这项研究是通过主动脉研究基金（密歇根大学）给Eliason博士做出的。作者还要感谢密歇根大学工厂运营标志和图形部的Nick Scott，协助设计和组装卷烟照明设备。

以下协议是在密歇根大学体育动物护理和使用委员会（IACUC）的指导和批准下进行的。 电子烟气室组装注意:使用过程中，整个室应放置在通风橱中。这里的室被安置在温度控制和过滤的实验室环境中。调查员可以选择监测系统的这些方面，以确保房间空气质量的一致性。作为选择，用金属笼盖覆盖显示器可以防止啮齿动物篡改，同时允许监视器对室内环境进行采样。 <ol><li>获得体积为20升的带气密性可拆卸盖的麻醉室。 </li><li>使用装有适于切割材料的刀片的夹具锯，在室的盖子上切下10.2厘米直径的孔，距离后边缘大约7.6厘米室。 </li><li>将可调节排气孔插入孔中，并用任何填缝胶粘固定到位。 注意:确保胶囊不能接触到室内的啮齿动物，因为它们可能会堵塞填隙物。为了防止这种可能的问题，将填缝物施加到室壁的外部以安装通风口。 </li><li>将硅胶管切成两个15厘米的段，并将端部连接到T型连接器的两侧。 注意:硅胶管有可能与电子蒸气或香烟烟雾的某些组分发生反应。因此，研究者可以考虑使用非反应性管。 </li><li>通过在室盖前部附近的预制孔将两个硅胶管穿过，使得T型连接器在室内。确保使用粘合剂填缝或电气胶带将管道固定到盖子上。 </li><li>将硅胶管的自由端连接到两个微型气泵的输出端。泵应安装在室内的盖子上，双面胶带或填缝料。 注意:连接到泵的输出端的管子的长度应该很短，以限制使用过程中管道内表面的蒸气收集量。 </li><li>使用新的硅胶管，将一端连接到其中一个空气泵（ 图1中的泵A）的输入侧，并将该管切割成约4厘米长。这是在室使用期间将插入电子烟的地方。确保管的直径允许在电子烟囱末端附近贴合。 </li><li>使用新的硅胶管，将一端连接到另一个空气泵的输入侧（ 图1中的泵B）。该泵将室内空气引入。因此，管的端部必须放置在通风橱的外面。该管的长度不是关键的，但应尽可能短以限制气流阻力。 </li><li>在室内安装两个小钩，双面胶，以保持oxygen和一氧化碳气体监测仪。 </li></ol><img alt="图1" src="/files/ftp_upload/55672/55672fig1.jpg" /> 电子烟雾室示意图。 室内装有通风橱（未显示）。室内空气泵（B泵）将通风柜外部的室内空气以2L / min连续引入室内。 E-cig泵（泵A）在4秒内抽出133毫升的电子蒸气，30秒的休息间隔。电蒸汽和室内空气混合，然后泵入室内。气体监测器连续测量室内的一氧化碳（CO）和氧（O 2 ）浓度。电子蒸气被动地通过通风橱通风。 <a href="https://www.jove.com/files/ftp_upload/55672/55672fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 香烟烟雾室装配注:几乎任何品牌of香烟可以与该系统一起使用，但是标准化的研究香烟，如肯塔基大学1R6F研究香烟是经济高效，可靠和最适合这种应用。 <ol><li>按照步骤1.1 - 1.6和步骤1.9。 </li><li>使用新的硅胶管，将一端连接到香烟照明装置，另一端连接到空气泵的输入侧（ 图2中的泵A）。香烟照明装置应在使用过程中放置​​在通风橱内和室外。 注意:构建香烟照明装置需要知道金属制造和电气工程。虽然这里不再提供一步一步的施工指南，但请参阅计划的补充材料。 </li><li>使用新的硅胶管，将一端连接到另一个空气泵（ 图2中的泵B）。该泵将室内引入室内空气，因此管的端部必须放置在通风橱的外部。 </li<html> &gt; <li>在腔室的前壁中切割几个5 mm宽的垂直狭缝，并将计算机风扇安装在腔室的外部，使其覆盖该开口。确保风扇的前部面向室，使风扇通过该开口将空气吹入室内。 </li></ol><img alt="图2" src="/files/ftp_upload/55672/55672fig2.jpg" /> 图2.香烟烟雾室示意图。 室内空气泵（B泵）将通风柜外部的室内空气以2L / min连续引入室内。泵A以2升/分钟的速度吸入点燃的香烟40秒，20秒后，电脑风扇在3分钟内排空。抽烟和室内空气混合，然后泵入室内。气体监测器连续测量一氧化碳（CO）和氧（O 2 ）室浓度。