我们感谢珀斯儿童医院基金会 （Perth Children's Hospital Foundation） 在购买本项目中使用的脑电图设备方面的慷慨支持。我们感谢 Gardar Thorvardsson （Kvikna） 和 Ieesha Sparks （Temple Medical and Scientific） 在研究项目的技术方面的支持。

该研究获得了儿童和青少年健康服务 （CAHS） 人类研究伦理委员会（HREC，批准号 RGS0000004988）的批准，并遵守了赫尔辛基宣言的原则。
1. 确定纳入研究的患者
<ol><li>在转诊时确定婴儿是否有资格参加研究。任何疑似 HIE 需要运输的婴儿都有资格参加该研究。</li>
	<li>使用标准化诊断标准12 识别疑似 HIE 婴儿：<ol><li>检查胎龄是否为 >35 周。</li>
		<li>检查是否在出生后 <6 小时。</li>
		<li>检查是否存在窒息的证据，如存在以下 4 项标准：<ol><li>检查是否有任何可能导致 HIE 的急性围产期事件（例如，胎盘早剥、脐带脱垂、严重的 FHR 异常）。</li>
			<li>检查 10 分钟时 Apgar 评分是否为 <6，或者在 10 分钟时是否需要继续进行正压通气 +/- 胸外按压复苏。</li>
			<li>出生后 60 分钟内检查脐带 pH 值是否为 <7.0 或 BE 是否为 >12 mmol/L（如果能够进行气体呼吸）。</li>
			<li>如果脐带 pH 值不可用，请在出生后 60 分钟内检查动脉 pH 值是否为 <7.0 或 BE 是否为 >12 mmol/L（如果能够进行气体呼吸）。</li>
		</ol></li>
	</ol></li>
</ol>2. 获得父母的同意
<ol><li>到达后，与父母联系，以获得使用该设备的口头同意，并对其使用的可接受性进行调查。</li>
	<li>提供父母信息表，其中包含知情同意所需的所有信息。</li>
	<li>确保家长理解信息并有机会提出任何问题。</li>
	<li>在口头同意后 72 小时内获得书面同意。</li>
</ol>3. 请求运输医生和护士的同意
<ol><li>请求运输医生和护士同意向他们发送有关他们在运输中使用 aEEG 的经验的调查。</li>
</ol>4. aEEG 应用
<ol><li>按照标准指南13 将 aEEG 电极针插入婴儿的头皮。</li>
	<li>测量并定位解剖标志：矢状缝和耳屏。</li>
	<li>将电极卷尺（图 1）垂直对准头部并平行于面部。</li>
	<li>匹配定位辅助工具上的字母/符号，直到两个位置出现相同的字母/符号（图 1）。</li>
	<li>用手术笔在胶带的每一侧标记位置（图 2）。<ol><li>让头发自然部分，或使用无菌水和棉签棒将其垂直分开，远离标记位置。用纱布擦干。</li>
		<li>用 1% 洗必泰/酒精棉签清洁皮肤并晾干。</li>
		<li>紧紧握住皮肤，以 30° 角插入针头，传感器线朝上。</li>
		<li>使用消毒条和 Chevron 方法14 将皮下针电极固定到位（图 3）。</li>
		<li>对其他针插入重复前面的步骤。</li>
		<li>如果头皮毛茸茸，请在针头插入部位周围使用皮肤准备棉签，并在贴布前晾干。</li>
		<li>将针和电线指向同一方向。</li>
		<li>将参比电极放在胸部（前部或后部）、肩部后部或颈部的后颈区域。</li>
