作者没有承认。

所有动物研究都得到了普渡动物护理和使用委员会的批准。
1. 低流量蒸发器的设置
<ol><li>伊索夫卢兰或塞沃夫鲁兰交付<ol>
<li>选择载体气源。要利用内部空气泵，请从系统背面的入口端口取出红色盖，使系统能够接收室内空气。要使用压缩气体，请使用设置为 15 PSI 的压力调节器或减压器，并连接到系统背面的压缩气体端口。</li>
		<li>将木炭罐连接到排气口。</li>
		<li>将附件连接器连接到系统前面的激励和过期端口。将感应室连接到带有蓝色夹子的树枝和鼻锥连接到带有白色夹子的树枝（图 1）。</li>
	</ol>
</li>
	<li>用于机械通风<ol>
<li>将连接连接器管连接到黄色编码剪辑（图 2）。</li>
		<li>通过执行死空间校准来校准呼吸机。从 通风运行屏幕，触摸 设置，然后 校准和测试。选择 死空间校准 并按 拨号 B。</li>
	</ol>
</li>
	<li>用于脉冲氧化<ol>
<li>将传感器连接到系统背面的端口，标记为 MouseSTAT。</li>
	</ol>
</li>
	<li>用于变暖<ol>
<li>将加热垫连接到系统前面的"垫电源"端口。</li>
		<li>将一个传感器连接到"身体传感器"端口，另一个传感器连接到"垫传感器"端口。将垫传感器固定到加热垫上。</li>
	</ol>
</li>
</ol>2. 配置设置
<ol><li>麻醉<ol>
<li>麻醉系统电源。从 阿内斯特运行屏幕，触摸 设置。</li>
		<li>选择麻醉剂。触摸 类型阿内斯特，然后转动拨号B选择 伊索夫卢兰 或 塞沃夫鲁兰。</li>
		<li>设置注射器大小。触摸 注射器大小，然后转动拨号B以选择大小。</li>
		<li>触摸返回返回到 阿内斯特运行屏幕。</li>
		<li>使用瓶顶适配器，用麻醉剂填充注射器。</li>
		<li>将注射器连接到麻醉系统。如果需要，触摸 删除 以向后移动推手块。</li>
		<li>将注射器主热。触摸并保持 Prime 向前移动推手块，直到推手块接触注射器柱塞的顶部。按住 Prime 按钮时转动 B 会调节推手块速度。</li>
	</ol>
</li>
	<li>用于机械通风<ol>
<li>触摸 文特运行屏幕 选项卡，然后 设置。</li>
		<li>触摸 体重 并输入动物的重量。</li>
		<li>触摸 优先选择 音量或压力控制通风。体重设置自动设置适当的呼吸速率和潮汐体积。</li>
	</ol>
</li>
	<li>用于脉冲氧化<ol>
<li>触摸 Oxi 运行屏幕 选项卡，然后 设置。</li>
		<li>触摸 HR 并转动拨号 B 以设置允许的最低心率读数。预设可用。</li>
	</ol>
</li>
	<li>用于变暖<ol>
<li>从 暖运行屏幕，触摸 设置。选择加热方法和目标温度设置。</li>
	</ol>
</li>
</ol>3. 开始麻醉分娩
<ol><li>麻醉鼠标<ol>
<li>从 Anest 运行屏幕，触摸 启动感应 开始气流。默认感应流速为 500 mL/min。转动拨号 A 可根据需要调整流速。</li>
		<li>将鼠标放在感应室中，紧紧地合上盖子。将 麻醉剂浓度 表盘调整为 3% 的异氟化剂。</li>
		<li>监测，直到小鼠到达所需的麻醉平面，由呼吸速率下降和右反射丧失时，室倾斜。根据需要调整 麻醉剂浓度 表盘。</li>
		<li>一旦动物失去了右反射，并充分麻醉，触摸 停止感应。</li>
		<li>如果需要，触摸 冲洗室 以清空残留麻醉气体的腔室。</li>
		<li>打开通往鼻锥的夹子，并关闭通向腔室的夹子。</li>
		<li>触摸 启动鼻锥。体重设置确定鼻锥流速率，但可以通过转动拨号 A 手动调整。</li>
		<li>立即安装鼻锥，并将动物放在红外加热垫上。</li>
		<li>将动物传感器插入作为直肠探头。</li>
	</ol>
</li>
</ol>4. 开始机械通风
<ol><li>灌管动物。<ol>
<li>将动物转移到插管阶段，同时保持动物麻醉。</li>
		<li>使用固定在垂直插管阶段的螺纹（图3）将动物从上切口吊起。</li>
		<li>轻轻地将动物的舌头放在一边，使用灌装套件中提供的灯光可视化气管。</li>
		<li>小心地插入气管，通过将小气囊连接到管子并检查肺部是否膨胀来验证正确位置。</li>
	</ol>
</li>
	<li>将内脏管连接到通风管。</li>
	<li>触摸 停止鼻锥，然后触摸 启动呼吸机。 注意:体重设置自动确定适当的呼吸速率和潮汐量。要执行压力控制通风，将目标吸入压力设置在 15-18 厘米 H2O 之间。根据手术方案需要对呼吸机设置进行调整。</li>
</ol>5. 开始生理监测
<ol><li>将传感器放在动物的后爪上（图4）。脉冲氧化计将自动开始读取HR和SPO2。 触摸 Oxi 运行屏幕选项卡 以查看脉冲氧化数据。</li>
</ol>