烟雾通过通风橱排空。ftp_upload / 55672 / 55672fig2large.jpg"target ="_ blank"&gt;请点击此处查看此图的较大版本。 3.微控制器装配和软件<ol><li>通过单独的微控制器控制电子烟雾室和香烟烟雾室的泵送系统。下载微控制器软件并上传S 辅助材料中提供的操作代码。电子香烟蒸汽代码将连续运行室内空气泵，并每30秒激活电子烟泵持续4秒。香烟烟雾代码将连续运行室内空气泵，启动香烟泵40秒，并在香烟泵停止后启动计算机风扇20s。运行3分钟后，风扇将关闭。 注意:泵和风扇的时间可以根据需要进行调整。参考制造商有关如何将相关代码上传到微控制器的说明。 </li><li>装配如图3所示，将拉线，二极管，电阻和电容器连接到面包板，并将鳄鱼夹线连接到相应的空气泵（以及用于香烟烟雾室的计算机风扇）。如果可能的话，将微控制器放在通风橱外面。 </li></ol><img alt="图3" src="/files/ftp_upload/55672/55672fig3.jpg" /> 图3.微控制器示意图。 单片机和面包板的原理图，用于操作空气泵和风扇的时序。 <a href="https://www.jove.com/files/ftp_upload/55672/55672fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 动物<ol><li>使用体重450-520克的成年老鼠。 </li><li>根据暴露类型（ 例如 ，电子烟，香烟烟雾，室内空气泵入室）组大鼠。 </li><li>在这个时候若要暴露，请使用注射器从尾静脉采集EDTA涂层管中的500μL血液，以测量基线血清可替宁浓度。 </li><li>在4℃下以20,000xg的速度旋转30分钟，收集血清。确保样品在整个过程中在冰上冷藏。 </li><li>按照制造商方案，收集血清样品中可替宁的检测。样品也可以储存在-80℃以供以后使用。 </li></ol> 5.操作电子烟雾室<ol><li>用70％乙醇清洗室内，然后用去离子水清洗室内空气，直到室完全干燥（或约30分钟）。 </li><li>校准气体监测器并将整个显示器安装在腔室内的墙壁上。 </li><li>将大鼠放在室内。请注意，最多可以同时暴露三只动物。 </li><li>确保电子烟充满充足的电子液体并插入电子烟进入输入管。在90分钟暴露的持续时间内，确保电子烟电池和电子液体含量是足够的。 </li><li>打开空气泵，然后启动定时器。 </li><li>在曝光期间观察气体监测器以确保室含有&gt; 20％O 2和0ppm CO。 注意:如果氧气含量下降，腔室通风不良或氧气监测仪可能无法适当校准。 </li><li>曝光时间达到90分钟后，取出电子烟，继续运行气泵，排出剩余的蒸气。此外，可以提升室的顶部以加速通风。 </li><li>蒸汽清除后，取出大鼠并清洗腔。 </li><li>在实验方案结束后约1小时，从每只大鼠尾静脉采集500μL血液。 </li><li>按照步骤4.4 - 4.5分离血清和检测可替宁。 </li></ol><ol><li>按照步骤5.1 - 5.3。 </li><li>将香烟插入香烟照明装置，香烟的末端与加热元件相连。 </li><li>打开香烟照明设备，直到烟头开始闷热（约5秒钟）。 </li><li>一旦香烟点亮，打开泵送系统，启动计时器，并观察香烟燃烧完成（约40秒）。 </li><li>一旦香烟燃烧，请用镊子小心地从香烟照明设备中取出废香烟。 </li><li>确保CO含量不超过1000ppm，O 2水平不低于20％。计算机风扇的时序和持续时间对于防止一氧化碳的积聚至关重要。 </li><li> 4分钟后，关闭泵送系统并返回步骤6.2，直到大鼠暴露于烟草烟雾90分钟（或约23支香烟）。 注意:一氧化碳含量在香烟头4分钟后ning应低于400 ppm，否则一氧化碳可能开始积聚在室内。 </li><li>当曝光完成时，继续离开泵送系统以使残余烟雾通风。当一氧化碳低于100ppm时，从室中除去大鼠。这应该需要5 - 10分钟。 </li><li>取出老鼠并清理腔室。 </li><li>在实验方案结束后，大约1小时后从尾静脉收集500μL血液，与电子香烟一样。 </li><li>按照步骤4.4 - 4.5分离血清和检测可替宁。 </li></ol>

