Amp</td>
			<td style="width:104px;">ADInstruments</td>
			<td>&#160;FE136</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Needle Electrodes for FE136 (3 pk)</td>
			<td style="width:104px;">ADInstruments</td>
			<td>&#160;MLA1213</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Perivascular Flowmeter Module</td>
			<td style="width:104px;">Transonic</td>
			<td style="width:119px;">TS420&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Flowprobe MA0.5PSB/MA1PSB</td>
			<td style="width:104px;">Transonic</td>
			<td style="width:119px;">MA0.5PSB/MA1PSB</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">SPR-1000 Mouse Pressure Catheter</td>
			<td style="width:104px;">Millar instruments</td>
			<td>841-0001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">fluid filled catheter&#160;</td>
			<td>Terumo</td>
			<td style="width:119px;">SR+DU2619PX</td>
			<td>26G, 0.64&#215;19mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">micro scissors</td>
			<td>F&#183;S&#183;L</td>
			<td>No. 14058-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">micro serrefine</td>
			<td>F&#183;S&#183;L</td>
			<td>No.18055-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro clamps applicator</td>
			<td style="width:104px;">F&#183;S&#183;L</td>
			<td style="width:119px;">No. 18057-14</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Straight micro forceps</td>
			<td style="width:104px;">F&#183;S&#183;L</td>
			<td style="width:119px;">No. 00632-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Curved micro forceps</td>
			<td style="width:104px;">F&#183;S&#183;L</td>
			<td style="width:119px;">No. 00649-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">needle-holder</td>
			<td style="width:104px;">F&#183;S&#183;L</td>
			<td style="width:119px;">No. 12061-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:104px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">6-0 silk</td>
			<td style="width:104px;">ethicon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">6-0 prolene</td>
			<td style="width:104px;">ethicon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">7-0 prolene</td>
			<td style="width:104px;">ethicon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10-0 prolene</td>
			<td style="width:104px;">ethicon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tail cut-off device&#160;</td>
			<td style="width:104px;">Kent Scientific</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">www.kentscientific.com</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">LabChart7</td>
			<td style="width:104px;">ADInstruments</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">data&#160; analysis software&#160;</td>
		</tr>
	</tbody>
</table>

monitoring of systemic and hepatic hemodynamic parameters in mice

在实验研究中使用的小鼠模型是对肝脏的生理和病理障碍的研究有着重要意义。然而，由于鼠标的体积小，适合于鼠标术监测过程的技术细节很少说明。此前我们已经报道了监控程序，以获取血流动力学参数的影响。现在，我们适应的程序来获取小鼠全身及肝脏血流动力学参数，一个物种的10倍，比老鼠更小。该薄膜显示出动物的仪表以及评估在小鼠中的全身和肝血流动力学所需要的数据采集过程。重要的参数，包括体温，呼吸率和心脏速率，记录整个过程。全身血流动力学参数，包括颈动脉压（CAP）和中心静脉压（CVP）的。肝脏灌注参数包括门户vEIN压力（PVP），门脉流量以及肝总动脉（表1）的流量。仪器仪表和数据采集，记录的正常值是在1.5小时内完成。在此过程中的全身和肝血流动力学参数保持在正常范围内。 这个过程是具有挑战性的，但可行的。我们已经应用这个程序，以评估在正常小鼠以及在70％部分肝切除和肝瓣合模实验肝脏血流动力学。意思是PVP切除后（N = 20），为11.41±2.94 水柱这是显著高于对照组（P &lt;0.05）切除术（6.87±2.39 水柱 ）之前。肝叶夹紧实验的结果表明，该监视过程是敏感的，并适合于检测在入口压力和入口流速的微小变化。总之，这个过程是可靠的一个有经验的微型外科医生的手，但应限制为experiments地方是绝对需要的小鼠。

小鼠的重要参数，例如呼吸速率和心脏速率明显高于对大鼠高得多。平均全身血压和颈静脉压力类似于鼠值甚至类似于人的数据。 肝脏血流动力学的数据有明显的不同。我们从8只获得正常的值。在正常小鼠门脉血流之间的1.6〜2.3毫升/分钟之间。流入肝总动脉介于0.10至0.35毫升/分钟。在正常动物门静脉压力在大范围从4.4到11.2 水柱与8.09±2.47的平均值水柱 （表1...