<ol>
	<li>El-Attar, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guided+isoflurane+injection+in+a+totally+closed+circuit.">Guided isoflurane injection in a totally closed circuit.</a> Anaesthesia. 46 (12), 1059-1063 (1991).</li><li>Lockwood, G., Chakrabarti, M. K., Whitwam, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+computer-controller+closed+anaesthetic+breathing+system.">A computer-controller closed anaesthetic breathing system.</a> Anaesthesia. 48 (8), 690-693 (1993).</li><li>Lowe, H. J., Cupic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-regulated+automated+anesthesia+(Abstract).">Dose-regulated automated anesthesia (Abstract).</a> British Journal of Clinical Pharmacologyl. 12 (2), 281-282 (1971).</li><li>Walker, T. J., Chackrabarti, M. K., Lockwood, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+of+desflurane+during+anaesthesia.">Uptake of desflurane during anaesthesia.</a> Anaesthesia. 51 (1), 33-36 (1996).</li><li>Weingarten, M., Lowe, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+circuit+injection+technic+for+syringe-measured+administration+of+methoxyflurane:+a+new+dimension+in+anesthesia.">A new circuit injection technic for syringe-measured administration of methoxyflurane: a new dimension in anesthesia.</a> Anesthesia &amp; Analgesia. 52 (4), 634-642 (1973).</li><li>Enlund, M., Wiklund, L., Lambert, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+device+to+reduce+the+consumption+of+a+halogenated+anaesthetic+agent.">A new device to reduce the consumption of a halogenated anaesthetic agent.</a> Anaesthesia. 56 (5), 429-432 (2001).</li><li>Kelly, J. M., Kong, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+ten+isoflurane+vaporisers+in+current+clinical+use.">Accuracy of ten isoflurane vaporisers in current clinical use.</a> Anaesthesia. 66 (8), 682-688 (2011).</li><li>Matsuda, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NARCOBIT+-+A+newly+developed+inhalational+anesthesia+system+for+mice.">NARCOBIT - A newly developed inhalational anesthesia system for mice.</a> Experimental Animals. 56 (2), 131-137 (2007).</li><li>Soro, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+accuracy+of+the+anesthetic+conserving+device+(Anaconda)+as+an+alternative+to+the+classical+vaporizer+in+anesthesia.">The accuracy of the anesthetic conserving device (Anaconda) as an alternative to the classical vaporizer in anesthesia.</a> Anesthesia &amp; Analgesia. 111 (5), 1176-1179 (2010).</li><li>Ward, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Physical principles and maintenance. Anaesthetic equipment. , (1985).</li><li>Ambrisko, T. D., Klide, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+isoflurane+and+Sevoflurane+vaporizers+over+a+wide+range+of+oxygen+flow+rates.">Evaluation of isoflurane and Sevoflurane vaporizers over a wide range of oxygen flow rates.</a> American Journal of Veterinary Research. 67 (6), 936-940 (2006).</li><li>Damen, F. W., Adelsperger, A. R., Wilson, K. E., Goergen, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+traditional+and+integrated+digital+anesthetic+vaporizers.">Comparison of traditional and integrated digital anesthetic vaporizers.</a> Journal of the American Association for Laboratory Animal Science. 54 (6), 756-762 (2015).