	</ol></li>
	<li>使用 C3/C4 和 P3/P4 配置将电极连接到 aEEG 放大器（参见 图 4）。</li>
</ol>5. 设置脑电图放大器和平板电脑
<ol><li>连接 amplifier 使用以太网电缆连接到平板电脑; 图 5 中显示了连接的概述。</li>
	<li>打开平板电脑并登录。</li>
	<li>进入 Acquire Pro 应用程序。将自动应用默认的新生儿配置。</li>
	<li>将阻抗的默认设置从 5 KΩ 更改为 10 KΩ。</li>
	<li>按 Start Recording（开始记录），将自动创建患者 ID。</li>
	<li>按 EEG 选项卡观察实时轨迹。</li>
	<li>将临床片剂固定在婴儿床上。在 Mansell 新婴儿床或 Voyager 运输培养箱中运输婴儿（图 6 和 图 7）。平板电脑在婴儿床上的位置取决于运输时带走的婴儿床（见 图 8）。</li>
	<li>如果使用飞机，请从婴儿床上取下平板电脑并将其握在手中。</li>
</ol>6. 运输过程中的 aEEG 监测。
<ol><li>在运输过程中，在平板电脑软件上记下任何可能导致伪影、临床变化或癫痫发作的动作。使用平板电脑上可按下的按钮来标记重要事件，包括癫痫发作、患者护理、装载/卸载、起飞和着陆。</li>
	<li>对于癫痫发作等连续临床事件，将为其发生的持续时间添加标签。活动结束后，再次按下按钮。对于非连续事件，只需按一次时间戳按钮。</li>
	<li>到达接收医院后，将婴儿转移到重症监护病床。断开头皮电极与研究放大器的连接，并将其转移到 NICU aEEG 监视器上，在那里可以按单位协议继续记录。 注意：治疗临床医生必须对 aEEG 轨迹不知情，并且不得根据轨迹做出紧急决定。</li>
</ol>7. 数据解释
<ol><li>除了脑电图外，还记录人口统计和交通信息。</li>
	<li>使用 Centrum 应用程序下载 aEEG 跟踪。</li>
	<li>确保 aEEG 轨迹由 CPI 和两名具有 5-20 年新生儿 aEEG 读取经验的独立评估员审查。</li>
	<li>使评估员对临床信息和跟踪不知情。</li>
	<li>将跟踪分成 15 分钟的分段，并查看构件分段的数量。如果三位“专家”中的两位同意，则跟踪的一段被视为伪影。已选择任意值 100 μV，此时表示发生移动伪影。</li>
	<li>以 8 Hz 的速率从 C3-C4 电极下载每条迹线的原始数据。 注意：阻抗 （Z） 是电流的电阻，测量皮肤电极放置和导联运动伪影的质量。低、相等和稳定的阻抗可确保最佳 aEEG。软件系统将阻抗记录为设置为 >10 KΩ 的离散通知。记录每条轨迹的量级和数量。</li>
	<li>在运输后使用电子数据采集平板电脑应用程序完成员工和家长调查。 注意：该研究旨在招募和收集 20 名疑似 HIE 患者的数据。</li>
</ol>8. 分析
<ol><li>使用中位数（四分位距;IQR）。</li>
	<li>将对象显示为总跟踪的百分比。将运动伪影描述为平均值（标准差;SD） 总迹线的百分比。</li>
	<li>将 Z 表示为平均值 （范围），将 Z 幅度表示为中位数 （IQR） 表示千欧姆 （KΩ）。</li>
	<li>将跟踪定义为 80% 的时间内 Z 的总计和无移动伪影。 注：该方法中呈现的代表性结果表示为中位数和四分位数间距以及使用 Mann-Whitney U 检验的中位数比较。</li>
</ol>

