作者希望在学习这种插管技术的过程中, 对参加过兔子处理和麻醉训练课程的员工和研究人员进行反馈。

下面概述的插管程序由梅奥诊所机构动物护理和使用委员会批准. 
 1. 准备工作 ( 图 1 ) <ol> <li> 获得5法国聚丙烯导管并切断 non-rounded 端, 因此总长度为8-10 英寸。标记油管的切口端, 以清楚地指示应将哪个端 不 插入气管 (箭头, 图 1 )。
	<li> 在喉镜上放置一个大小为0或1的米勒喉镜刀片, 并验证指示灯是否正常工作. </li>
	<li> 将0.25 毫升2% 利多卡因注射到1毫升注射器中. </li>
	<li> 在适当大小的 uncuffed 气管插管端使用无菌润滑剂 (通常为 2.5-4.0 mm). </li>
	<li> 准备在插入后固定气管插管的用品 ( 例如, 脐带胶带). </li>
	<li> 获取在程序结束时检测气流的设备, 以确认成功的插管 ( #5, 图 1 )。这是可选的. </li>
</ol> 2。麻醉 <ol> <li> 管理注射麻醉, 如35毫克/千克氯胺酮和5毫克/千克嗪注使用3毫升注射器与25测针在四头肌的后腿的约4月大, 3.0-3.5 公斤, 新西兰白色兔. 
	注: 这种麻醉方案是一种常用的注射剂组合在年轻, 健康, 特定的无病原体研究兔, 但根据临床情况和/或实验目标, 不同的协议可能更合适. </li>
	<li> 在兔被麻醉后, 将无菌的眼部润滑剂应用于眼睛以保护角膜组织. </li>
</ol> 3。气管插管 <ol> <li> 测量从门牙到胸廓入口的气管导管, 确定插入气管导管的适当长度。在胸廓入口的水平处放置管末端, 可以确保导管的开度超过喉部, 但颅至气管分岔处. </li>
	<li> 将兔 sternally 放在准备表上, 将其头部略微延伸到表的边缘 ( 图 2 )。应将兔的头部和脊柱置于正方形的直线上. </li>
	<li> 抬起头, 用纱布垫把兔子和 #39 的舌头侧向兔子和 #39; 右下门牙 (在纵), 注意避免门牙的外伤. </li>
	<li> 用右手握住喉镜和刀片, 然后把它放进门牙后面的嘴里, 从兔子和 #39 的左边进入. </li>
	<li> 沿尾端向口的顶部, 直到软腭可见. </li>
	<li> 用右手, 将喉镜向前 (弯曲手腕, 按住喉镜向下), 同时延长兔和 #39; s 头向后和颈部向前, 以保持一个开放的呼吸道和查看喉 ( 图 3 A ). 作为强制鼻呼吸, 会厌常高于软腭, 使用这种方法压缩舌头的底座以取代会厌并暴露声门。为了防止氧气饱和, 重要的是保持颈部延长, 以保持一个开放的气道在插管. </li>
	<li> 松开舌, 并将喉镜刀片切换到左手, 释放右手以放置导导管. </li>
	<li> 在持有喉镜和保持声门的视图时 ( 图 3 B ), 通过通过喉部的开口 ( 图 4 ) 推进导管的圆尖).导管会在气管分岔处自然停止. </li>
	<li> 在保持引导导管的同时, 抓住气管导管, 并将其穿线在导尿管上, 注意不要在气管导管连接器上抓住它的力量。推进管, 直到有阻力从声带 ( 图 5 ). </li>
	<li> 将0.25 毫升2% 利多卡因注入气管导管的开口。利多卡因会沿着气管插管到局部麻醉声带, 防止痉挛 22 ( 图 6 )。
	<li> 允许大约两分钟的局部麻醉生效, 并通过轻轻地扭转气管插管, 推进它通过声带 ( 图 7 )。一旦管子通过声带, 取下喉镜和导管, 注意不要把管子取出. </li>
	<li> 通过观察气流来验证气管插管的位置。有多种方法来实现这一点, 包括放置的组织或毛皮在年底的管, 以检测气流, 寻找雾的管或雾的玻璃/镜子在年底的管, 并听取气流。设计用于检测气流 23 或呼吸的设备也可以使用 ( 图 8 ) 来确定插管是否成功 (请参阅 材料表 选项)。使用优选的方法 (如脐带绑在管和头上) 固定气管插管。在呼吸时, 将麻醉回路和 auscult 两侧的胸腔连接起来, 以保证肺部两侧的通气. </li>
</ol>