</li><li>Adelsperger, A. R., Bigiarelli-Nogas, K. J., Toore, I., Goergen, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+Low-flow+Digital+Anesthesia+System+for+Mice+and+Rats.">Use of a Low-flow Digital Anesthesia System for Mice and Rats.</a> Journal of Visualized Experiments. (115), e54436 (2016).</li><li>Flecknell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Laboratory animal anaesthesia. , (2009).</li><li>Mapleson, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elimination+of+rebreathing+in+various+semiclosed+anaesthetic+systems.">The elimination of rebreathing in various semiclosed anaesthetic systems.</a> British Journal of Anaesthesia. 26 (5), 323-332 (1954).</li><li>Chakravarti, S., Basu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modern+Anaesthesia+Vapourisers.">Modern Anaesthesia Vapourisers.</a> Indian Journal of Anaesthesia. 57 (5), 464-471 (2013).</li><li>Mullin, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+anesthesia+carrier+gas+on+in+vivo+circulation+times+of+ultrasound+microbubble+contrast+agents+in+rats.">Effect of anesthesia carrier gas on in vivo circulation times of ultrasound microbubble contrast agents in rats.</a> Contrast Media &amp; Molecular Imaging. 6 (3), 126-131 (2011).</li><li>Flores, J. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+anesthetic+agent+and+carrier+gas+on+blood+glucose+and+tissue+uptake+in+mice+undergoing+dynamic+FDG-PET+imaging:+sevoflurane+and+isoflurane+compared+in+air+and+in+oxygen.">The effects of anesthetic agent and carrier gas on blood glucose and tissue uptake in mice undergoing dynamic FDG-PET imaging: sevoflurane and isoflurane compared in air and in oxygen.</a> Molecular Imaging and Biology. 10 (4), 192 (2008).</li><li>Carroll, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Small Animal Anesthesia and Analgesia. , (2008).</li><li>Thomas, J., Lerche, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Anesthesia and Analgesia for Veterinary Technicians, 4th ed. 335, (2011).</li><li>McKelvey, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Veterinary Anesthesia and Analgesia. , (2003).</li><li>Tranquilli, W. J., Thurmon, J. C., Grimm, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Lumb and Jones' veterinary anesthesia and analgesia. , 23-86 (2013).</li><li>Matsuda, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+newly+developed+inhalation+anesthesia+system+and+intraperitoneal+anesthesia+on+the+hemodynamic+state+in+mice.">Comparison of newly developed inhalation anesthesia system and intraperitoneal anesthesia on the hemodynamic state in mice.</a> Biological and Pharmaceutical Bulletin. 30 (9), 1716-1720 (2007).</li><li>Garber, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. 8th edn. , (2011).</li><li>Zarndt, B. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+far-infrared+active+warming+device+in+Guinea+pigs+(Cavia+porcellus).">Use of a far-infrared active warming device in Guinea pigs (Cavia porcellus).</a> Journal of the American Association for Laboratory Animal Science. 54 (6), 779-782 (2015).</li><li>Wolforth, J., Dyson, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flushing+induction+chambers+used+for+rodent+anesthesia+to+reduce+waste+anesthetic+gas.">Flushing induction chambers used for rodent anesthesia to reduce waste anesthetic gas.</a> Lab Animal. 40 (3), 76-83 (2011).</li></ol>