使用便携式振幅积分脑电图进行婴儿运输

<ol>
	<li>Kurinczuk, J. J., White-Koning, M., Badawi, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+neonatal+encephalopathy+and+hypoxic-ischaemic+encephalopathy.">Epidemiology of neonatal encephalopathy and hypoxic-ischaemic encephalopathy.</a> Early Hum Dev. 86 (6), 329-338 (2010).</li><li>. Midwives Notification System, Government of Western Australia, Department of Health Available from: <a target="_blank" href="https://www.health.wa.gov.au/Reports-and-publications/Western-Australias-Mothers-and-Babies-summary-information/data?report=mns_birth_y">https://www.health.wa.gov.au/Reports-and-publications/Western-Australias-Mothers-and-Babies-summary-information/data?report=mns_birth_y</a> (2023)</li><li>Shankaran, 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=Whole-body+hypothermia+for+neonates+with+hypoxic-ischemic+encephalopathy.">Whole-body hypothermia for neonates with hypoxic-ischemic encephalopathy.</a> N Engl J Med. 353 (15), 1574-1584 (2005).</li><li>Thoresen, 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=Time+is+brain:+Starting+therapeutic+hypothermia+within+three+hours+after+birth+improves+motor+outcome+in+asphyxiated+newborns.">Time is brain: Starting therapeutic hypothermia within three hours after birth improves motor outcome in asphyxiated newborns.</a> Neonatology. 104 (3), 228-233 (2013).</li><li>Wall&#248;e, L., Hjort, N. L., Thoresen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Start+cooling+as+soon+as+possible.">Start cooling as soon as possible.</a> Acta Paediatr. 108 (4), 771 (2019).</li><li>Skranes, J. H., 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=Amplitude-integrated+electroencephalography+improves+the+identification+of+infants+with+encephalopathy+for+therapeutic+hypothermia+and+predicts+neurodevelopmental+outcomes+at+2+years+of+age.">Amplitude-integrated electroencephalography improves the identification of infants with encephalopathy for therapeutic hypothermia and predicts neurodevelopmental outcomes at 2 years of age.</a> J Pediatr. 187, 34-42 (2017).</li><li>Shalak, L. F., Laptook, A. R., Velaphi, S. C., Perlman, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amplitude-integrated+electroencephalography+coupled+with+an+early+neurologic+examination+enhances+prediction+of+term+infants+at+risk+for+persistent+encephalopathy.">Amplitude-integrated electroencephalography coupled with an early neurologic examination enhances prediction of term infants at risk for persistent encephalopathy.</a> Pediatrics. 111 (2), 351-357 (2003).</li><li>Hellstrom-Westas, L., Rosen, I., Svenningsen, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Predictive+value+of+early+continuous+amplitude+integrated+eeg+recordings+on+outcome+after+severe+birth+asphyxia+in+full+term+infants.">Predictive value of early continuous amplitude integrated eeg recordings on outcome after severe birth asphyxia in full term infants.</a> Arch Dis Child Fetal Neonatal Ed. 72 (1), F34-F38 (1995).</li><li>Davies, A., Wassink, G., Bennet, L., Gunn, A. J., Davidson, J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+we+further+optimize+therapeutic+hypothermia+for+hypoxic-ischemic+encephalopathy.">Can we further optimize therapeutic hypothermia for hypoxic-ischemic encephalopathy.</a> Neural Regen Res. 14 (10), 1678 (2019).