<ol>
	<li>Schoenborn, C. A., Gindi, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+Cigarette+Use+Among+Adults:+United+States,+2014+Key+findings.">Electronic Cigarette Use Among Adults: United States, 2014 Key findings.</a> NCHS. , (2014).</li><li>Singh, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tobacco+Use+Among+Middle+and+High+School+Students+-+United+States,+2011-2015.">Tobacco Use Among Middle and High School Students - United States, 2011-2015.</a> MMWR. 65 (14), 361-367 (2016).</li><li>Flora, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+potential+impurities+and+degradation+products+in+electronic+cigarette+formulations+and+aerosols.">Characterization of potential impurities and degradation products in electronic cigarette formulations and aerosols.</a> Regul. Toxicol. Pharmacol: RTP. 74, 1-11 (2016).</li><li>Tierney, P. A., Karpinski, C. D., Brown, J. E., Luo, W., Pankow, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavour+chemicals+in+electronic+cigarette+fluids.">Flavour chemicals in electronic cigarette fluids.</a> Tob. Control. , (2015).</li><li>Sleiman, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emissions+from+Electronic+Cigarettes:+Key+Parameters+Affecting+the+Release+of+Harmful+Chemicals.">Emissions from Electronic Cigarettes: Key Parameters Affecting the Release of Harmful Chemicals.</a> Environ. Sci. Technol. 50 (17), 9644-9651 (2016).</li><li>Hwang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+cigarette+inhalation+alters+innate+immunity+and+airway+cytokines+while+increasing+the+virulence+of+colonizing+bacteria.">Electronic cigarette inhalation alters innate immunity and airway cytokines while increasing the virulence of colonizing bacteria.</a> Int J Mol Med (Berlin, Germany). 94 (6), 667-679 (2016).</li><li>Cheng, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+evaluation+of+electronic+cigarettes.">Chemical evaluation of electronic cigarettes.</a> Tob Control. 23, 11-18 (2014).</li><li>Etter, J. -. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+longitudinal+study+of+cotinine+in+long-term+daily+users+of+e-cigarettes.">A longitudinal study of cotinine in long-term daily users of e-cigarettes.</a> Drug Alcohol Depend. 160, 218-221 (2016).</li><li>Etter, J. -. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Levels+of+saliva+cotinine+in+electronic+cigarette+users.">Levels of saliva cotinine in electronic cigarette users.</a> Addiction. 109 (5), 825-829 (2014).</li><li>Marsot, A., Simon, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nicotine+and+Cotinine+Levels+With+Electronic+Cigarette:+A+Review.">Nicotine and Cotinine Levels With Electronic Cigarette: A Review.</a> Int. J. Toxicol. 35 (2), 179-185 (2016).</li><li>Flouris, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+impact+of+active+and+passive+electronic+cigarette+smoking+on+serum+cotinine+and+lung+function.">Acute impact of active and passive electronic cigarette smoking on serum cotinine and lung function.</a> Inhal. Toxicol. 25 (2), 91-101 (2013).</li><li>Bot, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+cotinine+levels+in+cigarette+smokers:+impact+of+mental+health+and+other+correlates.">Plasma cotinine levels in cigarette smokers: impact of mental health and other correlates.</a> Eur Addict Res. 20 (4), 183-191 (2014).</li><li>Zhu, S. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Four+hundred+and+sixty+brands+of+e-cigarettes+and+counting:+implications+for+product+regulation.">Four hundred and sixty brands of e-cigarettes and counting: implications for product regulation.</a> Tob. Control. 23, 3-9 (2014).</li></ol>