这个项目得到了肯特科学公司的设备和资金的支持。作者克里斯塔·比贾雷利和戴夫·菲茨米勒是肯特科学公司的员工，该公司生产本文中使用的设备。本文的开放访问出版物由肯特科学公司赞助。

这种数字低流量麻醉系统将麻醉、通风、发热和生理监测系统集成到单个设备中。此外，该系统还包含一个内部泵，允许它吸收环境空气作为载体气体，从而无需压缩气体来源。
在此过程中，该系统被用作唯一的设备，以取代麻醉蒸发器、机械呼吸机、脉冲氧化计和加热垫。我们之前演示过以100mL/min2的流速进行麻醉分娩。流速设置对于这种麻醉输送技术至关重要，因为流速直接控制使用的液体麻醉剂的体积。我们以前也演示过如何使用低流量来节省麻醉液1，2。当传统的蒸发器连接到机械呼吸机时，蒸发器必须连续运行，而呼吸机从气流中抽取样品。在带集成呼吸机的数字蒸发器中，只有通风机输出的通风所需的气体。这降低了与麻醉液、载体气体和木炭过滤器相关的成本。
虽然使用低流量数字蒸发器有很多优点，但也有局限性。该系统设计为以低流速运行，非常适合啮齿动物和其他小型哺乳动物，但不会提供高于1000 mL/min流量的麻醉。因此，这种特殊的系统只适合小型动物物种。集成脉冲氧化计仅包括用于爪子的传感器。不建议在尾部使用传感器，这可能是某些外科手术的限制。此外，虽然可以通过这个系统通过爪子传感器监测呼吸速率，但很难在较长时间内获得一致的呼吸记录。最后，与传统的蒸发器不同，这种数字系统需要电力。电池可用于无法供电或停电的情况下，并且可以通过几个小时的使用为系统供电。
此设置和协议证明安全有效地使用数字低流量麻醉系统与集成呼吸机和生理监测模块。这种设置对于任何长凳空间有限的实验室或在手术场附近存放多件设备和管子不可行的地方都是有用的。一体式系统有许多好处，包括消除压缩气箱和单独的生理监测设备。总体而言，使用传统蒸发器不理想的团体可以考虑这一集成系统。

涉及动物模型的研究往往需要专门的数据收集设备。有两种常见的麻醉蒸发器，通常用于小型动物手术。传统的麻醉蒸发器依靠基于大气压力和气体流动1，2，3，4，5，6，7，8，9，10的挥发性麻醉剂的被动蒸发。它们设计为以0.5升/分钟至10升/分钟的流速运行，是大型动物模型11的理想之选。
与传统的蒸发器12、13相比，我们最近展示了低流量数字蒸发器的效果。低流量的数字麻醉系统可用于保持动物在鼻锥上的极低流速1.5-2.2倍的动物的分钟体积14，15，16。
使用数字麻醉系统有许多好处。它集成了一个内置泵，它吸收环境空气作为载体气体。这允许用户在不使用压缩气体的情况下进行麻醉。最近的研究17，18表明，使用空气代替氧气作为载体气体可能有利于许多程序。
生理监测和加热功能也可以安装到数字低流量麻醉系统中。在大多数机构中，动物变暖和生理监测需要机构动物护理和使用委员会19、20、21、22。比较麻醉剂的生理影响的研究表明，体温、心脏功能和呼吸功能23、24、25的剧烈抑郁。通常需要将动物放在加热垫上以监测和维持正常的体温。有许多动物变暖的方法，如热水器，电加热垫和热灯，但每一个都有显著的缺点。在比较不同方法的动物变暖的研究中，远红外变暖被认为是最有益的26。数字蒸发器包括内置的家热远红外变暖，以保持特定的动物体温。这消除了任何额外的加热垫控制器的需求。
除了监测体温外，脉搏氧化是监测动物心率和氧饱和度的常用方法。这种非侵入性方法简单、准确，可全面评估动物调节血液氧分水平的能力。脉冲氧化学的爪子传感器可以连接到麻醉系统，正如我们之前所证明的2。
当动物处于较长的麻醉期或需要控制动物呼吸模式时，通常需要机械通风。低流量数字蒸发器具有在压力或体积控制下提供可控呼吸的能力。集成式呼吸机无需外部呼吸机和过多的管子设置要求。
由于所有这些常见的显示器和功能都组合成一个设备，因此油管设置大大简化。此协议的目的是演示一体式数字麻醉系统的设置和使用。