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作者没有什么可透露的。

在这里, 我们提供了一个兔子插管技术的详细描述, 可以很容易地掌握在任何临床设置。此技术类似于以前发布的直接可视化技术10,152025 , 还有一些额外的改进, 包括使用 3.5 mm uncuffed 气管插管和5法国导尿管作为我们选择的指南, 已被切断, 并在削减结束标记与永久标志。兔气管容易受伤, 因此, uncuffed 气管导管的使用通常建议当插管兔, 以防止损伤的气管粘膜从压力的袖口。26我们选择的5法国导尿管在喉部直接可视化过程中, 小到可以轻松地通过声带, 而不会阻碍视野。虽然小, 这个导管是僵硬的, 足以保持在地方作为指导的气管插管。远端的导管是四舍五入, 以帮助减少创伤时, 分歧达到。导管的近端被切断, 使导管在一个可行的长度 (不太长) 和删除的结束, 扩大为附件的注射器。切口末端标记以永久标记作为视觉指示不插入此末端入气管, 可能导致损伤对气管黏膜。
这项技术的主要限制是, 因为它确实需要使用喉镜刀片, 它可能是不可行的非常年幼的兔子或非常小的品种, 由于较小的嘴巴大小。此外, 一些设施可能没有这种规模的喉镜刀片现成的可用。在作者的机构, 这种方法是用于新西兰白 (≥6周) 和成年荷兰兔, 但我们没有尝试这种方法在年轻的动物或较小的品种。在兔子的病人很小的情况下, 或者如果一个喉镜与一个小米勒刀片是不可用的, 另一种选择是使用手持镜而不是喉镜。15,20使用带有 #5 耳窥器的镜, 将喉部可视化, 引导导管由直接可视显示, 并在按照上述步骤进行操作之前将窥镜移除。
当导管的末端进入气管并触及气管的分岔时, 使用导导管来避免造成外伤或支气管的可能性是非常重要的。这些并发症的风险可以减少或避免, 通过喂养导管与非常温和的动作, 一旦有阻力停止, 并避免头部和颈部的移动一旦导管到位。此外, 当在导管上推进气管插管时, 导管连接器上也有可能会发现导尿管。因此, 重要的是要确保导管通过连接器滑动而不捕获;或者, 连接器可以在固定气管插管或附着在麻醉回路时被移除和更换。
有任何插管方法, 有潜在的并发症和不良反应。使用喉镜直接可视化方法, 不正确插入喉镜刀片或积极放置气管插管可导致喉、舌或牙齿损伤。正确定位的动物是第一个关键的步骤, 最大限度地减少创伤的可视化喉。脊柱应伸直, 头部与脊柱直接对准;如果动物不是直的, 或者如果颈部被扭到左边或右边, 这将使喉部的可视化更加困难, 并可能导致喉镜刀片的过度创伤。使用这项技术成功的另一个关键关键是沿着口的顶部朝向喉部, 一旦软腭被可视化, 就会通过弯曲手腕将刀片移到适当的位置。这保持头到位, 而移动刀片的尖端到最佳的观察位置声带, 位于相当腹在兔子20,24。
由于直接可视化喉可能是棘手的, 如果精确的技术, 我们上面概述没有遵循, 使用替代插管策略, 不需要可视化喉可以吸引。例如, 盲技术是一种可能的选项, 不需要使用喉镜16,17,18,19,20。然而, 这项技术的使用依赖于通过气管插管检测气流的能力, 以便引导导管的位置, 如果动物的呼吸非常浅或被逮捕, 使用这种方法是不可靠的。声门设备也可以考虑, 因为放置这些设备不需要显着的喉, 他们通常很容易到位。这些设备不直接进入气管, 然而, 因此有潜在的缺乏直接流到气道, 无法通风, 如果痉挛发展, 缺乏密封足够的通风在开程序或紧急情况胃音的复苏和发展3,4,8。专门为兔子设计的新设备可能提供了一个很有前途的替代品, 但是需要更多的研究来评估这些设备, 而对于缺乏进入气管的担忧仍然存在。胃音也可能是一个潜在的并发症, 在插管动物, 如果气管插管是意外放置在食道;然而, 直接可视化技术的使用, 通过验证引导导管正确放置到喉部, 从而最大限度地减少了这种并发症的风险。
杓和/或气管粘膜的创伤是另一个潜在的问题, 当插入气管插管, 特别是当使用充气的手铐管26,27,28。家兔气管易受损伤, 可能导致皮下气肿、气胸等并发症, 甚至死亡10,27,28,29。在执行我们在这里描述的技术时, 应该注意预防气管损伤, 使用 uncuffed 气管插管, 确保引导导管的圆形尖端插入气管, 而不是将导导管强行注入声门。此外, 选择一个适当大小的气管插管是至关重要的。气管导管大小应考虑与动物的大小, 以及与开放的喉口, 因为它比气管直径小。气管导管直径过大可能导致杓粘膜或软骨的损伤。另一方面, 使用较小的气管导管会导致呼吸道阻力增加, 并增加因粘液或扭结而阻塞的几率。因此, 呼吸道监测在插管家兔中是非常重要的。除了脉冲氧, 气流的测量应 monitored 与二氧化碳;如果分析仪不可用, 气道监测器是一种可用于检测空气流量的替代品 (但不提供呼出的二氧化碳的测量)。此外, 提供辅助通风是有价值的, 特别是在使用较小直径管的情况下24,30。
除了已经提到的使用这项技术的好处之外, 这种方法的另一个主要优点是它可以由一个单独的个体来执行, 而无需助手, 这在繁忙的临床环境中可能是很重要的。在这种情况下, 重要的是要收集所有必要的用品, 并有他们在容易达到的插管过程中, 特别是当引导导管到位, 并应避免头部的移动。
我们在这里描述的技术是由我们机构的兽医和研究人员使用的所有兔插管, 这相当于每年大约200-400 插管。这是很容易教在一个典型的兔麻醉训练课程, 这需要大约30分钟。我们发现, 它是相对容易学习, 与学生学习它在他们的前3尝试, 一般能够成功地重复的过程中, 在下一个额外的尝试。在将来的研究中, 与其他插管技术相比, 将时间与熟练程度和保持熟练程度的能力进行比较是有价值的。虽然研究了盲技术的变体16, 但尚未对各种插管技术的学习曲线进行全面评估。另一个潜在的未来研究方向可能包括比较直接气管插管到较新的 species-specific 声门气道装置, 特别是在正压通气期间的功能。
总之, 我们在这里描述的家兔插管技术是无创的, 可靠的, 不需要使用专门的设备, 易于掌握, 并且可以由一个单独的个体来完成, 无需助手。所有的呼吸道接入技术都存在一定程度的风险, 但是通过直接的可视化技术, 并按照我们上面描述的步骤来减少并发症, 这些风险远远超过了在麻醉期间有足够气道的好处。

