The authors have no acknowledgements.

所有研究均按照监管和制度指引完成。这项研究的动物方面是由肯特科技公司使用动物计划，由普渡大学动物护理和使用委员会（PACUC）认可的评估，并按照指南实验动物22的管理和使用进行。 注:本协议所使用的低流量数字麻醉系统配备有集成脉搏血氧饱和度。 1.建立集成式脉搏血氧仪低流量麻醉系统<ol><li>异氟醚交货<ol><li>选择载气源。要利用内部空气泵，旋进气口上背部，使内部泵吸入室内空气。 </li><li>连接活性炭罐到排气口。 </li><li> Y型适配器连接到低流量，数字麻醉系统的前面。使用颜色编码剪辑连接白色分支到t他鼻锥和蓝色分支到感应腔室。 </li><li>关闭白色夹子夹并打开蓝色夹子夹直接的气流室。 </li><li>选择2毫升注射器。 </li></ol></li><li>对于生理监测，使用集成式脉搏血氧仪<ol><li>连接脉搏血氧计爪子传感器端口上的低流量麻醉系统的背面，标有"AUX"。 </li></ol></li></ol> 2.配置设置<ol><li>麻醉<ol><li>电力麻醉系统，并进入设置菜单。按设置访问红色主菜单 &gt; 麻醉 &gt; 设置 。 </li><li>选择麻醉剂。按设置为突出型ANEST为红色。使用向上和向下箭头选择异氟醚。 </li><li>设置注射器大小。按设置以突出锡尔河大小 。使用向上和向下箭头选择2毫升注射器。 </li><li>设置空位。按设置为highlight红色设置空 。通过将注射器使注射器保持夹具上的注射器的金属套环就位固定在注射器保持块为空，完全压下玻璃注射器。按向上或向下箭头移动推块使推块使得与注射器活塞顶部的轻微接触。按选择设置空位。 </li><li>按设置以突出红色的删除 。取下注射器和填充用的瓶顶适配器异氟醚注射器。 </li><li>注射器连接到麻醉系统。 </li><li>总理麻醉递送管道。按设置以突出红色坯管 。向下按，直到麻醉穿过注射器进入蒸发器模块上的黑色配件。 </li><li>启用麻醉。按SET UP键突出显示启用红色。使用向上和向下箭头选择是 。按Run /返回，返回到主菜单。 </li><li>选择一个IR供应和每分钟通气量。按SET UP突出供气红色。使用向上和向下箭头键选择内置泵 。按设置以突出红色的分卷 。设定流速250毫升/分钟。 </li></ol></li><li>生理监控<ol><li>设定的最低检测心脏速率鼠标（240）。按SET UP键进入主菜单 &gt; MouseStat，用上下箭头设定最低心脏速率。 </li></ol></li></ol> 3.开始麻醉系统<ol><li>诱导鼠标<ol><li>按Run /返回两次进入运行模式，并开始气流。 </li><li>放置在感应室鼠标，关闭盖子紧。调节麻醉剂浓度旋钮3％。 </li><li>监测直到鼠标已经达到麻醉的所需平面，通过翻正反射损失决定。调节麻醉剂浓度是必要的。 </li><li>一旦动物失去其翻正反射一ND充分麻醉，打开麻醉浓度表盘为0％。笔者以前曾发现，让气流冲感应室30-60秒就足以清除室没有扭转麻醉深度1。 </li><li>快速打开白色夹子的空气直接到面罩，并关闭蓝色夹子导致感应室。 </li><li>从研究者打开密室走，移开鼠标，并立即适应鼻锥。 </li><li>围绕红外变暖垫，设置经由反馈回路直肠探头以维持体温在37℃的动物。 </li><li>当鼠标在鼻锥稳定，调整异氟烷的浓度至1.5％，或根据需要进行维护通过转动麻醉剂浓度旋钮。 </li><li>减少分钟通气量进行​​维护。的最小流量，以支持动物等于动物的分钟量1.5-2.2倍（在30 G鼠标，最小52毫升/分钟）。指制造商的说明为一个推荐流量具体设定为鼻锥样式，并根据需要进行调整。按SET UP键进入主菜单，然后按设定键直到分卷以红色突出显示。使用向上和向下箭头调整目标流量。按Run /返回，返回到主屏幕。 </li><li>确认麻醉深度为叉指捏过程中缺乏撤退反射的决定。应用眼药膏的眼睛，以防止在麻醉过程中干燥。 </li></ol></li></ol> 4.开始机能监测<ol><li>将传感器放到后掌垫。从而使红灯爪子底下，照亮了爪子传感器的位置。使用屏幕上的向上和向下箭头以显示oxiwave。动物现在已经安全使用低流量，注射器驱动，数字蒸发器麻醉。 </li></ol> 5.取下动物<ol><li>关闭麻醉系统。 <ol><li>要停止麻醉剂的递​​送，把麻醉剂浓度旋钮民（或0％）和口罩取下动物。 </li><li>监控麻醉恢复过程中的鼠标。一旦鼠标已经完全卧床，使其返回到笼子里。 </li></ol></li></ol>