<table>
	<tbody>
		<tr>
			<td>Intubation Kit</td>
			<td>Kent Scientific Corporation</td>
			<td>ETM-MSE</td>
			<td>Includes intubation stage, intubation tube, LED light</td>
		</tr>
		<tr>
			<td>Isoflurane Liquid Inhalation 99.9%</td>
			<td>Henry Schein, Inc.</td>
			<td>1182097</td>
			<td>Glass bottle 250mL</td>
		</tr>
		<tr>
			<td>MouseSTAT Pulse Oximeter</td>
			<td>Kent Scientific Corporation</td>
			<td>SS-03</td>
			<td>Integrated into SomnoSuite</td>
		</tr>
		<tr>
			<td>Oxygen Tank</td>
			<td>Indiana Oxygen Company</td>
			<td>23-160246</td>
			<td>Medical Grade O2 99%</td>
		</tr>
		<tr>
			<td>RoVent Automatic Ventilator</td>
			<td>Kent Scientific Corporation</td>
			<td>SS-04</td>
			<td>Integrated into SomnoSuite</td>
		</tr>
		<tr>
			<td>SomnoSuite Low Flow Digital Anesthesia System</td>
			<td>Kent Scientific Corporation</td>
			<td>SS-01</td>
			<td>Includes RightTemp Homeothermic Warming control, pad, and temperature sensors</td>
		</tr>
		<tr>
			<td>SomnoSuite Mouse Starter Kit</td>
			<td>Kent Scientific Corporation</td>
			<td>SOMNO-MSEKIT</td>
			<td>Includes nose cone, syringes, induction chamber, and charcoal canister</td>
		</tr>
	</tbody>
</table>

use of an integrated low-flow anesthetic vaporizer, ventilator, and physiological monitoring system for rodents

低流量数字蒸发器通常使用注射器泵将挥发性麻醉剂直接插入载体气体流中。根据动物福利建议，动物在需要麻醉的程序中受到温暖和监测。常见的麻醉和生理监测设备包括煤气罐、麻醉蒸发器和支架、加热控制器和垫子、机械呼吸机和脉冲氧化计。计算机对于数据收集和运行设备软件也是必要的。在较小的空间或进行现场作业时，在有限的空间中配置所有这些设备可能具有挑战性。
该协议的目的是展示使用低流量数字蒸发器使用压缩氧气和室内空气的最佳做法，以及集成的机械呼吸机、脉冲氧化计和远红外加热，作为适合啮齿动物的全包麻醉和生理监测套件。