家兔有一个狭窄的, 深的口腔, 限制了气管插管的喉的能见度。它们还有非常大的肉质舌和颊齿, 当与狭窄的口腔结合时, 在插入喉镜和可视声门插入气管插管1时会引起问题。在尝试插管时, 必须非常小心避免造成口腔和呼吸道的损伤。喉组织是非常易碎和过度的创伤从反复尝试插管可能导致明显的炎症, 使插管更加困难2。
由于这些原因, 各种替代技术已经被描述, 特别是声门气道装置, 包括喉管3,4, 喉罩气道 (LMA)4,5,6和 species-specific 声门气道设备7。喉管包括一个管子与二个气球直接气流入气管: 一个更大的气球在管子中间形成封印在上部咽和一个更小的气球在远端封印食道入口3。该 LMA 由一个椭圆形的管, 在远端, 充气形成一个密封的密封在声门4,5,6。species-specific 装置是第一个兽医 species-specific 声门气道装置, 目前可用于猫和兔, 旨在适合每个物种的解剖结构, 以创建一个压力密封在咽7。然而, 由于这些设备不直接进入气管, 它们可能会出现诸如胃音等并发症的风险。气管内缺乏适当的密封, 这就引起了人们对使用这些装置用于呼吸支持、急诊或胸部手术的正压通气是否足够的担忧3,8。此外, 缺乏直接插管的气管可能导致人员暴露于废麻醉剂气体5。
许多方法的直接气管插管已被描述9,10,11,12,13,14,15。一种常用的方法是盲法, 因为喉镜的插入不需要16,17,18,19,20. 作为一种视觉盲技术, 该方法需要通过气管插管进行空气流动监测, 并进行细致的操作以引导放置。虽然这种方法是常用的研究和临床兽医实践, 在作者的经验, 这项技术可能是费时, 并不总是成功的, 因此更难教, 并可能需要额外剂量的麻醉药物9,10. 此外, 由于监测呼吸是用来指导适当的放置管, 这种方法将是更困难和不可靠的呼吸骤停。另一种流行的方法是使用传统的喉镜技术来插管, 就像做了较大的物种。然而, 传统的喉镜技术在家兔是困难的, 因为声带的看法是阻塞时, 气管插管插入到嘴里。因此, 使用这种方法可以导致反复插管尝试, 增加麻醉剂量, 并伤害口腔和气道10。
更昂贵和技术上具有挑战性的选择插管也存在。纤维喉镜可以方便地插入气管插管, 将喉镜插入管内, 并在屏幕上可视化插入气管。同样, 内窥镜可用于可视化喉区和引导放置的管。然而, 这两种方法都需要使用昂贵的设备, 在典型的临床设置9、1112中可能无法轻易获得。通过鼻腔插管是一种替代方法, 已被描述, 虽然这是一个风险引入机会病原体从鼻腔进入肺部, 特别是在兔子, 不是特定的-无病原体的21。最后, 更有侵入性的插管方法也已公布, 包括逆行插入导管通过针放置经到气管, 以及使用针 cricothyroidotomy 后, 引导导线的位置,容器扩张和鞘介绍人13,14。作为侵入性技术, 这些方法有潜在的并发症, 如 post-procedural 疼痛和感染。我们在这里描述的技术, 使用喉镜和导管, 可以教与短时间的训练, 以熟练, 是无创的, 可以执行由一个单一的个人不需要一个助手, 不需要使用专用设备。