Watch this Scientific Journal Video about Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice at JoVE.com

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

这部影片展示了如何获取小鼠全身及肝脏血流动力学。整个监视包括采集的重要参数，全身血压，中心静脉压，肝总动脉流量，和门静脉压，以及门户网站流量小鼠。

全身及肝脏血流动力学参数的监测小鼠

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

monitoring-of-systemic-and-hepatic-hemodynamic-parameters-in-mice

Research

JoVE Journal

Medicine

15.9K 次观看.  Jena University Hospital. 这部影片展示了如何获取小鼠全身及肝脏血流动力学。整个监视包括采集的重要参数，全身血压，中心静脉压，肝总动脉流量，和门静脉压，以及门户网站流量小鼠。 

视频: Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

One desirable endpoint of general anesthesia is the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is less straightforward than it is in human patients. A widely used behavioral surrogate for hypnosis in rodents is the loss of righting reflex (LORR), or the point at which the animal no longer responds to their innate instinct to avoid the vulnerability of dorsal recumbency. We have developed a system to assess LORR in 24 mice simultaneously while carefully controlling for potential confounds, including temperature fluctuations and varying gas flows. These chambers permit reliable assessment of anesthetic sensitivity as measured by latency to return of the righting reflex (RORR) following a fixed anesthetic exposure. Alternatively, using stepwise increases (or decreases) in anesthetic concentration, the chambers also enable determination of a population's sensitivity to induction (or emergence) as measured by EC50 and Hill slope. Finally, the controlled environmental chambers described here can be adapted for a variety of alternative uses, including inhaled delivery of other drugs, toxicology studies, and simultaneous real-time monitoring of vital signs.

Loss of the righting reflex has long served as a standard behavioral surrogate for unconsciousness, also called hypnosis, in laboratory animals. Alterations in volatile anesthetic sensitivity caused by pharmacological interventions can be detected with a carefully controlled high-throughput assessment system, which may be adapted for delivery of any inhaled therapeutic.

局部或全身药物干预后评估的变化小鼠中挥发性全身麻醉药敏感性

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

科研

医学

Clinical Assessment of Spatiotemporal Gait Parameters in Patients and Older Adults

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and neurological patients1-4, but also in healthy older adults5,6. The quantitative gait analysis described in this protocol is performed with a recently-introduced photoelectric system (see&#160;Materials table) which has the potential to be used in the clinic because it is portable, easy to set up (no subject preparation is required before a test), and does not require maintenance and sensor calibration. The photoelectric system consists of series of high-density floor-based photoelectric cells with light-emitting and light-receiving diodes that are placed parallel to each other to create a corridor, and are oriented perpendicular to the line of progression7. The system simply detects interruptions in light signal, for instance due to the presence of feet within the recording area. Temporal gait parameters and 1D spatial coordinates of consecutive steps are subsequently calculated to provide common gait parameters such as step length, single limb support and walking velocity8, whose validity against a criterion instrument has recently been demonstrated7,9. The measurement procedures are very straightforward; a single patient can be tested in less than 5 min&#160;and a comprehensive report can be generated in less than 1&#160;min.

This protocol is used to evaluate spatial and temporal gait variables of neurological/orthopedic patients and older persons by means of a recently-introduced floor-based photocell system.

时空步态参数的患者和老年人的临床评估

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Early Detection of Drug-Induced Renal Hemodynamic Dysfunction Using Sonographic Technology in Rats

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is estimated to contribute to 19- 25% of all clinical cases of acute kidney injury (AKI) in critically ill patients. AKI detection has historically relied on metrics such as serum creatinine (sCr) or blood urea nitrogen (BUN) which are demonstrably inadequate in full assessment of nephrotoxicity in the early phase of renal dysfunction. Currently, there is no robust diagnostic method to accurately detect hemodynamic alteration in the early phase of AKI while such alterations might actually precede the rise in serum biomarker levels. Such early detection can help clinicians make an accurate diagnosis and help in in decision making for therapeutic strategy. Rats were treated with Cisplatin to induce AKI. Nephrotoxicity was assessed for six days using high-frequency sonography, sCr measurement and upon histopathology of kidney. Hemodynamic evaluation using 2D and Color-Doppler images were used to serially study nephrotoxicity in rats, using the sonography. Our data showed successful drug-induced kidney injury in adult rats by histological examination. Color-Doppler based sonographic assessment of AKI indicated that resistive-index (RI) and pulsatile-index (PI) were increased in the treatment group; and peak-systolic velocity (mm/s), end-diastolic velocity (mm/s) and velocity-time integral (VTI, mm) were decreased in renal arteries in the same group. Importantly, these hemodynamic changes evaluated by sonography preceded the rise of sCr levels. Sonography-based indices such as RI or PI can thus be useful predictive markers of declining renal function in rodents. From our sonography-based observations in the kidneys of rats that underwent AKI, we showed that these noninvasive hemodynamic measurements may consider as an accurate, sensitive and robust method in detecting early stage kidney dysfunction. This study also underscores the importance of ethical issues associated with animal use in research.

Early stage hemodynamic dysfunction is critical to the development of kidney disease. Yet, detection methodologies are limited. Recent advances in sonography provide a noninvasive, accurate option for early detection of kidney injury. This study outlines a step-by-step, sonographic methodology for detecting kidney dysfunction using a drug-induced nephrotoxicity rat model.

大鼠使用超声技术药物引起的肾血流动力学障碍的早期检测

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is ...