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该项目由肯特科技公司，美国心脏协会CJ Goergen指出（SDG18220010）和普渡大学提供设备和资金的支持。作者克丽斯塔Bigiarelli和伊琳娜Toore是产生在这篇文章中使用的设备肯特科技公司的员工。这篇文章的开放获取出版由肯特科技公司赞助。

数字低流量麻醉系统允许用户有效地麻醉在非常低流速的小鼠，而无需使用任何压缩气体。这从标准无源汽化器，其中大部分需要在约500毫升/分钟的最小流速一个压缩气体源有很大不同。标准蒸发器利用缺乏层次之间的精确刻度盘，他们必须每年维修，以保持精度。注射器驱动麻醉剂系统可以在设定流量提供麻醉剂的特定浓度来计算所述注射器泵的确切需要的速度。常规标定是不必要的，从而导致额外的成本和节省时间。 建议的最低流量，以维持在非再呼吸电路的动物是1.5-2.2倍的动物的分钟量。在这项研究中使用了100毫升/分钟的流速超过了这个最小，以传递足够的麻醉的动物。流速设置搜索GS是这种麻醉剂递送技术关键的，作为流量直接相关的异氟烷用于给定的时间内的量。当在低流速时，这种技术可以大大降低使用过程中所需的异氟醚的量，而动物仍然有效1,19-21麻醉。 传统的蒸发器和低流量数字蒸发器之间的新设备的成本是相当的。然而，数字低流量麻醉系统具有提供任一异氟烷或七氟醚的能力。这消除了对指定异氟醚和七氟醚精度汽化器，减少了同时使用麻醉剂组初始设备成本的需求。蒸发器技术之间最近发表的比较都使用低流量数字蒸发器1,19,20时提出节约成本随着时间的推移。这些比较的结果可用于在一年的过程中潜在的成本节约近似。如舒米恩在2小时的增量，每周5天的52周进行典型使用设置，传统的异氟烷蒸发器将消耗3.8升异氟烷，或12个250 ml瓶装。在相同的频率中使用的低流量数字汽化器将消耗只是0.32 L或二份各250ml的瓶子。活性碳罐的消耗也减少了。假设每个罐持有50克清理废气，传统的蒸发器，将填补约21木炭罐超过一年的课程。相比之下，低流量数字汽化器将需要6个或更少。传统的蒸发器将需要每年约5大气瓶，每9,500 L的内置气泵，在数字低流量汽化器某些型号的容量，省去了压缩气体的要求。如果压缩气体都被使用，则系统将使用每年1只1气缸。 该技术可以根据需要进行修改。低流量数字vaporiz器允许用户快速和精确地调整麻醉深度。如果麻醉深度必须增加或减小时，用户​​可以使用拨号的系统的顶部增加0.1％的增量麻醉浓度。根据需要在整个过程也可调节的流速。该协议采用2毫升注射器，但更大的注射器规格可供较长的过程。内部空气泵为用户提供了麻醉动物，而不需要一个压缩气体源的选项。对于需要压缩气体或补充氧气程序时，用户具有对气体源到低流量系统连接，而不是使用周围空气的选项。用户可以继续对整个过程提供所选择的空气源，或可以在内部泵并根据需要在压缩气体源之间切换。例如，用户可将系统设置为诱导和维持期间通过内部泵输送室内空气，但提供补充氧恢复期间根。 虽然有很多优点，使用低流量数字蒸发器，也有局限性。因为冲洗阀不包括，开口之前手动冲洗用清洁空气腔室是吹扫感应腔室的唯一途径。这个系统被设计成在仅低流速操作和不提供上述800毫升/分钟，而传统的蒸发器可与流率被用于高达10升/分钟流速麻醉。因此，这种特定的系统是仅适用于小动物物种。另外，相对于传统的蒸发器系统中保持较少的麻醉剂。可能存在的注射器必须在手术期间再填充的情况。然而，可以通过填写预第二注射​​器附近，以取代空注射器可​​以减少再填充期间的延迟。注射器尺寸可达10 mL可用来减少需要重新填充注射器中端程序。最后，与传统的蒸发器中，低FL流数字蒸发器需要的电力。电池是可用于实例使用，其中电力不可用或在停电的情况下。 以前的研究已经表明，低流量数字系统相比，传统的麻醉系统1,19,20消耗更少的异氟烷，载气，和木炭罐。清除的麻醉气体的减少也能识别垃圾麻醉气体的减少，虽然需要在这些领域进一步开展工作。红外气体光谱可用于监测废物异氟醚的生产，和剂量计徽章可以用于量化在将来的比较异氟烷暴露于实验室人员。 总之，该技术对于麻醉剂递送将是执行啮齿动物麻醉由于改善的安全性，有效性和精度比传统系统组是有益的。