作者没有什么可以披露的。

在这里，我们描述了一种建立以受控方式将啮齿动物暴露于电子烟和香烟烟雾的室的方法（ 图6 ）。与商业曝光系统14,15,16相比，电子烟雾室的构造相对简单和便宜。建立房间所需的部件和工具可从商业供应商在线获得。类似地，除了必须制造的香烟照明装置之外，构建香烟烟雾室是相对简单的（参见用于计划的补充材料 ）。 一旦房间被构造，暴露系统的关键步骤就是对室进行校准，以将啮齿动物暴露于希望量的尼古丁。在电子烟盒和香烟盒中，调整总曝光量time也许是增加或减少尼古丁暴露量的最简单的方法。增加电子香烟暴露系统中的抽吸时间可能会增加尼古丁的剂量，但是延长电子烟卷烟的蒸气长时间已被证明会增加ROS，醛类和其他有害化合物的含量，或者可能不反映电子烟用户的典型习惯5 。可以通过修改上传到微控制器的代码来调整抽吸持续时间和总曝光时间。此外，应注意的是，电子烟溶液中的尼古丁浓度以及电子烟加热元件的电压可能会发生显着变化，在校准系统时应予以考虑。 这种曝光系统的最大优点之一是其多功能性。几乎任何品牌的e-cig或e-cig解决方案都可以用于该系统。这是一个特别有用的功能因为电子烟市场现在包括400多个品牌和数以千计的电子烟解决方案13 。此外，暴露系统与多个实验终点兼容，允许电子香烟对各种器官系统和疾病过程的影响进行研究。我们还承认，这种暴露范例有几个限制，例如动物暴露于蒸气的方法。电子烟用户直接吸入电子蒸气，而在这种范式中，啮齿动物被动地吸入电子蒸气。此外，啮齿动物还可能通过其他途径吸收蒸汽或烟雾中的化合物（ 即 ，直接吸收皮肤，并在吸食时）。然而，我们认为曝光系统的好处远远超过了限制。 总的来说，这种曝光范例提供了一致和临床相关的电子烟和香烟烟雾暴露，并可能有所帮助帮助研究工作确定电子烟和香烟烟雾对健康的负面影响。

自从2004年进入美国市场以来，电子香烟（电子香烟）已经扩大到十亿美元的行业，据估计有近900万成年人经常使用它们1 。在2014年和2015年，更多的高中生使用电子香烟比传统香烟2 。越来越多的电子烟用户已经开展了一项研究工作，以评估其潜在的不利健康影响。 电子香烟通过加热通常含有水，聚乙二醇或植物甘油，尼古丁和调味剂3,4的混合物的粘稠溶液产生蒸汽（称为"E蒸气"）。已经表明，电子蒸气含有几种有害化合物，包括活性氧物质（ROS），尼古丁，各种醛和多环芳烃5 ，6。许多这些化合物在吸入之前的E液体的蒸发过程中形成7 。值得注意的是，这些有害化合物中的几种也存在于香烟烟雾中，引起人们关注的是，电子烟使用可能具有类似的不良健康后果7 。 对电子香烟的健康影响几乎没有共识。为了解决这个问题，已经开发了几种电子蒸气暴露的动物模型（ 表1 ）。这些模型采用各种方法，如全身电子蒸气暴露和机械通气。虽然目前的模型已经提供了有见地的数据，但很少能够在同一曝光系统中直接比较香烟烟雾（ 表1 ）。此外，虽然几项人类研究显示，电子烟用户和吸烟者的血清可替宁水平在30-200ng / mL之间，但许多型号的电子蒸气和香烟烟雾暴露量下降这个范围是8，9，10，11，12。 在这里，我们提出一种比较香烟烟雾和体内电子蒸气暴露的影响的方法 ，产生与人类研究相似的血清可替宁水平。