</li><li>Davis, J. W., 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=Outcomes+to+5+years+of+outborn+versus+inborn+infants+&amp;lt;32+weeks+in+western+australia:+A+cohort+study+of+infants+born+between+2005+and+2018.">Outcomes to 5 years of outborn versus inborn infants &amp;lt;32 weeks in western australia: A cohort study of infants born between 2005 and 2018.</a> Arch Dis Child Fetal Neonatal Ed. 108 (5), 499-504 (2023).</li><li>Schomer, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ambulatory+EEG+monitoring,+reviewing,+and+interpreting.">Ambulatory EEG monitoring, reviewing, and interpreting.</a> J Clin Neurophysiol. 38 (2), 77-86 (2021).</li><li>Hypoxic ischaemic encephalopathy (HIE). Neonatology Available from: <a target="_blank" href="https://cahs.health.wa.gov.au/~/media/HSPs/CAHS/Documents/Health-Professionals/Neonatology-guidelines/Hypoxic-Ischaemic-Encephalopathy.pdf?thn=0">https://cahs.health.wa.gov.au/~/media/HSPs/CAHS/Documents/Health-Professionals/Neonatology-guidelines/Hypoxic-Ischaemic-Encephalopathy.pdf?thn=0</a> (2018)</li><li>Brainz Monitor: Low impedance needle electrodes. Neonatology Available from: <a target="_blank" href="https://cahs.health.wa.gov.au/-/media/HSPs/CAHS/Documents/Health-Professionals/Neonatology-guidelines/Brainz-Monitor-Low-Impedance-Needle-Electrodes.pdf?thn=0">https://cahs.health.wa.gov.au/-/media/HSPs/CAHS/Documents/Health-Professionals/Neonatology-guidelines/Brainz-Monitor-Low-Impedance-Needle-Electrodes.pdf?thn=0</a> (2021)</li><li>Found, P. W. H., Baines, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+securing+cannulae+with+different+taping+methods).">Efficacy of securing cannulae with different taping methods).</a> Anaesth Intensive Care. 28 (5), 547-551 (2000).</li><li>Stephens, C. 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=Electroencephalography+quality+and+application+times+in+a+pediatric+emergency+department+setting:+A+feasibility+study.">Electroencephalography quality and application times in a pediatric emergency department setting: A feasibility study.</a> Pediatr Neurol. 148, 82-85 (2023).</li><li>Variane, G. F. T., 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=Remote+monitoring+for+seizures+during+therapeutic+hypothermia+in+neonates+with+hypoxic-ischemic+encephalopathy.">Remote monitoring for seizures during therapeutic hypothermia in neonates with hypoxic-ischemic encephalopathy.</a> JAMA Netw Open. 6 (11), e2343429 (2023).</li><li>Bruns, N., 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=Application+of+an+amplitude-integrated+EEG+monitor+(cerebral+function+monitor)+to+neonates.">Application of an amplitude-integrated EEG monitor (cerebral function monitor) to neonates.</a> J Vis Exp. (127), e55985 (2017).</li><li>Dilena, R., 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=Consensus+protocol+for+eeg+and+amplitude-integrated+eeg+assessment+and+monitoring+in+neonates.">Consensus protocol for eeg and amplitude-integrated eeg assessment and monitoring in neonates.</a> Clin Neurophysiol. 132 (4), 886-903 (2021).</li><li>Moghadam, S. 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=An+automated+bedside+measure+for+monitoring+neonatal+cortical+activity:+A+supervised+deep+learning-based+electroencephalogram+classifier+with+external+cohort+validation.">An automated bedside measure for monitoring neonatal cortical activity: A supervised deep learning-based electroencephalogram classifier with external cohort validation.</a> Lancet Digit Health. 4 (12), e884-e892 (2022).</li></ol>