10周大，雄性，野生类型C57Bl6j小鼠体重25.41±0.8克用于这项研究。小鼠在鼻锥或插管上麻醉和维持，并在综合机械呼吸机上保持1.5-2.5%的异氟，同时监测心率和氧饱和度。这些动物被集体安置在微隔离箱中，并可免费获得标准啮齿动物和瓶装水。
心率和SpO2在维护过程中通过脉搏氧化（图5，图6和图7）进行监测。通过红外加热垫和热灯将体温维持在36.5-37.5°C。通风动物在插管过程中通过集成鼻锥的插管支架连续获得异氟兰的分娩。每只小鼠在鼻锥上成功通风或保持低流量，室空气（RA）或氧气（O2）不超过141毫升/分钟，持续15分钟。动物的心率和血氧饱和度保持稳定，所有组的测量几乎没有显著变化。SpO2在所有组保持在 82-99% 之间，而体温保持在 36.5-37.5 °C 之间。 我们观察到脉冲轴测仪的位置和体温都影响了SpO2的测量结果。如果我们从脉冲轴仪中观察到无效读数，我们调整了传感器的位置和加热水平，以保持核心体温稳定。
进行了双向 ANOVA 与 Bonferroni 校正，以确定 图 5、图 6 和 图 7中数据的重要性。低于0.05的 p值被认为是显著的。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61311/61311fig01.jpg"> 图1:麻醉感应和鼻锥维护管设置图。<a href="https://www.jove.com/files/ftp_upload/61311/61311fig01large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/61311/61311fig02.jpg"> 图2:麻醉诱导、灌输和通风管设置图。<a href="https://www.jove.com/files/ftp_upload/61311/61311fig02large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61311/61311fig03.jpg"> 图3:小鼠在插管过程中通过集成鼻锥的插管支架连续获得异氟兰。<a href="https://www.jove.com/files/ftp_upload/61311/61311fig03large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/61311/61311fig04.jpg"> 图4:集成脉冲氧化计传感器放置在后爪上。<a href="https://www.jove.com/files/ftp_upload/61311/61311fig04large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/61311/61311fig05.jpg"> 图5:平均心率超过15分钟±SD与房间空气（RA）或100%氧气（O2）通过鼻锥或通风通过气管（n=5/组）。没有发现不同组之间的显著差异。 <a href="https://www.jove.com/files/ftp_upload/61311/61311fig05large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/61311/61311fig06.jpg"> 图 6:通过低流量麻醉系统进行初始麻醉后记录的心率值 （bpm）。平均心率值在 15 分钟内从 30 秒的时间间隔计算。每个数据点表示每个组中所有动物的平均± SD （n=5）。在15分钟内，没有观察到任何组的心率有显著变化。 <a href="https://www.jove.com/files/ftp_upload/61311/61311fig06large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/61311/61311fig07.jpg"> 图7:组织氧饱和度水平（%）在用低流量麻醉系统进行初始麻醉后。平均 SpO2 值在 15 分钟内从 30 秒的时间间隔计算。每个数据点表示每个组中所有动物的平均± SD （n=5）。在15分钟内，在任何组中均未观察到SpO2 的显著变化。 <a href="https://www.jove.com/files/ftp_upload/61311/61311fig07large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>

Watch this Scientific Journal Video about Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents at JoVE.com

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

在这里，我们提出了一个协议，使用数字低流量麻醉系统与集成呼吸机和生理监测模块安全有效地管理麻醉气体给小鼠。

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 ...

use-an-integrated-low-flow-anesthetic-vaporizer-ventilator

Research

JoVE Journal

Medicine

5.8K 次观看.  Kent Scientific Corporation. 在这里，我们提出了一个协议，使用数字低流量麻醉系统与集成呼吸机和生理监测模块安全有效地管理麻醉气体给小鼠。

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

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 ...

科研

医学

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

的的膀胱测压小老鼠:A膀胱Chemosensation研究的使用

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

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 ...

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

Parkinson's disease (PD) is the second most common movement disorder and affects 1% of people over the age of 60 1. Because ageing is the most important risk factor, cases of PD will increase during the next decades 2. Next to pathological protein folding and impaired protein degradation pathways, alterations of mitochondrial function and morphology were pointed out as further hallmark of neurodegeneration in PD 3-11.
After years of research in murine and human cancer cells as in vitro models to dissect molecular pathways of Parkinsonism, the use of human fibroblasts from patients and appropriate controls as ex vivo models has become a valuable research tool, if potential caveats are considered. Other than immortalized, rather artificial cell models, primary fibroblasts from patients carrying disease-associated mutations apparently reflect important pathological features of the human disease.
Here we delineate the procedure of taking skin biopsies, culturing human fibroblasts and using detailed protocols for essential microscopic techniques to define mitochondrial phenotypes. These were used to investigate different features associated with PD that are relevant to mitochondrial function and dynamics. Ex vivo, mitochondria can be analyzed in terms of their function, morphology, colocalization with lysosomes (the organelles degrading dysfunctional mitochondria) and degradation via the lysosomal pathway. These phenotypes are highly relevant for the identification of early signs of PD and may precede clinical motor symptoms in human disease-gene carriers. Hence, the assays presented here can be utilized as valuable tools to identify pathological features of neurodegeneration and help to define new therapeutic strategies in PD.

Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes.

利用人类纤维原细胞线粒体表型的监测领域的帕金森氏病

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

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.