<table border="0" cellpadding="0" cellspacing="0" width="793">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:348px;">blu PLUS Rechargeable Kit</td>
			<td style="width:219px;">blu eCigs</td>
			<td style="width:119px;">N/A</td>
			<td style="width:107px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1R6F Reference Cigarettes</td>
			<td>Center for Tob Ref&#160; Prod UK</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lexan Anesthesia Chamber 20 L</td>
			<td>Jorgensen Laboratories</td>
			<td>JOR265</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Arduino UNO</td>
			<td>Arduino</td>
			<td>2877</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Diode Rectifier - 1A 50V</td>
			<td>Spark Fun</td>
			<td>COM-08589</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Resistor 10K Ohm 1/6th Watt PTH - 20 pack</td>
			<td>Spark Fun</td>
			<td>COM-11508</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Electrolytic Decoupling Capacitors - 100uF/25V</td>
			<td>Spark Fun</td>
			<td>COM-00096</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Solderless Plug-in BreadBoard&#160;</td>
			<td>BusBoard Prototype Systems</td>
			<td>BB400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alligator-Clip Wires</td>
			<td>BusBoard Prototype Systems</td>
			<td>CA-M-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ZipWire</td>
			<td>BusBoard Prototype Systems</td>
			<td>ZW-MM-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard Fan 80 ST2</td>
			<td>Cooler Master</td>
			<td>R4-S8R-20AK-GP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ARIC 4&#34; adjustable vent</td>
			<td>Bestlouver</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ToxiPro&#160; Carbon Monoxide Monitor</td>
			<td>Honeywell Analytics</td>
			<td>54-00-10316</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ToxiPro Oxygen Monitor</td>
			<td>Honeywell Analytics</td>
			<td>54-45-90-VD</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ToxiPro IQ Express Docking Station</td>
			<td>Honeywell/Sperian Biosystems&#160;</td>
			<td>54-46-9100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Command Wall Hook Small Wire 6-Pack</td>
			<td>3M</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro Water/Air Pump</td>
			<td>Xiamen Conjoin Electronics</td>
			<td>CJWP40-A12A1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1/4&#34; Silicon Tubing</td>
			<td>NewAge</td>
			<td>2801470-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">T Connector</td>
			<td>Bel-Art Scienceware</td>
			<td>F196060000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plastic Whole Blood tube with spray-coated K2EDTA</td>
			<td>Becton, Dickinson and Company</td>
			<td>367841</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotinine ELISA kit</td>
			<td>Calbiotech</td>
			<td>CO096D</td>
			<td></td>
		</tr>
	</tbody>
</table>

comparing the effects of electronic cigarette vapor and cigarette smoke in a novel in vivo exposure system

<html>电子香烟（电子烟 ）正在被广泛使用，并越来越受欢迎。据估计有超过900万成年人经常使用它们。电子烟雾（E-vapor）暴露潜在的不良健康影响定义不明确。虽然已经开发了几种动物模型的电子 蒸气暴露，但很少有模型将啮齿动物暴露于临床相关量的尼古丁，并在同一暴露系统内直接与香烟烟雾进行比较。在这里，我们提出了一个构建和操作E 蒸气室和香烟烟雾室的方法。这些室是通过装备有麻醉室的计算机控制的泵送系统构成的，其输送一定量的E尼古丁暴露量通过量化前后暴露后的血清可替宁水平进行间接测量，该暴露系统可以修改以适应各种类型的电子 烟和烟草香烟，并且可以用于比较电子 烟和香烟烟雾在体内的影响 。