有迹象表明，0.5-10升/分钟2流量之间的操作可兽用许多精密喷雾器。这些流量是不理想的老鼠相比，它们的小每分钟呼吸量范围为高。高流速不会在兽医实践建议由于它们促进低温和呼吸道3,4-干燥的。此外，许多普通兽医汽化器制造商的手册警告高流速可能导致背压的波动的发生增加。还已经表明，许多标准汽化器变得不准确低于500ml /分的流速，和该速率被认为是在兽医领域5-7最小流率。 动物可以保持在T形件的电路，或使用低至1.5-2.2倍动物的分钟量8-10流速改性贝恩电路。这些流量被认为是sufficien吨至到期防止气体再呼吸，防止血液中二氧化碳浓度8的增加。使用该流速建议，30 G鼠标可维持在流速低高达52毫升/分钟，高于最低传统蒸发器的接受500ml /分少了近十倍。 而传统的汽化器取决于气流并用于被动麻醉剂汽化大气压，直喷汽化器测量总新鲜气体流并直接注入蒸汽到气体流2。一些直接喷射喷雾器利用注射器泵施用麻醉剂到气体流中。计算机化控制允许这些系统来自动调节注射泵的速度来注入达到麻醉剂的所需浓度所需的液体试剂的量。注射器驱动的蒸发器可用，并批准用于临床和儿童使用，许多类似的配置视为美学在临床实践中11-16节约型的设备。他们的批准后不久，随着注射泵喷雾器麻醉节约设备被改编为在动物研究8,17,18使用。不同于传统的蒸发器，利用一个注射泵直接喷射系统不是由最小流率，以保持精度的限制。出于这个原因，这种技术非常适用于啮齿动物麻醉并在低流率是必要的其它实例中使用。与此蒸发器设计相关的益处和潜在的成本节约启发专门为啮齿类1,19,20设计新麻醉系统的发展。这种新的系统还包括一个内置的气泵，允许用户管理麻醉而不需要压缩气体源。作为一个额外的好处，该系统是预先校准为与两个异氟醚和七氟醚的使用。凭借在实验动物领域引进这项技术的蒸发器，我t是现在有可能在流速接近推荐的水平麻醉小实验室动物，而不需要压缩气体。