The bm12 Inducible Model of Systemic Lupus Erythematosus (SLE) in C57BL/6 Mice

Systemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical and immunological manifestations. Several spontaneous and inducible animal models mirror common components of human disease, including the bm12 transfer model. Upon transfer of bm12 splenocytes or purified CD4 T cells, C57BL/6 mice rapidly develop large frequencies of T follicular helper cells (Tfh), germinal center (GC) B cells, and plasma cells followed by high levels of circulating anti-nuclear antibodies. Since this model utilizes mice on a pure C57BL/6 background, researchers can quickly and easily study disease progression in transgenic or knockout mouse strains in a relatively short period of time. Here we describe protocols for the induction of the model and the quantitation Tfh, GC B cells, and plasma cells by multi-color flow cytometry. Importantly, these protocols can also be used to characterize disease in most mouse models of SLE and identify Tfh, GC B cells, and plasma cells in other disease models.

The bm12 Inducible Model of Systemic Lupus Erythematosus SLE in C57BL/6 Mice

The transfer of bm12 lymphocytes into a C57BL/6 recipient is an established model of systemic lupus erythematosus. Here we describe how to initiate disease using this model and how to characterize T follicular helper cells, germinal center B cells and plasma cells by flow cytometry.

系统性红斑狼疮的BM12诱导模型（SLE）的C57BL / 6小鼠

Systemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical and immunological manifestations. Several spontaneous and inducible ...

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic study in the liver. Here, numerous assays are described to comprehensively measure the phenotype and parameters of hepatic steatosis in mouse and hepatocyte models. 
Combining the physiological, histological, and biochemical methods, this system can be used to assess the progress of hepatic steatosis. In vivo, the measurements of body weight and nuclear magnetic resonance (NMR) provide a general understanding of mice in a non-invasive manner. Hematoxylin and Eosin (H&amp;E) and Oil Red O staining determine the histological morphology and lipid deposition of liver tissue under nutrient overload conditions, such as high-fat diet feeding. Next, the total lipid contents are isolated by chloroform/methanol extraction, which are followed by a biochemical analysis for triglyceride and cholesterol. Moreover, mouse primary hepatocytes are treated with high glucose plus insulin to stimulate lipid accumulation, an efficient in vitro model to mimic diet-induced hyperglycemia and hyperinsulinemia in vivo. Then, the lipid deposition is measured by Oil Red O staining and chloroform/methanol extraction. Oil Red O staining determines intuitive hepatic steatotic phenotypes, while the lipid extraction analysis determines the parameters that can be analyzed statistically. The present protocols are of interest to scientists in the fields of fatty liver diseases, insulin resistance, and type 2 diabetes.

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Here, optimized methods to generate in vivo and in vitro models of hepatic steatosis and to analyze the steatotic phenotypes and related physiological parameters are described.

优化分析<em&gt;在体内</em&gt;和<em&gt;体外</em&gt;脂肪肝

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic ...

Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via a liver laceration that results in consistent blood loss, hemodynamic alterations, and survival.
Mice undergo a standardized resection of the left-middle lobe of the liver. They are allowed to bleed without mechanical intervention. Hemostatic agents can be administered as pre-treatment or rescue therapy depending on the interest of the investigator. During the time of hemorrhage, real-time hemodynamic monitoring via a left femoral arterial line is performed. Mice are then sacrificed, blood loss is quantified, blood is collected for further analysis, and organs are harvested for analysis of injury. Experimental design is described to allow for simultaneous testing of multiple animals.
Liver hemorrhage as a model of uncontrolled hemorrhage exists in the literature, primarily in rat and porcine models. Some of these models utilize hemodynamic monitoring or quantify blood loss but lack consistency. The present model incorporates quantification of blood loss, real-time hemodynamic monitoring in a murine model that offers the advantage of using transgenic lines and a high-throughput mechanism to further investigate the pathophysiologic mechanisms in uncontrolled hemorrhage.

Uncontrolled hemorrhage, an important cause of mortality among trauma patients, can be modeled using a standard liver laceration in a murine model. This model results in consistent blood loss, survival, and allows for testing hemostatic agents. This article provides the step-by-step process to perform this valuable model.

通过实时血液动力学监测在小鼠中通过肝脏浸润建立的不受控制的出血性休克

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via ...

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ICP waveform, amplitude of pulse (AMP), the correlation of ICP amplitude and ICP mean (RAP), pressure reactivity index (PRx), ICP and arterial blood pressure (ABP) wave amplitude correlation (IAAC), and so on, can reflect intracranial status, predict prognosis, and can also be used as guidance of proper treatment. However, most of the clinicians focus only on the mean ICP value while ignoring these parameters because of the limitations of the current devices. We have recently developed a multimodality monitoring system to address these drawbacks. This portable, user-friendly system will use a data collecting and storing device to continuously acquire patients&#39; physiological parameters first, i.e., ABP, ICP, and oxygen saturation, and then analyze these physiological parameters. We hope that the multimodality monitoring system will be accepted as a key measure to monitor physiological parameters, to analyze the current clinical status, and to predict the prognosis of the neurosurgical critical patients.

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

神经外科危重患者生理参数采集与分析的详细方案

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

小鼠颞叶癫痫的 Radiotelemetry 模型及脑电图监测

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

肝硬化大鼠门脉高压综合征的侵袭性血流动力学特征

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...