作者没有要披露的利益冲突示例。Kvikna ehf.冰岛雷克雅未克和澳大利亚新南威尔士州悉尼的 Temple Medical and Scientific 没有参与研究的设计、分析或解释，但资助了出版费。

这项新颖的研究描述了 aEEG 在出生后不久需要转运的疑似 HIE 婴儿中的应用和早期数据采集。以前没有报道过新生儿运输过程中的 aEEG 应用。aEEG 已在新情况下使用，例如巴西远程医疗神经重症监护咨询和儿科 ED15,16 下的远程监测。
aEEG 针应用类似于“静态”新生儿重症监护17 中使用的针头应用，位于头皮上以实现...

出生时向大脑的氧气输送或血流受损会导致脑损伤（缺氧缺血性脑病;HIE）。这是足月婴儿死亡和残疾的主要原因1。据报道，HIE 的全球发病率为每 1000 次分娩 2 次，中低收入国家的发病率可能更高。在西澳大利亚州，每年大约有 50 名临床诊断为 HIE 的婴儿出生（2022 年有 32308 名活产婴儿，发病率约为 1.5/10002）。
HIE 的诊断标准基于出生情况、婴儿血酸平衡的变化（出生后来自脐带或婴儿）以及神经状态的变化3。中度或重度 HIE 的治疗方法是 33-34 °C 的全身低温。 越早开始，对婴儿神经系统结果的益处就越大 3,4,5。振幅整合脑电图 （aEEG） 是评估和监测的重要组成部分，在许多地区仅在三级中心提供。将电极（EEG 安全 DIN 连接器或带有 1.5 毫米直径引脚的 DIN 42802）插入头皮，以测量和记录大脑电活动。然后将电极连接到放大器，放大器又连接到临床平板电脑以显示和记录脑电波活动。aEEG 可以提供有关脑电波活动的信息，并用于协助诊断 HIE6。神经系统检查和 aEEG 的结合可能会增强识别患有中度或重度 HIE 的婴儿的能力 7,8,9。
西澳大利亚新生儿紧急运输服务 （NETS WA） 每年为大约 1100 名患病婴儿提供重症监护运输服务。当地医院的医护人员可以通过免费电话号码联系 NETS WA，并立即通过电话会议与新生儿专科医生和护士联系以请求转移。NETS 团队乘坐专门的公路救护车或带有专门制造的重症监护运输设备床的固定翼飞机旅行。转运团队到达转诊医院后，对婴儿进行评估，并继续或升级治疗。然后，该团队将婴儿转移到该州首府（珀斯）的三级新生儿重症监护室 （NICU）。在西澳大利亚州，由于转诊医院和集中专科服务10 之间的距离通常很远（最大距离为 1381 英里），aEEG 可能在出生后长达数小时内无法应用，从而导致治疗延误。
许多监测系统是为重症监护病房设计的，但由于其尺寸和功率要求，它们不适用于运输环境。基于病房的设备容易出现移动伪影，这可能会在运输环境中记录的质量数据不足。动态脑电图放大器技术主要为患者在家中佩戴而设计，可以连接到带有脑电图检测软件的计算机平板电脑，并在运输过程中应用于新生儿11。这可以由新生儿转运团队携带给疑似 HIE 的婴儿。该软件能够通过便携式互联网上行链路和基于 Web 的查看器进行远程查看。目前，没有关于 aEEG 在运输环境中的适用性或可读性的报告。
以前没有报道过在转运 HIE 婴儿时应用和监测 aEEG，也没有现有的使用方案。没有证据支持运输环境的可行性或实用性。我们提出了一种在新生儿运输中使用 aEEG 的方案。它旨在评估出生后需要转运的 HIE 婴儿的 aEEG 是否可行，并且可以提供可读的临床信息。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Stratus EEG Centrum, Acquire Pro Software for 
			Microsoft Windows</td>
			<td>Stratus Software Solutions LLC 
			Kvikna Medical Lyngh&#225;ls 9 110 Reykjavik Iceland</td>
			<td>Version 4.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Trackit T4PCU24+8</td>
			<td>Lifelines Neuro 
			7 Clarendon Court 
			Over Wallop 
			StockBridge 
			Hants, UK</td>
			<td>SN: T4-170046 
			Issue: 4 C169</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra Subdermal Needle Electrode</td>
			<td>Natus 
			3150 Pleasant View Road 
			Middleton 
			WI 53562 
			USA</td>
			<td>019-476600</td>
			<td>14 mm x 0.38 mm 
			SST Needle 
			DIN 42802 connector</td>
		</tr>
	</tbody>
</table>

amplitude-integrated eeg in infants at risk of hypoxic-ischemic encephalopathy: a feasibility study in road and air transport in western australia

有 HIE 风险的婴儿需要早期识别并开始治疗性低温 （TH）。早期 TH 治疗与更好的结果相关。在决定是否开始 TH 时经常使用 aEEG。由于这通常仅限于三级中心，如果婴儿需要转运到提供 TH 的中心，TH 可能会延迟。我们旨在提供一种振幅集成脑电图 （aEEG） 的应用方法，并确定在运输过程中获取具有临床意义的信息的可行性。所有 ≥35 周，转诊时有 HIE 风险的婴儿均符合纳入标准。头皮电极置于 C3-C4 中;P3-P4 位置位于婴儿的头皮上，并连接到 aEEG 放大器。反过来，aEEG 放大器连接到带有 EEG 软件的临床平板电脑，以收集和分析 aEEG 信息。首席研究员和两名独立审查员 （盲法） 审查了背景痕迹和伪影的录音。预定义的数据质量标准被设置为移动伪影和软件阻抗通知。医护人员和家长完成了调查，以确保可接受性和易用性。