<table border="0" cellpadding="0" cellspacing="0" width="873">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:283px;">Anesthetic Equipment</td>
			<td style="width:131px;"></td>
			<td style="width:119px;"></td>
			<td style="width:340px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">SomnoSuite Low-Flow Digital Anesthesia System</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">SOMNO</td>
			<td style="width:340px;">Includes anti-spill, anti-vapor bottle top adapter; Y adapter tubing; charcoal scavenging filter</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">MouseSTAT Pulse Oximeter &#38; Heart Rate Monitor</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">SS-MSTAT-Module</td>
			<td style="width:340px;">Integrated into SomnoSuite</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">MouseSTAT Mouse Paw Sensor</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">MSTAT-MSE</td>
			<td style="width:340px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">2mL Glass Syringe</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">SOMNO-2ML</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">Low-Cost Induction Chamber, 0.5L</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">SOMNO-0705</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">Low Profile Facemask, x-small</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">SOMNO-0304</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:283px;">Animal Warming</td>
			<td style="width:131px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:283px;">PhysioSuite Physiological Monitoring System with RightTemp Homeothermic Warming</td>
			<td style="width:131px;">Kent Scientific Corporation</td>
			<td style="width:119px;">PS-RT</td>
			<td style="width:340px;">Includes infrared warming pad, rectal probe, and pad temperature probe</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">Anesthetic Agents 
			and Medications</td>
			<td style="width:131px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:283px;">Isoflurane (250mL bottle)</td>
			<td style="width:131px;">Piramal Healthcare</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:283px;">Puralube Opthalmic Ointment</td>
			<td style="width:131px;">Perrigo</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

use of a low-flow digital anesthesia system for mice and rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

动物 3成年C57 / BL6NTac雌性小鼠（Taconic，年龄6-7周; 15重量+/- 1克）麻醉，并以1.3-1.5％异氟烷维持而心脏速率，氧饱和度，和呼吸速率进行了监测。所有小鼠的鼠无病原体的常规供应商测试抵达设施之前所确定。这些动物群养在了显微笼养并提供了一​​瓶免费获取标准啮齿动物食物和水。 使用异氟醚低流量麻醉系统测量在使用期间残留在注射器麻醉剂的量。注射器中的体积，如通过麻醉系统测量，指出作为动物在维持期结束时转移到鼻锥，并再次。最终体积W从初始体积减去量化维修期间（ 图1）所消耗的麻醉剂的量。 生理参数心脏率， 血氧饱和度和呼吸速率通过脉搏血氧饱和度（ - 4 图2）维护期间进行了监测。体温是通过一个红外暖垫保持在37.5℃。每只小鼠成功地保持以100ml室内空气的/分低流速麻醉的手术平面下60分钟后，通过从一个交叉指捏缺乏撤退反射的测定。鼠标没有清醒或在维修期间间歇应用趾间捏回应。动物的心脏速率（ 图2），血氧（ 图3），和呼吸率（ 图4）保持RELAT整个研究ively稳定。由于动物和传感器的定位，从鼠标1和鼠标3的呼吸速率的信号是间歇性的，并测量被中断。当动物的定位调整，信号改善，所测量的呼吸率是可比的他人在类似的时间点。低流量数字麻醉系统维护期间（ 图1）的60分钟期间使用的0.63毫升异氟醚的平均值。 <img alt="图1" src="/files/ftp_upload/54436/54436fig1.jpg" /> 图 1: 异氟醚异氟醚的使用在毫升超过使用数字低流量麻醉系统麻醉维持1小时用于三种不同的小鼠量<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54436/54436fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图 2: 心率三只老鼠在每分钟（BPM）的初始麻醉诱导与数字低流量麻醉系统后5-60分钟节拍心脏速率<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54436/54436fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54436/54436fig3.jpg" /> 图3:。 血氧饱和度三只老鼠初始麻醉诱导与数字低流量麻醉系统后5-60分钟的血氧饱和度（％） <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54436/54436fig3large.jpg" target="_blank">点击此处查看该图的放大版本。</a> 图4:。呼吸率三只老鼠在每分钟（BPM）的初始麻醉诱导与数字低流量麻醉系统后5-60分钟呼吸的呼吸速率<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54436/54436fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Use of a Low-flow Digital Anesthesia System for Mice and Rats at JoVE.com

Use of a Low-flow Digital Anesthesia System for Mice and Rats

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

低流量数字麻醉系统的小鼠和大鼠的用途

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

use-of-a-low-flow-digital-anesthesia-system-for-mice-and-rats

Nanyang Technological University

Research

JoVE Journal

Medicine

17.8K 次观看.  Purdue University. Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer. 