<table><tbody><tr><td>Ketalar&lt;br/&gt; Ketamine HCl Inj.</td><td>JHP Pharmaceuticals</td><td>NDC 42023-113-10</td><td>35mg/kg IM in epaxial muscles or quadriceps muscle (preferred)</td></tr><tr><td>Anased&lt;br/&gt; Xylazine (100mg/ml)</td><td>Lloyd Labs</td><td>NADA #139-236</td><td>5mg/kg IM in epaxial muscles or quadriceps muscle (preferred)</td></tr><tr><td>Welch Allyn Laryngoscope Handle</td><td>Welch Allyn</td><td>60300</td><td>Requires 2 size C alkaline batteries</td></tr><tr><td>Size 0 or 1 Miller Laryngoscope Blade</td><td>Welch Allyn</td><td>68040</td><td></td></tr><tr><td>2.5 &amp;ndash; 4.0 mm ID uncuffed endotracheal tube</td><td>Jorgenson</td><td>J0149V</td><td>Suitable for 2.5-5 kg rabbits&lt;br/&gt; 3.5 mm most commonly used</td></tr><tr><td>Sovereign polypropylene catheter, 5 Fr x 22&amp;rdquo;</td><td>Medtronic</td><td>8890703310</td><td>Cut to 8-10 inches&lt;br/&gt; Mark the cut end with permanent marker to indicate which end not to insert</td></tr><tr><td>Lidocaine 2% HCL Inj</td><td>VetOne</td><td>L-2000-04</td><td></td></tr><tr><td>4-Ply Nonsterile, Bulk Packed Versalon Sponges (Gauze pads)</td><td>Covidien</td><td>KC9024A</td><td></td></tr><tr><td>Tape, Twill, Bleached &amp;frac12;&amp;rdquo; x 72 yds</td><td>Encompass Textiles</td><td>48210-025</td><td>Used for securing endotracheal tube</td></tr><tr><td>HR Lubricating Jelly</td><td>HR Pharmaceuticals</td><td>201</td><td>For lubricating endotracheal tube tip</td></tr><tr><td>Great Plains Ballistics: BAAM (Beck Airway Airflow Monitor) 22</td><td>BoundTree Medical</td><td>21410</td><td>For confirmation of successful intubation</td></tr><tr><td>Ambu Pediatric Co2 Detector</td><td>AMBU INC</td><td>172713</td><td>For confirmation of successful intubation</td></tr></tbody></table>