在 2022年 9 月 1 日至 2023年 6 月 5 日期间，25 名婴儿在西澳大利亚被运送，可能患有 HIE，并符合纳入资格。其中，20 名婴儿获得同意、招募并在运输过程中成功应用 aEEG。共有 5 名患者未招募，3 名患者因同意而错过。在一个案例中，该设备已启动自动软件更新并且无法使用;在一个案例中，婴儿没有 HIE 的风险，被认为没有资格招募。在这 20 名婴儿中，12 名 （12/20;60%） 为男性，8 ?...

作者聚焦：评估在新生儿转运过程中使用振幅集成脑电图的可行性

在这里，我们提出了一项可行性研究，以评估便携式振幅集成脑电图 （aEEG） 记录系统在转运疑似缺氧缺血性脑病 （HIE） 婴儿过程中的作用。

有缺氧缺血性脑病风险婴儿的振幅整合脑电图：西澳大利亚公路和航空运输的可行性研究

Infants at risk of HIE require early identification and initiation of therapeutic hypothermia (TH). Earlier treatment with TH is associated with better ...

amplitude-integrated-eeg-infants-at-risk-hypoxic-ischemic

Research

JoVE Journal

Medicine

Amplitude-Integrated EEG in Infants at Risk of Hypoxic-Ischemic Encephalopathy: A Feasibility Study in Road and Air Transport in Western Australia

377 次观看.  Government of Western Australia, Child and Adolescent Health Service. 我们想评估在新生儿转运过程中使用振幅积分脑电图的可行性。使用临床标准，我们确定了在运输过程中获得的痕迹是否可读。这有助于临床医生诊断和预后可能患有出生窒息或缺氧缺血性脑病后需要转运的婴儿。远程监控自动化设备和创新的运输系统，以改善生病和早产儿的预后。在运输过程中进行任何研究都有其困难，但对于改善护理至关...

视频: 作者聚焦：评估在新生儿转运过程中使用振幅集成脑电图的可行性

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 ...

科研

医学

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Cardiac Magnetic Resonance Imaging at 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher magnetic field strengths and potentially provides improved signal contrast and spatial resolution. While promising results have been achieved, ultra-high field CMR is challenging due to energy deposition constraints and physical phenomena such as transmission field non-uniformities and magnetic field inhomogeneities. In addition, the magneto-hydrodynamic effect renders the synchronization of the data acquisition with the cardiac motion difficult. The challenges are currently addressed by explorations into novel magnetic resonance technology. If all impediments can be overcome, ultra-high field CMR may generate new opportunities for functional CMR, myocardial tissue characterization, microstructure imaging or metabolic imaging. Recognizing this potential, we show that multi-channel radio frequency (RF) coil technology tailored for CMR at 7 Tesla together with higher order B0 shimming and a backup signal for cardiac triggering facilitates high fidelity functional CMR. With the proposed setup, cardiac chamber quantification can be accomplished in examination times similar to those achieved at lower field strengths. To share this experience and to support the dissemination of this expertise, this work describes our setup and protocol tailored for functional CMR at 7 Tesla.

The sensitivity gain inherent to ultrahigh field magnetic resonance holds promise for high spatial resolution imaging of the heart. Here, we describe a protocol customized for functional cardiovascular magnetic resonance (CMR) at 7 Tesla using an advanced multi-channel radio-frequency coil, magnetic field shimming and a triggering concept.

7号特斯拉的心脏磁共振成像

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher ...