一氧化碳和氧气监测在电子蒸气暴露过程中，氧浓度不低于20％，CO浓度在整个曝光期间仍然不可检测。香烟烟雾暴露期间的气体监测仪表明，氧气浓度保持在20％以上。一氧化碳浓度不超过1,000ppm（ 图4 ）。 <img alt="图4" src="/files/ftp_upload/55672/55672fig4.jpg" /> 图4:香烟烟雾室中的一氧化碳浓度。在引入来自1R6F香烟的烟雾的过程中，每30秒记录一氧化碳浓度。显示的结果是3连续4分钟循环的平均值。一氧化碳浓度不超过1,000ppm。香烟在40秒内燃烧完成，风扇启动d 20秒后（ 即点烟后1分钟风扇启动）。 前和后暴露血清Cotinine E-vapor组（n = 3）的暴露前和1小时暴露后血清可替宁分别为4.2±0.4ng / mL和171.6±20.5ng / mL。吸烟前后1小时吸烟组（n = 3）后血清可替宁分别为3.9±0.3 ng / mL和98.8±2.1 ng / mL（ 图5 ）。 <img alt="图5" src="/files/ftp_upload/55672/55672fig5.jpg" /> 图5:香烟烟雾或电子烟雾暴露后的血清Cotinine水平。血清可替宁在方案开始前和在90分钟暴露时间后1小时测量。电子蒸汽组暴露前和暴露后1小时血清可替宁为4.2±0.4 ng / mL and为171.6±20.5ng / mL。吸烟前1小时和吸烟后1小时血清可替宁分别为3.9±0.3 ng / mL和98.8±2.1 ng / mL。前后暴露后血清可替宁浓度差异具有统计学意义。 * P &lt;0.05。 <img alt="图6" src="/files/ftp_upload/55672/55672fig6.jpg" /> 图6:电子烟雾室和香烟烟雾室。通风橱内电子烟蒸气（右）和香烟烟雾室（左）的图像。红色框包含微控制器。 <a href="https://www.jove.com/files/ftp_upload/55672/55672fig6large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td colspan="2"> 动物模型 </td><td colspan="2"> &lt;强&gt;电子烟</td><td colspan="2"> 烟草卷烟 </td><td> 参考 </td></tr><tr><td> 曝光方法 </td><td> 模型生物 </td><td> 品牌（尼古丁） </td><td> 氨苄青霉素/ mL（血清，[尿]） </td><td> 牌 </td><td> 氨苄青霉素/ mL（血清，[尿]） </td><td></td></tr><tr><td>全身暴露</td><td> C57BL / 6J小鼠</td><td> Joytech 510-T（1.8％） </td><td> 62.3±3.3，[892.5±234] </td><td> N / A </td><td> N / A </td><td>麦格拉思 - 莫罗</td></tr><tr><td>机械呼吸机</td><td> BALB / cJ小鼠</td><td>没有报道</td><td> [400 - 500] </td><td>没有报道</td><td> [500 - 800] </td><td> Ponzoni </td></tr><tr><td>全身暴露</td><td> CD-1小鼠</td><td>多重*（0.6 - 24％） </td><td>没有报道</td><td> N /一个</td><td> N / A </td><td>黄禹锡</td></tr><tr><td>全身暴露</td><td> Wistar白化大鼠</td><td>自由T（0.9％） </td><td>没有报道</td><td>没有报道</td><td>没有报道</td><td> Salturk </td></tr><tr><td>全身暴露</td><td> C57BL / 6小鼠</td><td> NJOY（1.8％） </td><td> 267±17 </td><td> N / A </td><td> N / A </td><td> Sussan </td></tr><tr><td>全身暴露</td><td> C57BL / 6J小鼠</td><td> CoolCart，蒸汽泰坦</td><td> 500±10 </td><td> 3R4F参考香烟</td><td> 76±7.6 </td><td> Husari </td></tr><tr><td colspan="5"> * Xtreme Vaping，Vapure，Vape Addict Juice，Grimm Creations，Green Smart Living，Free Masons Elixer </td><td></td><td></td></tr></tbody></table> 表1:电子烟雾暴露模型的特征。