endotracheal intubation of rabbits using a polypropylene guide catheter

由于其解剖异常, 家兔的气管插管可能具有挑战性。在麻醉期间实现一个专利气道对于避免气道阻塞、预防胃音和允许通气支持至关重要。由于插管困难, 探讨了使用喉罩气管或喉管等替代方法。然而, 这些方法并不导致直接进入气管, 因此可能出现并发症的发展风险。此外, 缺乏直接插管的气管可能导致人员暴露于废弃的麻醉气体。许多方法的气管插管已被描述, 包括盲目放置, 使用纤维喉镜或内窥镜, 和环状软骨放置。尽管有这么多的出版物, 许多人仍在努力取得成功。在这里, 我们提供了一个插管技术的详细描述, 可以教与短时间的训练, 以熟练程度最低。简要地, 经过注射麻醉和适当的定位兔, 聚丙烯导管被放置到气管直接可视化使用喉镜。导管被用作引导气管插管进入气管的指南。 这种方法可以在不需要昂贵设备的情况下进行插管, 并且可由单个个人执行, 而无需助手。 总之, 这项技术可以很容易地教授和执行在任何临床或研究设置很少的成本。

在作者的机构, 这项技术通常教授给实验室和兽医人员在训练期间的兔子处理和麻醉技术。教学课程是用专门的训练动物进行的。培训师在培训师的指导下, 演示了技术, 然后受训者完成了这项技术。受训者通常在1-3 次尝试中取得成功。在他们成功地对兔子进行了一次插管后, 个体被要求第二次进行插管, 并且通常能够在第一次额外的尝试中再次成功地给兔子插管。这些培训期间的反馈, 特别是以前受过替代技术训练的个人, 一贯包括对这项技术如何容易学习和成功完成的评论。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56369/56369fig1.jpg"> 
图 1。用于气管插管的供应: 1) 喉镜与大小0或1米勒喉镜刀片, 2) 脐带 (或其他供应的安全气管插管), 3) 0.25 毫升2% 利多卡因, 4) 5-法国聚丙烯导管切割到8-10 英寸non-rounded 端标记有一个永久性的标志, 明确指出不应插入气管 (箭头), 5) 空气流动检测装置, 6) 无菌润滑剂, 适用于远端的管, 7) uncuffed 气管插管。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig1large.jpg" style="font-size: 14px;" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56369/56369fig2.jpg"> 
图 2: 胸骨定位用于气管插管.兔子被定位 sternally, 前肢延伸到桌子的末端。后肢向后伸展, 兔头与脊柱对齐。在准备插入喉镜时, 用左手将头部抬高至伸展。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56369/56369fig3.jpg"> 
图 3: 喉镜插入.A) 喉镜用右手插入左纵, 在口的顶部跟随刀片的尖端, 直到软腭可见。一旦软腭是可见的, 喉镜是向前倾斜, 同时延长兔的头部背部和颈部向前, 以可视化喉。B) 喉的可视化。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56369/56369fig4.jpg"> 
图 4: 导导管的推进.A) 在将喉镜刀片切换到左手后, 右手通过喉部的开口向前推进导管的圆尖。B) 导导管通过声带通过直接可视化。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56369/56369fig5.jpg"> 
图 5: 气管导管的推进.气管内管是螺纹在导导管上, 并先进, 直到有阻力从声带。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56369/56369fig6.jpg"> 
图 6: 利多卡因灌输.利多卡因 (2%、0.25 毫升) 被注入气管导管的开口。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56369/56369fig7.jpg"> 
图 7: 气管插管的进展.利多卡因注入两分钟后, 气管导管通过轻柔的扭动, 通过声带而推进。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/56369/56369fig8.jpg"> 
图 8: 通过气管插管验证气流.气流检测装置可以用来验证气管导管内的气流, 也可以使用牙科镜来检测呼吸指示成功插管的冷凝。<a href="//ecsource.jove.com/files/ftp_upload/56369/56369fig8large.jpg" style="font-size: 14px;" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Endotracheal Intubation of Rabbits Using a Polypropylene Guide Catheter at JoVE.com