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 Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity (AIDN): A Translational Neuroscience Approach

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children. Recent investigations in animals have raised concerns that anesthesia exposure may lead to neuronal apoptosis, known as anesthesia-induced developmental neurotoxicity (AIDN). Furthermore, some clinical studies in children have suggested that anesthesia exposure may lead to neurodevelopmental deficits later in life. Nonetheless, an ideal animal model for preclinical study has yet to be developed. The neonatal piglet represents a valuable model for preclinical study, as they share a striking number of developmental similarities with humans.
The anatomy and physiology of piglets allow for implementation of rigorous human perioperative conditions in both survival and non-survival procedures. Femoral artery catheterization allows for close monitoring, thus enabling prompt correction of any deviation of the piglet's vital signs and chemistries. In addition, there are multiple developmental similarities between piglets and human neonates. The techniques required to use piglets for experimentation will require experience to master. A pediatric anesthesiologist is a critical member of the investigative team. We describe, in a general sense, the appropriate use of a piglet model for neurodevelopmental study.

Use of a Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity AIDN: A Translational Neuroscience Approach

Anesthesia-induced developmental neurotoxicity (AIDN) research has focused on rodents, which are not broadly applicable to humans. Non-human primate models are more relevant, but are cost-prohibitive and difficult to use for experimentation. The piglet, in contrast, is a clinically relevant, practical animal model ideal for the study of anesthetic neurotoxicity.

使用仔猪模型研究麻醉诱发的发育神经毒性（AIDN）:一种转化神经科学方法

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children.  Recent investigations in animals have raised concerns that ...

Sucrose Preference and Novelty-Induced Hypophagia Tests in Rats using an Automated Food Intake Monitoring System

The prevalence and incidence of depressive disorders are rising worldwide, affecting about 322 million individuals, underlining the need for behavioral studies in animal models. In this protocol, to study depression-like and anhedonic behavior in rats, the established sucrose preference and novelty-induced hypophagia tests are combined with an automated food and liquid intake monitoring system. Prior to testing, in the sucrose preference paradigm, male rats are trained for at least 2 days to consume a sucrose solution in addition to tap water. During the test, rats are again exposed to water and sucrose solution. Consumption is registered every second by the automated system. The ratio of sucrose to total water intake (sucrose preference ratio) is a surrogate parameter for anhedonia. In the novelty-induced hypophagia test, male rats undergo a training period in which they are exposed to a palatable snack. During training, rodents show a stable baseline snack intake. On test day, the animals are transferred from home cages into a fresh, empty cage representing a novel unknown environment with access to the known palatable snack. The automated system records the total intake and its underlying microstructure (e.g., latency to approaching the snack), providing insight into anhedonic and anxious behaviors. The combination of these paradigms with an automated measuring system provides more detailed information, along with higher accuracy by reducing measuring errors. However, the tests use surrogate parameters and only depict depression and anhedonia in an indirect manner.

Presented here is a protocol to study depression-like and anhedonic behavior in rats. It combines two well-established behavioral methods, the sucrose preference and novelty-induced hypophagia tests, with an automated food and liquid intake monitoring system, to indirectly investigate rodent behavior using surrogate parameters.

使用自动食物摄入量监测系统对大鼠的蔗糖偏好和新奇诱导性低吞咽测试

The prevalence and incidence of depressive disorders are rising worldwide, affecting about 322 million individuals, underlining the need for behavioral ...

TBase - an Integrated Electronic Health Record and Research Database for Kidney Transplant Recipients