Watch this Scientific Journal Video about Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System at JoVE.com

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

该协议描述了将啮齿动物暴露于电子烟蒸气（E蒸汽）和香烟烟雾的方法。通过用自动泵送系统修改麻醉室来构建暴露室，其将电子蒸气或香烟烟雾传递给啮齿动物。该系统可以轻松修改，以适应许多实验端点。

比较电子烟和烟雾在小说中的影响<em&gt;体内</em&gt;曝光系统

Electronic cigarettes (E-cigarettes) are being widely used, and growing in popularity. It is estimated that more than 9 million adults use them regularly. ...

comparing-effects-electronic-cigarette-vapor-cigarette-smoke-novel

Universidad San Sebastian

Research

JoVE Journal

Medicine

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

11.4K 次观看.  University of Michigan Medical School. 该协议描述了将啮齿动物暴露于电子烟蒸气（E蒸汽）和香烟烟雾的方法。通过用自动泵送系统修改麻醉室来构建暴露室，其将电子蒸气或香烟烟雾传递给啮齿动物。该系统可以轻松修改，以适应许多实验端点。 

视频: Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

Making Sense of Listening: The IMAP Test Battery

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.

A test battery (IMAP) for performing an in-depth assessment of auditory and cognitive abilities contributing to listening skills is described. It is quick to administer, child-friendly and free from linguistic confounds. Stimulus generation and protocol management are controlled via a software platform (IHR-STAR) to ensure replicable procedures.

听力:IMAP测试电池的决策意识

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the ...

科研

医学

Isolation of Mouse Respiratory Epithelial Cells and Exposure to Experimental Cigarette Smoke at Air Liquid Interface

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated respiratory epithelium. A protocol is described for isolating and exposing these cells to mainstream cigarette smoke (CS), in order to study epithelial cell responses to CS exposure. The protocol consists of three parts: the isolation of airway epithelial cells from mouse trachea, the culturing of these cells at air-liquid interface (ALI) as fully differentiated epithelial cells, and the delivery of calibrated mainstream CS to these cells in culture. The ALI culture system allows the culture of respiratory epithelia under conditions that more closely resemble their physiological setting than ordinary liquid culture systems. The study of molecular and lung cellular responses to CS exposure is a critical component of understanding the impact of environmental air pollution on human health. Research findings in this area may ultimately contribute towards understanding the etiology of chronic obstructive pulmonary disease (COPD), and other tobacco-related diseases, which represent major global health problems.

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface as a model of differentiated respiratory epithelium. A protocol is described for isolating, culturing and exposing these cells to mainstream cigarette smoke, in order to study molecular responses to this environmental toxin.

小鼠呼吸道上皮细胞和实验香烟烟雾暴露在空气分离液界面

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated ...

A Strategy to Identify de Novo Mutations in Common Disorders such as Autism and Schizophrenia