视频: Use of a Low-flow Digital Anesthesia System for Mice and Rats

A Novel Rescue Technique for Difficult Intubation and Difficult Ventilation

We describe a novel non surgical technique to maintain oxygenation and ventilation in a case of difficult intubation and difficult ventilation, which works especially well with poor mask fit.
Can not intubate, can not ventilate" (CICV) is a potentially life threatening situation. In this video we present a simulation of the technique we used in a case of CICV where oxygenation and ventilation were maintained by inserting an endotracheal tube (ETT) nasally down to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.
A 13 year old patient was taken to the operating room for incision and drainage of a neck abcess and direct laryngobronchoscopy. After preoxygenation, anesthesia was induced intravenously. Mask ventilation was found to be extremely difficult because of the swelling of the soft tissue. The face mask could not fit properly on the face due to significant facial swelling as well. A direct laryngoscopy was attempted with no visualization of the larynx. Oxygen saturation was difficult to maintain, with saturations falling to 80%. In order to oxygenate and ventilate the patient, an endotracheal tube was then inserted nasally after nasal spray with nasal decongestant and lubricant. The tube was pushed gently and blindly into the hypopharynx. The mouth and nose of the patient were sealed by hand and positive pressure ventilation was possible with 100% O2 with good oxygen saturation during that period of time. Once the patient was stable and well sedated, a rigid bronchoscope was introduced by the otolaryngologist showing extensive subglottic and epiglottic edema, and a mass effect from the abscess, contributing to the airway compromise. The airway was secured with an ETT tube by the otolaryngologist.This video will show a simulation of the technique on a patient undergoing general anesthesia for dental restorations.

We describe a technique to maintain oxygenation and ventilation using an endotracheal tube inserted nasally to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.

一个新型插管困难和困难通风救援技术

We describe a novel non surgical technique to maintain oxygenation and ventilation in a case of difficult intubation and difficult ventilation, which ...

科研

医学

Guidelines for Elective Pediatric Fiberoptic Intubation

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small babies will desaturate very quickly if ventilation is interrupted mainly to high metabolic rate. We describe guidelines to perform a safe fiberoptic intubation while maintaining spontaneous breathing throughout the procedure. Steps requiring the use of propofol pump, fentanyl, glycopyrrolate, red rubber catheter, metal insuflation hook, afrin, lubricant and lidocaine spray are shown.

We describe guidelines to perform a safe and efficient elective fiberoptic intubation in pediatric patients while maintaining spontaneous ventilation.

选修儿科光纤插管指南

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small ...

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. Cardioprotection by ischemic preconditioning (IP) remains an area of intense investigation. To further elucidate its molecular basis, the use of knockout mouse studies is particularly important 7, 14, 30, 39. Despite the fact that previous studies have already successfully performed cardiac ischemia and reperfusion in mice, this model is technically very challenging. Particularly, visual identification of the coronary artery, placement of the suture around the vessel and coronary occlusion by tying off the vessel with a supported knot is technically difficult. In addition, re-opening the knot for intermittent reperfusion of the coronary artery during IP without causing surgical trauma adds additional challenge. Moreover, if the knot is not tied down strong enough, inadvertent reperfusion due to imperfect occlusion of the coronary may affect the results. In fact, this can easily occur due to the movement of the beating heart.
Based on potential problems associated with using a knotted coronary occlusion system, we adopted a previously published model of chronic cardiomyopathy based on a hanging weight system for intermittent coronary artery occlusion during IP 39. In fact, coronary artery occlusion can thus be achieved without having to occlude the coronary by a knot. Moreover, reperfusion of the vessel can be easily achieved by supporting the hanging weights which are in a remote localization from cardiac tissues.
We tested this system systematically, including variation of ischemia and reperfusion times, preconditioning regiments, body temperature and genetic backgrounds39. In addition to infarct staining, we tested cardiac troponin I (cTnI)
as a marker of myocardial infarction in this model. In fact, plasma levels of cTnI correlated with infarct sizes (R2=0.8). Finally, we could show in several studies that this technique yields highly reproducible infarct sizes during murine IP and myocardial infarction6, 8, 30, 40, 41. Therefore, this technique may be helpful for researchers who pursue molecular mechanisms involved in cardioprotection by IP using a genetic approach in mice with targeted gene deletion. Further studies on cardiac IP using transgenic mice may consider this technique.

A murine model for myocardial ischemia and ischemic preconditioning is an important tool study cardioprotective mechanisms in vivo. Here, we report an easy applicable in situ model for cardiac IP using a hanging-weight system for coronary artery occlusion.

悬重系统的使用在小鼠冠状动脉闭塞

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. ...