TBase is an electronic health record (EHR) for kidney transplant recipients (KTR) combining automated data entry of key clinical data (e.g., laboratory values, medical reports, radiology and pathology data) via standardized interfaces with manual data entry during routine treatment (e.g., clinical notes, medication list, and transplantation data). By this means, a comprehensive database for KTR is created with benefits for routine clinical care and research. It enables both easy everyday clinical use and quick access for research questions with highest data quality. This is achieved by the concept of data validation in clinical routine in which clinical users and patients have to rely on correct data for treatment and medication plans and thereby validate and correct the clinical data in their daily practice. This EHR is tailored for the needs of transplant outpatient care and proved its clinical utility for more than 20 years at Charit&#233; - Universit&#228;tsmedizin Berlin. It facilitates efficient routine work with well-structured, comprehensive long-term data and allows their easy use for clinical research. To this point, its functionality covers automated transmission of routine data via standardized interfaces from different hospital information systems, availability of transplant-specific data, a medication list with an integrated check for drug-drug interactions, and semi-automated generation of medical reports among others. Key elements of the latest reengineering are a robust privacy-by-design concept, modularity, and hence portability into other clinical contexts as well as usability and platform independence enabled by HTML5 (Hypertext Markup Language) based responsive web design. This allows fast and easy scalability into other disease areas and other university hospitals. The comprehensive long-term datasets are the basis for the investigation of Machine Learning algorithms, and the modular structure allows to rapidly implement these into clinical care. Patient reported data and telemedicine services are integrated into TBase in order to meet future needs of the patients. These novel features aim to improve clinical care as well as to create new research options and therapeutic interventions.

TBase combines an electronic health record with an innovative research database for kidney transplant recipients. TBase is built upon an in-memory database platform, connected to different hospital systems, and used for regular outpatient care. It automatically integrates all relevant clinical data including transplantation-specific data creating a unique research database.

TBase - 肾移植受者的综合电子健康记录和研究数据库

TBase is an electronic health record (EHR) for kidney transplant recipients (KTR) combining automated data entry of key clinical data (e.g., laboratory ...

Multi-system Monitoring for Identification of Seizures, Arrhythmias and Apnea in Conscious Restrained Rabbits

Patients with ion channelopathies are at a high risk of developing seizures and fatal cardiac arrhythmias. There is a higher prevalence of heart disease and arrhythmias in people with epilepsy (i.e., epileptic heart.) Additionally, cardiac and autonomic disturbances have been reported surrounding seizures. 1:1,000 epilepsy patients/year die of sudden unexpected death in epilepsy (SUDEP). The mechanisms for SUDEP remain incompletely understood. Electroencephalograms (EEG) and electrocardiograms (ECG) are two techniques routinely used in the clinical setting to detect and study the substrates/triggers for seizures and arrhythmias. While many studies and descriptions of this methodology are in rodents, their cardiac electrical activity differs significantly from humans. This article provides a description of a non-invasive method for recording simultaneous video-EEG-ECG-oximetry-capnography in conscious rabbits. As cardiac electrical function is similar in rabbits and humans, rabbits provide an excellent model of translational diagnostic and therapeutic studies. In addition to outlining the methodology for data acquisition, we discuss the analytical approaches for examining neuro-cardiac electrical function and pathology in rabbits. This includes arrhythmia detection, spectral analysis of EEG and a seizure scale developed for restrained rabbits.

Using simultaneous video-EEG-ECG-oximetry-capnography, we developed a methodology to evaluate the susceptibility of rabbit models to develop provoked arrhythmias and seizures. This novel recording system establishes a platform to test the efficacy and safety of therapeutics and can capture the complex cascade of multi-system events that culminate in sudden death.

意识约束兔子中癫痫发作、心律失常和呼吸暂停的多系统监测

Patients with ion channelopathies are at a high risk of developing seizures and fatal cardiac arrhythmias. There is a higher prevalence of heart disease ...

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter

As a key event of cardiovascular system diseases, coronary artery disease (CAD) has been widely regarded as the main culprit of atherosclerosis, myocardial infarction, and angina pectoris, which seriously threaten the life and health of people all over the world. However, how to record the dynamic biomechanical characteristics of isolated blood vessels has long puzzled people. Meanwhile, precise positioning and isolation of coronary arteries to measure in vitro dynamic vascular tension changes have become a trend in CAD drug development. The present protocol describes the macroscopic identification and microscopic separation of rat coronary arteries. The contraction and dilation function of the coronary artery ring along the vessel diameter was monitored using the established multi myograph system. The standardized and programmed protocols of coronary ring tension measurement, from sampling to data acquisition, tremendously improve the repeatability of the experimental data, which ensures the authenticity of vascular tension records after physiological, pathological, and drug intervention.

The present protocol describes the wire myograph technique for measuring vascular reactivity of the rat coronary artery.