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, the de novo mutation rate in humans is relatively high, so new mutations are generated at a high frequency in the population. However, de novo mutations have not been reported in most common diseases. Mutations in genes leading to severe diseases where there is a strong negative selection against the phenotype, such as lethality in embryonic stages or reduced reproductive fitness, will not be transmitted to multiple family members, and therefore will not be detected by linkage gene mapping or association studies. The observation of very high concordance in monozygotic twins and very low concordance in dizygotic twins also strongly supports the hypothesis that a significant fraction of cases may result from new mutations. Such is the case for diseases such as autism and schizophrenia. Second, despite reduced reproductive fitness1 and extremely variable environmental factors, the incidence of some diseases is maintained worldwide at a relatively high and constant rate. This is the case for autism and schizophrenia, with an incidence of approximately 1% worldwide. Mutational load can be thought of as a balance between selection for or against a deleterious mutation and its production by de novo mutation. Lower rates of reproduction constitute a negative selection factor that should reduce the number of mutant alleles in the population, ultimately leading to decreased disease prevalence. These selective pressures tend to be of different intensity in different environments. Nonetheless, these severe mental disorders have been maintained at a constant relatively high prevalence in the worldwide population across a wide range of cultures and countries despite a strong negative selection against them2. This is not what one would predict in diseases with reduced reproductive fitness, unless there was a high new mutation rate. Finally, the effects of paternal age: there is a significantly increased risk of the disease with increasing paternal age, which could result from the age related increase in paternal de novo mutations. This is the case for autism and schizophrenia3. The male-to-female ratio of mutation rate is estimated at about 4&#x2013;6:1, presumably due to a higher number of germ-cell divisions with age in males. Therefore, one would predict that de novo mutations would more frequently come from males, particularly older males4. A high rate of new mutations may in part explain why genetic studies have so far failed to identify many genes predisposing to complexes diseases genes, such as autism and schizophrenia, and why diseases have been identified for a mere 3% of genes in the human genome. Identification for de novo mutations as a cause of a disease requires a targeted molecular approach, which includes studying parents and affected subjects. The process for determining if the genetic basis of a disease may result in part from de novo mutations and the molecular approach to establish this link will be illustrated, using autism and schizophrenia as examples.

Molecular genetic strategy for finding de novo mutations causing common disorders such as autism and schizophrenia.

一项战略，以确定在常见的疾病，如自闭症和精神分裂症的新突变

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, ...

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

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 Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

一种新型的<em&gt;在体内</em&gt;基因转移技术和<em&gt;在体外</em&gt;细胞分析为肌肉骨骼疾病骨质流失的研究

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

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

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

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

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

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

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

COPD is projected to be the third most common cause of mortality world-wide by 2020(1). Animal models of COPD are used to identify molecules that contribute to the disease process and to test the efficacy of novel therapies for COPD. Researchers use a number of models of COPD employing different species including rodents, guinea-pigs, rabbits, and dogs(2). However, the most widely-used model is that in which mice are exposed to cigarette smoke. Mice are an especially useful species in which to model COPD because their genome can readily be manipulated to generate animals that are either deficient in, or over-express individual proteins. Studies of gene-targeted mice that have been exposed to cigarette smoke have provided valuable information about the contributions of individual molecules to different lung pathologies in COPD(3-5). Most studies have focused on pathways involved in emphysema development which contributes to the airflow obstruction that is characteristic of COPD. However, small airway fibrosis also contributes significantly to airflow obstruction in human COPD patients(6), but much less is known about the pathogenesis of this lesion in smoke-exposed animals. To address this knowledge gap, this protocol quantifies both emphysema development and small airway fibrosis in smoke-exposed mice. This protocol exposes mice to CS using a whole-body exposure technique, then measures respiratory mechanics in the mice, inflates the lungs of mice to a standard pressure, and fixes the lungs in formalin. The researcher then stains the lung sections with either Gill&#8217;s stain to measure the mean alveolar chord length (as a readout of emphysema severity) or Masson&#8217;s trichrome stain to measure deposition of extracellular matrix (ECM) proteins around small airways (as a readout of small airway fibrosis). Studies of the effects of molecular pathways on both of these lung pathologies will lead to a better understanding of the pathogenesis of COPD.

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

肺气肿，小气道重塑的香烟烟雾暴露小鼠自动测量

COPD is projected to be the third most common cause of mortality world-wide by 2020(1).  Animal models of COPD are used to identify molecules that ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...