Application of an Amplitude-integrated EEG Monitor (Cerebral Function Monitor) to Neonates

Amplitude-integrated EEG (aEEG) is an easily accessible technique to monitor the electrocortical activity in preterm and term infants in neonatal intensive care units (NICUs). This method was first used to monitor newborns after asphyxia, providing information about future neurological outcomes. The aEEG is also helpful to select newborns who benefit from cooling. The aEEG monitoring of preterm infants is becoming more widespread, as various studies have shown that neurodevelopmental outcome is related to early aEEG tracings. Here, we demonstrate the application of the aEEG monitoring system and present typical patterns that depend upon gestational age and pathophysiological conditions. Furthermore, we mention pitfalls in the interpretation of the aEEG, as this method requires accurate fixation and localization of the electrodes. Additionally, the raw EEG can be used to detect neonatal seizures or to identify aEEG application problems. In conclusion, aEEG is a safe and generally well-tolerated method for the bedside monitoring of neonatal cerebral function; it can even provide information about long-term outcome.

Application of an Amplitude-integrated EEG Monitor Cerebral Function Monitor to Neonates

Here, we show how to apply amplitude-integrated electroencephalography to monitor cerebral function in neonates.

振幅综合脑电图监测仪 (脑功能监护仪) 在新生儿中的应用

Amplitude-integrated EEG (aEEG) is an easily accessible technique to monitor the electrocortical activity in preterm and term infants in neonatal ...

An Anatomical Study of Nerves at Risk During Minimally Invasive Hallux Valgus Surgery

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional orientation and localization of structures that are not directly visible to the surgeon. This is especially critical for structures at risk like nerves or blood vessels. Optimization of the handling of cadaveric material and the combination of multiple techniques compensate for the limited availability of adequate specimens. The described protocol combines anatomical plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to help localize relevant structures, such as nerves, arteries, veins and to correctly position the portals during MIS procedures. Depiction of these structures in anatomy textbooks can differ from what is encountered in the surgical field; and for this reason, new anatomical studies with a surgical orientation are needed. However, this is a complex, time-consuming technique requiring specific training. The anatomical references described with the so-called 'clock method' provide the surgeon with an easy and reproducible system to locate the path of the nerves at risk in Hallux Valgus MIS procedures. This model can be extrapolated to many other minimally invasive surgical procedures.

Minimally invasive surgical (MIS) procedures rely on anatomical references to localize structures not directly visible to the surgeon. This manuscript describes a combined method of plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to locate the structures at risk during MIS procedures.

微创拇外翻手术中神经风险的解剖学研究

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional ...

Objective Nociceptive Assessment in Ventilated ICU Patients: A Feasibility Study Using Pupillometry and the Nociceptive Flexion Reflex

The concept of objective nociceptive assessment and optimal pain management have gained increasing attention. Despite the known negative short- and long-term consequences of unresolved pain or excessive analgosedation, adequate nociceptive monitoring remains challenging in non-communicative, critically ill adults. In the intensive care unit (ICU), routine nociceptive evaluation is carried out by the attending nurse using the Behavior Pain Scale (BPS) in mechanically ventilated patients. This assessment is limited by medication use (e.g., neuromuscular blocking agents) and the inherent subjective character of nociceptive evaluation by third parties.
Here, we describe the use of two nociceptive reflex testing devices as tools for objective pain evaluation: the pupillary dilation reflex (PDR) and nociception flexion reflex (NFR). These measurement tools are non-invasive and well tolerated, providing clinicians and researchers with objective information regarding two different nociceptive processing pathways: (1)&#160;the pain-related autonomic reactivity&#160;and (2) the ascending component of the somatosensory system. The use of PDR and NFR measurements are currently limited to specialized pain clinics and research institutions because of impressions that these are technically demanding or time-consuming procedures, or even because of a lack of knowledge regarding their existence.
By focusing on the two abovementioned nociceptive reflex assessments, this study evaluated their feasibility as a physiological pain measurement method in daily practice. Pursuing novel technologies for evaluating the analgesia level in unconscious patients may further improve individual pharmacological treatment and patient related outcome measures. Therefore, future research must include large well-designed clinical trials in a real-life environment.