Endotracheal Intubation of Rabbits Using a Polypropylene Guide Catheter

家兔的气管插管由于其解剖异常而具有挑战性。在这里, 我们提出了一种技术, 直接插管的气管使用聚丙烯导管作为指导。这种方法使用相对低廉的用品, 需要最少的培训, 并且可以很容易地在任何临床环境中执行。

用聚丙烯导管引导家兔气管插管

Endotracheal intubation in rabbits can be challenging due to their unusual anatomy. Achieving a patent airway during anesthesia is critical for the ...

endotracheal-intubation-rabbits-using-polypropylene-guide

Research

JoVE Journal

Biology

33.4K 次观看.  Mayo Clinic. 家兔的气管插管由于其解剖异常而具有挑战性。在这里, 我们提出了一种技术, 直接插管的气管使用聚丙烯导管作为指导。这种方法使用相对低廉的用品, 需要最少的培训, 并且可以很容易地在任何临床环境中执行。

视频: Endotracheal Intubation of Rabbits Using a Polypropylene Guide Catheter

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are necessary for whole animal experiments that require surgery under deep anesthesia or measurements of lung function. Tracheostomy has been the standard for intubating the airway in these mice to allow mechanical ventilation. Orotracheal intubation has been reported but has not been successfully used in many studies because of the substantial technical difficulty or a requirement for highly specialized and expensive equipment. Here we report a technique of direct laryngoscopy using an otoscope fitted with a 2.0 mm speculum and using a 20 G&#160;intravenous catheter as an endotracheal tube. We have used this technique extensively and reliably to intubate and conduct accurate assessments of lung function in mice. This technique has proven safe, with essentially no animal loss in experienced hands. Moreover, this technique can be used for repeated studies of mice in chronic models.

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

We have developed a simple, reliable, and relatively inexpensive method for endotracheal intubation in mice via direct laryngoscopy using an otoscope with a 2.0 mm speculum. This technique is atraumatic and can be used for repeated measurements in chronic experiments. We find it superior to tracheostomy or previously reported nonsurgical techniques.

气管插管小鼠<em&gt;通过</em&gt;直接喉镜使用耳镜

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are ...

科研

医学

A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated procedures to obtain such measurements in the same animal are generally not feasible.  Here, we demonstrate protocols for obtaining from mice repeated measurements of AHR and bronchoalveolar lavage fluid samples.  Mice were challenged intranasally seven times over 14 days with a potent allergen or sham treated.  Prior to the initial challenge, and within 24 hours following each intranasal challenge, the same animals were anesthetized, orally intubated and mechanically ventilated.  AHR, assessed by comparing dose response curves of respiratory system resistance (RRS) induced by increasing intravenous doses of acetylcholine (Ach) chloride between sham and allergen-challenged animals, were determined.  Afterwards, and via the same intubation, the left lung was lavaged so that differential enumeration of airway cells could be performed.  These studies reveal that repeated measurements of AHR and BAL fluid collection are possible from the same animals and that maximal airway hyperresponsiveness and airway eosinophilia are achieved within 7-10 days of initiating allergen challenge.  This novel technique significantly reduces the number of mice required for longitudinal experimentation and is applicable to diverse rodent species, disease models and airway physiology instruments.  

Repeated measurements of rodent respiratory physiology and sampling of airway inflammatory cells are desirable, but generally not feasible. Here we describe a repeatable method for orally intubating mice that permits repeated measurements of airway hyperreactivity and sampling of airway inflammatory cells.

一个可逆的，非侵入性方法，气道阻力测量和支气管肺泡灌洗小鼠流体取样

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated ...

生物学

Blind Endotracheal Intubation in Neonatal Rabbits

The newborn rabbit is a useful animal model for various pathologies and procedures. Airway management of the rabbit is complex due to its anatomical characteristics, which is further complicated in the case of the newborn. Of the different methods of advanced airway management, endotracheal intubation is less aggressive than tracheostomy, and is more feasible than supraglottic management given the lack of supraglottic devices of such a small size. As direct glottis visualization is very difficult in animals this size, this blind intubation model is presented as an effective alternative, especially for experiments requiring prolonged anesthesia. Using this method, we performed blind intubations with a 90% success rate.

We describe a technique of endotracheal intubation in newborn rabbits after esophageal catheterization with a gastric tube.