Use of a Hanging-weight System for Isolated Renal Artery Occlusion

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients revealed that only a mild increase in serum creatinine levels (0.3 to 0.4 mg/dl) is associated with a 70% greater risk of death than in persons without any increase 1. Along these lines, surgical procedures requiring cross-clamping of the aorta and renal vessels are associated with a renal failure rates of up to 30% 4. Similarly, AKI after cardiac surgery occurs in over 10% of patients under normal circumstances and is associated with dramatic increases in mortality. AKI are also common complications after liver transplantation. At least 8-17% of patients end up requiring renal replacement therapy 5. Moreover, delayed graft function due to tubule cell injury during kidney transplantation is frequently related to ischemia-associated AKI 6. Moreover, AKI occurs in approximately 20% of patients suffering from sepsis 6.The occurrence of AKI is associated with dramatic increases of morbidity and mortality 1. Therapeutic approaches are very limited and the majority of interventional trials in AKI have failed in humans. Therefore, additional therapeutic modalities to prevent renal injury from ischemia are urgently needed 3, 7-9. 
To elucidate mechanisms of renal injury due to ischemia and possible therapeutic strategies murine models are intensively required 7-13. Mouse models provide the possibility of utilizing different genetic models including gene-targeted mice and tissue specific gene-targeted mice (cre-flox system). However, murine renal ischemia is technically challenging and experimental details significantly influence results. We performed a systematic evaluation of a novel model for isolated renal artery occlusion in mice, which specifically avoids the use of clamping or suturing the renal pedicle 14. This model requires a nephrectomy of the right kidney since ischemia can be only performed in one kidney due to the experimental setting. In fact, by using a hanging-weight system, the renal artery is only instrumented once throughout the surgical procedure. In addition, no venous or urethral obstruction occurs with this technique. We could demonstrate time-dose-dependent and highly reproducible renal injury with ischemia by measuring serum creatinine. Moreover, when comparing this new model with conventional clamping of the whole pedicle, renal protection by ischemic preconditioning is more profound and more reliable. Therefore his new technique might be useful for other researchers who are working in the field of acute kidney injury.

A precise murine model for acute kidney injury (AKI) due to ischemia is an important tool to investigate acute kidney injury and possibly find therapeutic tools to treat renal injury. The hanging weight system offers a tool for immediate and reliable renal artery occlusion and reperfusion without causing renal congestion.

一个悬重系统使用隔离肾动脉闭塞

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients ...

Use of a Hanging-weight System for Liver Ischemia in Mice

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and can result in liver failure which has a high mortality rate1-2. Therefore murine studies of hepatic ischemia have become an important field of research by providing the opportunity to utilize pharmacological and genetic studies3-9. Specifically, conditional mice with tissue specific deletion of a gene (cre, flox system) provide insights into the role of proteins in particular tissues10-13 . Because of the technical difficulty associated with manually clamping the portal triad in mice, we performed a systematic evaluation using a hanging-weight system for portal triad occlusion which has been previously described3. By using a hanging-weight system we place a suture around the left branch of the portal triad without causing any damage to the hepatic lobes, since also the finest clamps available can cause hepatic tissue damage because of the close location of liver tissue to the vessels. Furthermore, the right branch of the hepatic triad is still perfused thus no intestinal congestion occurs with this technique as blood flow to the right hepatic lobes is preserved. Furthermore, the portal triad is only manipulated once throughout the entire surgical procedure. As a result, procedures like pre-conditioning, with short times of ischemia and reperfusion, can be easily performed. Systematic evaluation of this model by performing different ischemia and reperfusion times revealed a close correlation of hepatic ischemia time with liver damage as measured by alanine (ALT) and aspartate (AST) aminotransferase serum levels3,9. Taken together, these studies confirm highly reproducible liver injury when using the hanging-weight system for hepatic ischemia and intermittent reperfusion. Thus, this technique might be useful for other investigators interested in liver ischemia studies in mice. Therefore the video clip provides a detailed step-by-step description of this technique.

We established a novel murine model of a hanging weight system for portal triad occlusion. This technique may be useful for future investigations of ischemia in murine hepatic models.