Pain assessment in anesthetized patients who cannot communicate with the outside world in any way remains challenging despite the development of innovative objective pain evaluation tools. In this project, the pupillary dilation reflex and the nociception flexion reflex are assessed in critically ill, mechanically ventilated adult patients.

应用 Pupillometry 和伤害性屈曲反射的可行性研究--通气 ICU 患者的客观伤害评估

The concept of objective nociceptive assessment and optimal pain management have gained increasing attention. Despite the known negative short- and ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

大鼠胡萝卜动脉压力控制段气球损伤与腹腔ventit 治疗应用

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing (Neo)adjuvant Therapies

Surgery is often the first treatment for many solid tumors. However, local relapses frequently occur following primary tumor resection, despite adjuvant or neo-adjuvant therapies. This occurs when surgical margins are insufficiently tumor-free, resulting in residual cancer cells. From a biological and immunological perspective, surgery is not a null event; the wound healing environment is known to induce both pro- and anti-tumorigenic pathways. As a consequence, preclinical models for drug development aimed at preventing local relapse should incorporate surgical resection when testing new (neo)adjuvant therapies, to model the clinical settings in patients treated with surgery.
Here, we describe a mouse model of incomplete surgical resection of WEHI 164 soft tissue sarcoma that allows testing of (neo)adjuvant therapies in the setting of a wound healing response. In this model, 50% or 75% of the tumor is removed, leaving behind some cancer tissue in situ to model gross residual disease after surgery in the clinical setting. This model allows testing therapies in the context of surgery while also considering the wound healing response, which may affect the efficacy of (neo)adjuvant treatments. The incomplete surgical resection results in reproducible regrowth of the tumor in all mice in the absence of adjuvant therapy. Adjuvant treatment with checkpoint blockade results in reduced tumor regrowth. This model is thus appropriate for testing therapies in the context of debulking surgery and its associated wound healing response and can be extended to other types of solid cancer.

A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing Neoadjuvant Therapies

In this protocol, we describe a mouse model of incomplete surgical resection of soft tissue sarcoma for testing (neo)adjuvant therapies.

用于测试（新）辅助疗法的不完全切除软组织肉瘤的小鼠模型

Surgery is often the first treatment for many solid tumors. However, local relapses frequently occur following primary tumor resection, despite adjuvant ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

使用化学活检进行移植质量评估

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...

On-Site Sampling and Extraction of Brain Tumors for Metabolomics and Lipidomics Analysis

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still in need. In recent years, mass spectrometry has become a method of choice for untargeted profiling of discriminatory compound as potential biomarkers of a disease. Biofluids are generally considered as preferable matrices given their accessibility and easier sample processing while direct tissue profiling provides more selective information about a given cancer. Preparation of tissues for the analysis via traditional methods is much more complex and time-consuming, and, therefore, not suitable for fast on-site analysis. The current work presents a protocol combining sample preparation and extraction of small molecules on-site, immediately after tumor resection. The sampling device, which is of the size of an acupuncture needle, can be inserted directly into the tissue and then transported to the nearby laboratory for instrumental analysis. The results of metabolomics and lipidomics analyses demonstrate the capability of the approach for the establishment of phenotypes of tumors related to the histological origin of the tumor, malignancy, and genetic mutations, as well as for the selection of discriminating compounds or potential biomarkers. The non-destructive nature of the technique permits subsequent performance of routinely used tests e.g., histological tests, on the same samples used for SPME analysis, thus enabling attainment of more comprehensive information to support personalized diagnostics.

The manuscript presents on-site sampling of human brain tumors with solid phase microextraction followed by their biochemical profiling towards the discovery of biomarkers.

代谢组学和利皮多米分析脑肿瘤的现场采样与提取

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still ...