新生儿兔子的盲内窥管插管

The newborn rabbit is a useful animal model for various pathologies and procedures. Airway management of the rabbit is complex due to its anatomical ...

Simplifying Transcutaneous Intratracheal Drug Delivery in the Newborn Preterm Rabbit

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and minimizing systemic drug exposure. The transcutaneous technique is simple and allows for the IT delivery of substances to the lung of prematurely born rabbits shortly after birth. Newborn pups are anesthetized with inhaled Isoflurane before being placed in a supine position with the neck extended. The larynx is identified and stabilized before transcutaneous placement of a 26-gauge (G) catheter into the trachea. Following catheterization of the trachea, a 30 G blunt needle attached to a Hamilton syringe is introduced into the IT catheter and is used for delivering a precise volume into the trachea during spontaneous respiration. After the IT injection is completed, the needle and catheter are withdrawn, and the pup is allowed to recover from anesthesia. Transcutaneous IT injection delivers a large proportion of the injected substance to the lung, with the majority remaining in the lung 3 hours after the intervention. The injections are well tolerated from the day of birth and can be repeated for multiple consecutive days without influencing survival. This technique can be used to investigate the effect of pharmaceutical agents on lung development and in the prevention of neonatal lung injury in preterm rabbits.

<meta charset="utf8"></head><body>简化新生早产儿经皮气管内药物输送

Transcutaneous intratracheal injection allows for effective intrapulmonary drug delivery during spontaneous respiration. Single and multiple injections are well tolerated with no effect on survival. The technique is simple to perform and can examine the effect of substances on lung development and the prevention of lung injury in newborn rabbits.

简化新生早产儿经皮气管内药物输送

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and ...

Isolated Lung Perfusion System in the Rabbit Model

The isolated lung perfusion system has been widely used in pulmonary research, contributing to elucidate the lungs&#39; inner workings, both micro and macroscopically. This technique is useful in the characterization of pulmonary physiology and pathology by measuring metabolic activities and respiratory functions, including interactions between circulatory substances and the effects of inhaled or perfused substances, as in drug testing. While in vitro methods involve the slicing and culturing of tissues, the isolated ex vivo lung perfusion system allows to work with a complete functional organ making possible the study of a continuous physiological function while recreating ventilation and perfusion. However, it should be noted that the effects of the absence of central innervation and lymphatic drainage still have to be fully assessed. This protocol aims to describe the assembly of the isolated lung apparatus, followed by the surgical extraction and cannulation of lungs and heart from experimental lab animals, as well as to display the perfusion technique and signal processing of data. The average viability of the isolated lung ranges between 5-8 h; during this period, the pulmonary capillary permeability increases, causing edema and lung injury. The functionality of preserved pulmonary tissue is measured by the capillary filtration coefficient (Kfc), used to determine the extent of pulmonary edema through time.

The isolated rabbit lung preparation is a gold standard tool in pulmonary research. This publication aims to describe the technique as developed for the study of physiological and pathological mechanisms involved in airway reactivity, lung preservation, and preclinical research in lung transplantation and pulmonary edema.

兔模型中的孤立肺灌注系统

The isolated lung perfusion system has been widely used in pulmonary research, contributing to elucidate the lungs&#39; inner workings, both micro and ...

Endotracheal Intubation Using a Flexible Intubation Endoscope as a Standardized Model for Safe Airway Management in Swine

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway or high ventilation pressures. Endotracheal intubation is a challenging skill,&#160;requiring a minimum number of successful endotracheal intubations to achieve a high success rate under optimal conditions, which is often unachievable for non-anaesthesiology researchers. Due to the specific porcine airway anatomy, a difficult airway can usually be assumed. The impossibility of establishing a secure airway can result in injury, adverse events, or death of the laboratory animal. Using a prospective, randomized, controlled evaluation approach, it has been shown that fiberoptic-assisted endotracheal intubation takes longer but has a higher first-pass success rate than conventional intubation without causing clinically relevant drops in oxygen saturation. This model presents a standardized regimen for endoscopically guided endotracheal intubation, providing a secured airway, especially for researchers who are inexperienced in the technique of endotracheal intubation via direct laryngoscopy. This procedure is expected to minimize animal suffering and unnecessary animal losses.

The use of pigs in research has increased in recent years. Nevertheless, pigs are characterized by difficult airway anatomy. By demonstrating how to perform endoscopically guided endotracheal intubation, the present protocol aims to further increase laboratory animals' safety to avoid animal suffering and unnecessary death.

使用柔性插管内窥镜作为猪安全气道管理的标准化模型的气管插管

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway ...

Model of Ischemia and Reperfusion Injury in Rabbits

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and survival experiments. New Zealand White adult rabbit is sedated with atropine, acepromazine, butorphanol, and isoflurane. The animal is intubated and placed on mechanical ventilation. An intravenous catheter is inserted into the marginal ear vein for the infusion of medications. The animal is pre-medicated with heparin, lidocaine, and lactated Ringer&#39;s solution. A carotid cut-down is performed to obtain arterial line access for blood pressure monitoring. Select physiologic and mechanical parameters are monitored and recorded by continuous real-time analysis.
With the animal sedated and fully anesthetized, either a fourth intercostal space small left thoracotomy (survival) or midline sternotomy (non-survival) is performed. The pericardium is opened, and the left anterior descending (LAD) artery is located.
A polypropylene suture is passed around the second or third diagonal branch of the LAD artery, and the polypropylene filament is threaded through a small vinyl tube, forming a snare. The animal is subjected to 30 min of regional ischemia, achieved by occluding the LAD by tightening the snare. Myocardial ischemia is confirmed visually by regional cyanosis of the epicardium. Following regional ischemia, the ligature is loosened, and the heart is allowed to re-perfuse.
For both survival and non-survival experiments, the myocardial function can be assessed via an echocardiography (ECHO) measurement of the fractional shortening. For non-survival studies, data from sonomicrometry collected using three digital piezoelectric ultrasonic probes implanted within the ischemic area and the left ventricle developed pressure (LVDP) using an apically inserted left ventricle (LV) catheter can be continuously acquired for evaluating the regional and global myocardial function, respectively.
For survival studies, the incision is closed, a left needle thoracentesis is performed for pleural air evacuation, and postoperative pain control is achieved.

The present study demonstrates a highly reproducible animal model of acute regional myocardial ischemia and reperfusion injury in rabbits using a left mini-thoracotomy for survival cases or a midline sternotomy for non-survival cases.

兔子缺血和再灌注损伤模型

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and ...

<meta charset="utf8"></head><body>基于气管切开术的气管插管方案，用于非存活大鼠模型

Endotracheal Intubation via Tracheotomy and Subsequent Thoracotomy in Rats for Non-Survival Applications

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that require controlled or high ventilation pressures or access to the thoracic cavity and organs. Conventional endoorotracheal intubation using the anatomically existing route through the mouth is well suited for survival experiments. However, this procedure poses some challenges, including generally higher levels of the required experience and technical skill, more advanced equipment, and greater time effort with relevant intubation failure rates and complications such as tracheal perforation, temporary systemic hypooxygenation, and relevant aerial leakage.
This manuscript, therefore, presents a detailed step-by-step protocol for endotracheal intubation through tracheotomy in non-survival rat models when guaranteed intubation success, constant oxygenation levels, high ventilation pressures, or open thoracotomy are required.
The protocol emphasizes the importance of meticulous surgical technique to ensure consistent and reliable outcomes, especially for researchers who are inexperienced or lack routine in the technique of endoorotracheal intubation via direct laryngoscopy. This procedure is, therefore, expected to minimize animal suffering and unnecessary animal losses.

<meta charset="utf8"></head><body>大鼠气管切开术气管插管和随后的开胸术用于非存活应用

Here we introduce a standardized procedure for endotracheal intubation via tracheotomy followed by thoracotomy in rats, aimed at enhancing the precision and reproducibility of non-survival applications requiring invasive ventilation and exposure of thoracic organs in in vivo rat models.

大鼠气管切开术气管插管和随后的开胸术用于非存活应用

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that ...

用聚丙烯导管引导家兔气管插管

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新生儿兔子的盲内窥管插管

简化新生早产儿经皮气管内药物输送

简化新生早产儿经皮气管内药物输送

兔模型中的孤立肺灌注系统

使用柔性插管内窥镜作为猪安全气道管理的标准化模型的气管插管

兔子缺血和再灌注损伤模型

大鼠气管切开术气管插管和随后的开胸术用于非存活应用

大鼠气管切开术气管插管和随后的开胸术用于非存活应用