使用吊重系统在小鼠肝缺血

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

脓毒症动物模型，用于评估新型草药疗法

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Intraoperative Detection of Subtle Endometriosis: A Novel Paradigm for Detection and Treatment of Pelvic Pain Associated with the Loss of Peritoneal Integrity

Endometriosis is a common disease affecting 40 to 70% of reproductive-aged women with chronic pelvic pain (CPP) and/or infertility. The purpose of this study was to demonstrate the use of a blue dye (methylene blue) to stain peritoneal surfaces during laparoscopy (L/S) to detect the loss of peritoneal integrity in patients with pelvic pain and suspected endometriosis. Forty women with CPP and 5 women without pain were evaluated in this pilot study. During L/S, concentrated dye was sprayed onto peritoneal surfaces, then aspirated and rinsed with Lactated Ringers solution. Areas of localized dye uptake were evaluated for the presence of visible endometriotic lesions. Areas of intense peritoneal staining were resected and some fixed in 2.5% buffered gluteraldehyde and examined by scanning (SEM) electron microscopy. Blue dye uptake was more common in women with endometriosis and chronic pelvic pain than controls (85% vs. 40%). Resection of the blue stained areas revealed endometriosis by SEM and loss of peritoneal cell-cell contact compared to normal, non-staining peritoneum. Affected peritoneum was associated with visible endometriotic implants in most but not all patients. Subjective pain relief was reported in 80% of subjects. Based on scanning electron microscopy, we conclude that endometrial cells extend well beyond visible implants of endometriosis and appear to disrupt the underlying mesothelium. Subtle lesions of endometriosis could therefore cause pelvic pain by disruption of peritoneal integrity, allowing menstrual or ovulatory blood and associated pain factors access to underlying sensory nerves. Complete resection of affected peritoneum may provide a better long-term treatment for endometriosis and CPP. This simple technique appears to improve detection of subtle or near invisible endometriosis in women with CPP and minimal visual findings at L/S and may serve to elevate diagnostic accuracy for endometriosis at laparoscopy.

Loss of peritoneal integrity provides a new paradigm to understand and treat chronic pelvic pain in women with mild forms of endometriosis and can be easily detected using intraoperative instillation of dye at the time of laparoscopy.

术中的微妙的子宫内膜异位症的检测:A新型范式为相关联的与的的腹膜的完整性之亏损盆腔疼痛的的的的检测和治疗的

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known risk factor for glaucoma. This protocol demonstrates a step-by-step guide on this technique in Long Evans rats and C57BL/6 mice. Under general anesthesia, a "purse-string" suture is applied on the conjunctiva, around the equator and behind the limbus of the eye. The fellow eye serves as an untreated control. Over the duration of our study, which was a period of 8 weeks for rats and 12 weeks for mice, IOP remained elevated, as measured regularly by rebound tonometry in conscious animals without topical anesthesia. In both species, the sutured eyes showed electroretinogram features consistent with preferential inner retinal dysfunction. Optical coherence tomography showed selective thinning of the retinal nerve fiber layer. Histology of the rat retina in cross-section found reduced cell density in the ganglion cell layer, but no change in other cellular layers. Staining of flat-mounted mouse retinae with a ganglion cell specific marker (RBPMS) confirmed ganglion cell loss. The circumlimbal suture is a simple, minimally invasive and cost-effective way to induce ocular hypertension that leads to ganglion cell injury in both rats and mice.

Chronic ocular hypertension is induced by applying a circumlimbal suture in rats and mice, leading to functional and structural deterioration of the retinal ganglion cells consistent with glaucoma.

Circumlimbal 缝合法诱导大鼠和小鼠青光眼模型的建立

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

正交心脏移植的临床前猪模型

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare recommendations, animals are warmed and monitored during procedures requiring anesthesia. Common anesthesia and physiological monitoring equipment include gas tanks, anesthetic vaporizers and stands, warming controllers and pads, mechanical ventilators, and pulse oximeters. A computer is also necessary for data collection and to run equipment software. In smaller spaces or when performing field work, it can be challenging to configure all this equipment in limited space.
The goal of this protocol is to demonstrate best practices for use of a low-flow digital vaporizer using both compressed oxygen and room air, along with an integrated mechanical ventilator, pulse oximeter, and far infrared warming as an all-inclusive anesthesia and physiological monitoring suite ideal for rodents.

Here, we present a protocol to safely and effectively administer anesthetic gas to mice using a digital, low flow anesthesia system with integrated ventilator and physiological monitoring modules.

使用啮齿动物综合低流量麻醉蒸发器、呼吸机和生理监测系统

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare ...