维也纳总医院和维也纳医科大学为维也纳肝血动力学实验室提供基础设施。维也纳肝血动力学实验室的前任成员和同事应该了解他们的宝贵意见，帮助不断改进我们机构的HVPG测量和血管肝活检方法。此外，我们感谢胃肠病学和肝病科的护士，他们是维也纳血动力学实验室的重要组成部分，并不断为患者提供出色的护理。

此处描述的议定书符合维也纳医科大学人类研究伦理委员会的准则。
1. 准备工作
<ol><li>使用配备数字 X 射线荧光镜系统 （图 1A）、监视器系统 （图1B）、压力记录器和传感器 （图1C）连接到打印机或录制软件以及超声波设备 （图1D）的 X 射线专用/批准程序。此外，确保有足够的空间在患者席旁边有一个无菌工作区（图1E）。</li>
	<li>通过测量HVPG来评估疑似晚期慢性肝病（ACLD）患者。特别是，有以下特征之一的患者可能会接受HVPG测量:血小板减少和150 G/L，横截面成像的端索辅助材料，胃食管变异在启动非选择性β阻断剂治疗之前，阿西特，和肝肿瘤，计划切除。</li>
	<li>在手术中使用以下禁忌症:（一）由于血管或骑兵静脉血栓形成而无法获得血管，（二）临床上明显的出血性疾病（如传播血管内凝血）、（iii）反向肺炎和（iv）显著心律失常。</li>
	<li>在被告知手术的目的和潜在风险后，确保患者被固定并给予书面知情同意。</li>
	<li>说明必须在没有全身麻醉的情况下执行该程序。然而，低剂量的米达佐兰（高达0.02毫克/公斤体重）可能会提供给焦虑的病人。</li>
	<li>使用标准监测系统，通过非侵入性动脉血压测量、脉搏氧化和心电图来监测患者的生命体征。</li>
	<li>在压力袋中使用无菌盐水溶液冲洗压力传感器集。</li>
	<li>校准压力传感器（如果不预先校准，在开始测量之前根据外部压力参考校准，例如使用 13.6 厘米 H2O 等于 10 mm Hg 的水柱）。</li>
	<li>将传感器放在右中庭的水平，瞄准患者的中轴线。</li>
	<li>准备压力记录器/记录软件，以便记录压力跟踪。</li>
	<li>确保所有必要的无菌设备都准备好了（见 材料表）。</li>
	<li>要求病人在病人的长凳上占据一个超高的位置。</li>
	<li>让操作员清洗和消毒她/他的手和前臂。</li>
	<li>戴上手术帽和面罩，无菌手套和无菌外套。</li>
	<li>使用无菌表盖，为中央静脉通道所需的设备准备无菌工作区（图 2A）。</li>
</ol>2. 无菌条件下的中央静脉进入
<ol><li>指示患者将头稍微转向左侧，然后用消毒剂溶液对右前颈和侧颈区域进行消毒。</li>
	<li>将覆盖皮肤的无菌毛巾放在胡萝卜三角形周围。</li>
	<li>使用超声波探针（由无菌的美国探头盖护套覆盖）来识别正确的内部血管和适当的穿刺部位。</li>
	<li>用21G针渗透随后血管进入区域，并应用局部麻醉剂（利多卡因2%，5 mL）。然后，等待1-2分钟的全部效果。</li>
	<li>准备中央静脉通道所需的所有材料（见 材料表）。</li>
	<li>使用 7.5 F 导管导管护套设备，使用超声波引导将针头放在正确的内部血管中，并使用 Seldinger 技术（图 3A）通过针推进导线。 注:如果在HVPG测量后进行血管活检，则必须使用直径较大的10F肝活检介绍套（额外使用18G针头和导线）。</li>
	<li>使用刀片 11 手术刀沿着导线进行 2-3 毫米切口，以方便引入装有扩张器的血管通道护套。</li>
	<li>将装有扩张器的血管通路垂直护套插入导线的内部血管静脉中。</li>
	<li>取出血管通道护套的导线和扩张器。</li>
	<li>确保血管进入护套保持稳定的血管内位置，并将血管进入护套的注射端口定向到侧向（图3B）。 注意:不需要安装或录音。</li>
</ol>3. 将气球导管放置在肝静脉中
<ol><li>准备将气球导管放入肝静脉所需的所有材料（图2B）。</li>
	<li>用对比介质（染料）冲洗气球导管，通过重复的窒息/通缩检查气球的完整性，导管尖浸入盐水溶液中。不应发生气泡。 注意:根据导管本身在 X 射线上的可见性，您也可以仅使用无菌盐水或稀释染料来冲洗导管的血管流明。</li>
	<li>用无菌盐水溶液润湿外面的气球导管，然后将其插入血管通道护套。</li>
	<li>在荧光引导下将气球导管推进到劣质卡瓦静脉 （IVC）中。将气球导管的尖端对准患者背部，导管的轻微旋转通常允许从右中庭推进到 IVC。在某些情况下，指示患者深吸气可能会有所帮助（图3B）。</li>
	<li>将气球导管从 IVC 推进到肝静脉中。尝试通过反复移动与气球导管的尖端面向右向肝静脉和 IVC（图 3C）交界处的可疑区域获得肝静脉。</li>
	<li>确保导管进入一个稳定的位置，允许在2-4厘米的距离内反复测量自由肝静脉压力（FHVP），并有足够的空间在肝静脉的流明膨胀气球，以记录楔形肝静脉压力（WHVP）。</li>
	<li>检查通过充气气球（约2 mL的空气进入气球流明）和对比剂注射（约5 mL进入血管流明）的静脉（楔形位置）的充分遮挡，直到肝静脉消失到膨胀的气球可视化（图3D）。</li>
	<li>观察对比介质的停滞状态，并排除因气球遮挡静脉流明不足或静脉通信存在而冲刷的对比介质。如果观察到显着的对比度介质冲洗，请尝试重新定位气球导管。</li>
	<li>放气气球，用盐水冲洗导管的流明。</li>
</ol>4. 用于评估HVPG的血动力学读数
<ol><li>使用输液线将气球导管的血管流明连接到压力传感器。</li>
	<li>开始记录FHVP与气球的尖端2-4厘米从肝静脉的开口到IVC。曲线的波形必须稳定，不会随着时间而变化。 注:稳定值通常在15岁以后获得。</li>
	<li>充气气球，并继续记录WHVP，直到测量成为一个稳定的水平线，随着时间的推移没有变化。 注:楔形压力的稳定跟踪通常在 >40 s 之后获得。</li>
	<li>重复 FHVG （+15 s） 和 WHVP （+40 s） 的测量至少三次，以获得三倍的高质量读数 （图 3E）。 注:如果单个 FHVP/WHVP 读数之间观察到≥2 mm Hg 存在显著差异，则应进行其他测量。注意潜在人工制品的原因，如咳嗽、移动或在血动力学记录的相应时间点说话。</li>
	<li>记录在 IVC 中的压力在肝静脉的骨质水平以及正确的体外压力 （RAP） 的水平。</li>
	<li>停止录制。</li>
	<li>取出气球导管。</li>
	<li>计算 HVPG（从 WHVP 中减去的 FHVP）为 3 个测量的平均值。 注:如果在按顺序获得的 HVPG 读数之间注意到≥2 mm Hg 的显著差异，则应进行其他测量。</li>
	<li>继续进行血管外活检（步骤 5.1），或从内部血管中取出导管导管护套。 注:插入僵硬的导线可能有助于达到相同的肝内位置，用于随后的血管内肝脏活检。</li>
	<li>使用无菌纱布在内血管静脉血管插入部位顶部施加压力至少5分钟。</li>
</ol>5. 做好血管外肝活检的准备
注:两种不同的活检方法可用于获得血管外肝脏活检:渴望（第6步）或核心活检（第7步）。首先决定使用哪个系统，然后选择适当的活检针介绍护套，然后再进行步骤 5.1 到 5.3）。或者，核心TJBLX集的针头也可以插入到愿望集（注意:使用适当的直径），这导致一个更灵活的核心活检系统，可能更容易引入肝静脉（注意:此方法在这里没有详细描述）。
<ol><li>准备相应的血管肝活检 （TJLBX） 集 （渴望TJLBX:图 4A-B:核心 TJLBX:图 4C:请参阅材料表）。</li>
	<li>用无菌盐水冲洗活检针引针护套，或在血管内呼吸肝活检的情况下，用对比剂进行更好的可视化。</li>
	<li>使用与 3.4 中描述相同的技术，将活检针引入护套推进到肝静脉中。</li>
	<li>根据所使用的活检集，继续为吸入TJBX踩6.1步，或为核心TJBX步骤7.1。</li>
</ol>6. 血管内吸肝活检
<ol><li>使用 10 mL Luer 锁注射器用无菌盐水冲洗吸入性 TJLBX 针头，但在注射器中留下约 3 毫升，以方便吸入。</li>
	<li>通过活检针引入器护套轻轻推进活检针，直到针尖到达介绍者护套的末端。 注意:在推进针头时避免任何力或快速运动。要求患者深呼吸会降低 IVC 和肝静脉之间的角度，以便更容易地推进活检针。</li>
	<li>请病人屏住呼吸。</li>
	<li>使用 10 mL Luer 锁注射器应用吸力，将针头推进肝脏帕伦奇马。</li>
	<li>在仍然使用吸力（负压）时缩回针头。</li>
	<li>建议患者继续正常呼吸。</li>
	<li>取出针头（但不是针头介绍护套），并收获肝脏样本。 注:肝脏样本通常保留在注射器中，而不是针头（图4D）中。</li>
	<li>重复步骤 6.1 到 6.6 额外的针通道，直到获得足够的肝脏标本。</li>
	<li>在导管引入器护套上注入 5-10 mL 的对比介质，以排除肝胶囊穿孔。</li>
	<li>取出活检针引物护套。</li>
	<li>取出 10 F 肝活检套管，使用无菌纱布在内血管血管的血管插入部位顶部施加压力至少 5 分钟。</li>
</ol>7. 血管核心肝活检
<ol><li>通过拉握来加载核心 TJLBX 针头，直到拍摄机制锁定。通过活检针引入器护套轻轻推进核心活检针，直到针尖接近介绍者护套的末端。 注意:在推进针头时避免任何力或快速运动。</li>
	<li>请病人屏住呼吸。</li>
	<li>将针头推进肝脏帕伦奇马。</li>
	<li>通过扣动射击机制的扳机进行核心活检。</li>
	<li>建议患者继续正常呼吸。</li>
	<li>取出针头（但不是针头介绍护套），并收获肝脏样本。小针可能有助于去除肝脏样本（图4D）。</li>
	<li>重复步骤 7.1 到 7.6 额外的针通道，直到获得足够的肝脏标本。</li>
	<li>将5-10 mL的对比介质注射到导管引入器护套的侧端口，以排除肝胶囊穿孔的可能性。</li>
	<li>取出活检针引物护套。</li>
	<li>取出10F肝活检套管，使用无菌纱布在血管内静脉血管插入部位顶部施加压力约5分钟。</li>
</ol>

托马斯·雷贝格尔得到了库克医疗的赠款和物质支持，以及W.L.戈尔和协会讲座的资助和荣誉。菲利普·施瓦布尔、马库斯·佩克-拉多萨夫列维奇和迈克尔·特劳纳对于这份手稿没有什么可透露的。马蒂亚斯 · 曼多弗因从 W. l. 戈尔和同事的讲座而获得荣誉。

虽然HVPG测量需要大量的资源和训练有素的个人与干预技能和专业知识的阅读压力跟踪，它改善预测，并可能指导治疗决定，从而促进个性化的医学。此外，在同一会话中，通过跨性别途径安全获得肝脏活检标本的机会是赞成在三级护理中心实施肝血动力学实验室的另一个论点。事实上，指引支持在拥有足够专业知识和资源的中心使用HVPG测量技术。手术的安全性主要与血管进入内血管有关。一旦正确放置，剩余手术的风险可以忽略不计，如果将气球导管放置在肝静脉中的时间比预期的要长，患者的舒适度大多受手术时间的限制。虽然最近发表的一项有关患者报告结果的研究表明，HVPG程序是耐受性良好的41，低剂量的中佐兰止血（高达0.02毫克/千克体重）可用于缓解焦虑和促进患者舒适42。然而，一般麻醉和/或深麻醉与异丙酚和雷米芬他尼不能用于HVPG测量，因为这会影响血动力学读数43.
如果右内血管不能用于静脉进入（例如血栓形成），则可以改用左内血管或替代血管。重要的是，HVPG也可以通过股骨静脉测量，因为使用专门的导管。
然而，HVPG 测量的最关键部分是在气球充气和放气时正确记录压力跟踪，确保适当的放置和足够的时间，以获得正确的 WHVP 和 FHVP。然而，在导管的"自由"位置获得的波形可能会受到心跳或三叉星瓣在系统内回流的轻微影响，但仍应遵循直线。相比之下，气球充气时获得的"楔形"压力曲线应遵循前 3-5 s 的快速增加，然后在 10-30 s 上缓慢增加。最后，应获得一条反映正确鼻窦压力的直线和稳定线。在三次测量过程中发生的任何不一致情况应提示操作员获取额外的压力读数，以便识别正确的 FHVP 和 WHVP。
HVPG测量的并发症率很低，风险几乎完全与通常在正确的内部血管11，13进行的静脉接触有关。在手术前应与患者讨论的 HVPG 程序的常见副作用包括插入部位疼痛或轻微胸腔或腹部不适，当通过静脉卡瓦系统将导管推进到肝静脉和充气气球时。与静脉接触相关的特定但罕见的并发症包括进入地点的血肿、需要胸管的肺气管和心律失常。
HVPG测量的四个主要指标是:（一） 建立肝内门户高血压的诊断， （二） 通过诊断CSPH（HVPG ≥10 mm Hg）来识别有肝分化风险的患者;（iii） 指导静脉出血的初级或二级预防药理治疗，（iv） 评估部分肝切除术后肝衰竭的风险。虽然CSPH诊断和风险分层也可能通过成像/实验室研究或内窥镜检查2，44进行，目前，没有足够的替代方法来监测NSBB治疗的反应。虽然有些患者可能很容易被排除在主要肝切除考虑肝损伤的迹象（即腹腔或黄斑）45，HVPG的测量代表了术后发病率和死亡率的重要预测器，否则保存良好的肝功能（即补偿患者）46。
HVPG 与直接测量的门户压力（或更确切地说，整个肝脏的压力梯度）密切相关，正如它已显示为肝病的酒精和病毒病因9的患者。然而，某些肝脏疾病（例如结节再生增生）也可能影响前体抵抗力，影响门户高血压的严重程度，但HVPG47没有充分反映。此外，HVPG 也无法检测门户静脉血栓形成或门户静脉机械压缩引起的（额外）肝前门户高血压的存在。因此，腹部成像特别侧重于脾脏血管轴，在中性静脉，以及脾脏大小和腹腔的存在，应在不清楚的情况下，每当门户高血压的预肝成分被怀疑48。
最后，在进行皮下肝活检的禁忌症患者中，例如，由于遗传性或后天性出血性疾病或严重腹膜炎，使用血管外肝活检已被证明是安全的。此外，核心活检针的发展提高了TJBX51，52的诊断产量。在不明病因的急性肝衰竭的情况下，或在疑似肝硬化的情况下，出于安全原因，在大多数情况下，转体活检应优先于皮下肝活检。在减退患者中，皮下肝活检与严重出血或与手术相关的并发症风险相当大，而在补偿患者中，相关风险较低。不过，我们更愿意在所有肝硬化患者（包括补偿患者）中执行 TJBX，因为可以通过 HVPG 测量同时获得重要的预测信息。

肝硬化患者有出现并发症的风险，这些并发症大多与门户高血压（PHT）有关，如胃或食管静脉曲张1、2、3的腹水或出血。肝退补偿的风险与PHT2的程度有关。肝静脉压梯度（HVPG）的测量是估计肝硬化患者门户静脉压力的黄金标准，即评估鼻窦门户高血压4的严重程度。≥6 mm Hg 至 9 mm Hg 的 HVPG 表示门户压力升高（"亚线门户高血压"），而 HVPG ≥10 mm Hg 定义 CSPH。该协议详细描述了设备和流程，并突出了潜在的陷阱，并为故障排除提供了建议。
临床上，HVPG的测量表明（一）建立鼻窦门户高血压的诊断， （二）通过诊断CSPH（HVPG ≥10 mm Hg）来识别有肝病康复风险的患者:（三） 指导静脉出血的初级或二级预防的药理治疗，（四） 评估局部肝切除术后肝衰竭的风险。HVPG 被用作肝纤维化/功能改善和/或恶化的既定代孕标记，因为 HVPG 的减少转化为临床上有意义的益处5，而较高的 HVPG 值则与增加的静脉出血风险6有关。根据对非选择性β阻滞剂（NSBB）或病因疗法下患者HVPG变化的观察，HVPG减少10%被认为是临床上相关的7，8。
迄今为止，没有替代的非侵入性参数反映与HVPG类似的精度门户压力的程度。即使HVPG实际上是一种"间接"的方式来测量门户压力，它强烈相关，从而准确地反映"直接"测量的门户压力在肝硬化患者9。重要的是，HVPG测量应该使用气球导管进行，以最大限度地提高评估的肝帕伦奇马10，11，12。虽然HVPG测量是侵入性的，资源密集型的，需要干预技能和专业知识来解释压力读数的可靠性和合理性，但这种方法是目前诊断和监测肝硬化患者的门户高血压的黄金标准13，14，15。
简单的实验室值，如血小板计数，可能有助于估计CSPH的可能性。然而，血小板计数，或包括血小板计数的非侵入性分数，具有有限的预测值16。在肝硬化患者中显示 splomegaly17 或端粒体抵押品18 的成像模式表明存在 CSPH，但无助于量化门户高血压的实际程度。新型非侵入性成像工具，如肝脏19 和/或脾脏20 的弹性成像，对于排除或排除CSPH的存在是有用的。尽管如此，没有一种可用的方法能够直接测量门户压力21的动态变化。
一些具有里程碑意义的研究强调了HVPG的预测价值， 表明HVPG≥10毫米汞柱（即CSPH）对变异8的形成（以及与门户高血压22相关的并发症的发展）具有预测性，而HVPG的（药理学诱发的）减少则调节了变异生长23和减补偿7的相应风险。HVPG 反应是 NSB 在预防（经常性）静脉出血方面有效性的唯一既定替代。如果HVPG降低到12毫米汞≤值，或在NSBB治疗期间降低≥10-20%，患者可免受静脉出血和存活率增加24，25。同样，HVPG反应也降低了经补偿的肝硬化患者的阿西特和相关并发症的发病率5，26。多项研究已提供证据，支持使用HVPG指导疗法27、28、29、30、31、32。因此，在有足够经验的中心，HVPG反应可以指导治疗决策，为门户高血压患者提供个性化的药物。
此外，HVPG的测量可能作为概念验证研究的替代终点，评估从长凳到床边的肝硬化和/或门户高血压新疗法的有效性，如索拉芬尼布33，34，辛伐他汀35，36，牛磺酸37，或埃里卡桑38。最终，HVPG的测量也可以提供有关HCC39和肝衰竭后肝切除40的风险的重要预测信息。
在二级和三级护理中心应随时提供测量HVPG的基础设施。由于HVPG测量技术需要专门的培训和设备，因此学术和移植中心建立肝动力学实验室，促进门户高血压的最先进的诊断和管理，似乎是合理的。大容量中心每年执行数百次 HVPG 测量。根据我们的经验，在 50-100 次监督的 HVPG 测量之后，通常会获得足够的专业知识来进行准确的 HVPG 测量。

<table border="0" cellpadding="0" cellspacing="0" style="width:1628px;" width="1627">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:315px;">10 mL Luer-Lock syringe</td>
			<td style="width:335px;">Braun</td>
			<td style="width:317px;">REF 4617100V, LOT 17G03C8</td>
			<td style="width:661px;">Luer-Lock Syringe for connection with the aspiration biopsy set</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 mL Syringe 2x</td>
			<td>Braun</td>
			<td style="width:317px;">REF 4606108V</td>
			<td>Snonpyrogenic, nontoxic 10 mL syringe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 F liver biopsy&#160; introducer sheath set</td>
			<td>Cook Medical</td>
			<td style="width:317px;">REF RCFW-10.0-38, REF G07600</td>
			<td>Percutaneous Sheath Introducer Set (TJBX), 10F Port, 13cm, Check-Flo Performer Introducer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">18 G needle for biopsy introducer sheat</td>
			<td>Arrow International</td>
			<td style="width:317px;">REF AN-04318</td>
			<td>Introducer Needle for TJBX Set, 18 G, 6.35 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">21 G needle</td>
			<td>Henke Sass Wolf</td>
			<td style="width:317px;">REF 0086, Fine-Ject 21Gx2&#34;</td>
			<td>Sterile injeciton needle, 21 Gx2&#34;, 0.8 x 50 mm, for local anesthesia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3-way channels</td>
			<td>Becton Dickinson</td>
			<td style="width:317px;">BD Connecta Luer-Lok, REF 39402</td>
			<td>Three way channel with Luer-Lok connection system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7.5 F catheter introducer sheath set</td>
			<td>Arrow International</td>
			<td style="width:317px;">REF SI_09875-E</td>
			<td>Percutaneous Sheath Introducer Set, 7.5 F Port</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Aspiration TJLBX set</td>
			<td>Cook Medical</td>
			<td style="width:317px;">REF RMT-16-51.0-TJL, REF G20521</td>
			<td>TJ Liver Access and Biopsy Needle Set (Aspiration Set), 9 F-45 cm, 16 G-50.5 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Balloon catheter</td>
			<td>Gerhard Pejcl Medizintechnik Austria</td>
			<td style="width:317px;">REF 500765B</td>
			<td>Ferlitsch HVPG Catheter, 7F-65cm, Balloon:2.5mL, Pressure 50-90 kPa, GW: 0.032&#34;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blade 11 scalpel</td>
			<td>Medi-Safe Surgicals</td>
			<td style="width:317px;">MS Safety Scalpel REF /Batch 18012424</td>
			<td>11 blade safety scalpels, retractable, single use, 10 scalpels per package</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blunt tip fill needle</td>
			<td>Becton Dickinson</td>
			<td style="width:317px;">REF 303129</td>
			<td>Sterile blunt tip fill needle, single use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Contrast media (dye)</td>
			<td>Dr. Franz K&#246;hler Chemie GmbH, Bensheim, GER</td>
			<td style="width:317px;">ZNR 1-24112</td>
			<td>Peritrast 300 mg Iod/mL, 50 mL, contrastmedia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Core TJLBX set</td>
			<td>Cook Medical</td>
			<td style="width:317px;">REF RMT-14XT-50.5-LABS-100, REF G08285</td>
			<td>TJ Needle Introducer and Bx-Needle 7 F-53.5 cm, 14 G-53.5 cm/20 mm, 18 G-60 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Digital x-ray fluoroscope system</td>
			<td>Siemens</td>
			<td style="width:317px;">Model No 07721710</td>
			<td>ARCADIS Varic, mobile x-ray fluoroscope system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Disinfectant solution</td>
			<td>Gebro Pharma</td>
			<td style="width:317px;">1-20413</td>
			<td>Isozid-H</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Face mask</td>
			<td>MSP Medizintechnik GmbH</td>
			<td style="width:317px;">REF HSO36984</td>
			<td>Surgical face mask from double fleece, with tie, 50 pieces</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Guide wire for biopsy introducer sheat</td>
			<td>Arrow International</td>
			<td style="width:317px;">REF AW-14732</td>
			<td>Marked Spring-Wire Guide, TJBX Set, 0.032&#34;, 0.81 mm, 68 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Infusion line</td>
			<td>Rosstec Medical Products b Cardea GmbH &#38; Co</td>
			<td style="width:317px;">REF 220010, 100m</td>
			<td>Infusion line, Luer-Lok for connection of balloon catheter and pressure transducer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lidocaine 2%</td>
			<td>Gebro Pharma</td>
			<td style="width:317px;">Xylanaest, ZNR 17.792, 20mg/1mL (2%)</td>
			<td>Sterile vials Xylanest including 2% Lidocain hydrochloride for local anesthesia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Midazolame</td>
			<td>Roche</td>
			<td style="width:317px;">Dormicum, ZNr 1-18809, Midazolam 5mg/5mL</td>
			<td>Sterile vials Dormicum including Midazolam for sedation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monitor system</td>
			<td>Datex-Ohmeda by GE</td>
			<td style="width:317px;">Type F-CMREC1</td>
			<td>Patient monitoring system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Patient bench</td>
			<td>Silerlen-MAQET</td>
			<td style="width:317px;">Model No 7474.00A</td>
			<td>Mobile patient bench for x-ray fluoroscopy</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pressure bag</td>
			<td>Ethox Corp</td>
			<td style="width:317px;">REF 4005</td>
			<td>Pressure Infuser Bag 500 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pressure recorder</td>
			<td>Edwards Lifesciences</td>
			<td style="width:317px;">Ref T001631A, Lot 61202039</td>
			<td>TruWave 3 cc/150 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pressure transducer</td>
			<td>Edwards Lifesciences TM</td>
			<td style="width:317px;">REF T001631A</td>
			<td>Pressure Monitoring Set (1x), 3 cc/150 cm, TruWave TM</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Recording software&#160;</td>
			<td>Datex Ohmeda by GE</td>
			<td style="width:317px;">Software S/5 is property of Instrumentarium Corp of Datex-Ohmeda TM</td>
			<td>Datex-Ohmeda S/5TM Collect - Software to record pressure tracings of the patient monitor system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile coat</td>
			<td>Lohmann &#38; Rauscher International GmbH &#38; Co</td>
			<td style="width:317px;">REF 19351</td>
			<td>Surgical Gown, Different sizes, e.g. L-130 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile gauze</td>
			<td>Hartmann</td>
			<td style="width:317px;">REF 401798, 10x10cm gauze</td>
			<td>10 x 10 cm sterile gauze, 10 pieces per package</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile gloves</td>
			<td>Meditrade</td>
			<td style="width:317px;">REF 9021</td>
			<td>Gentle Skin sterile gloves, different sizes, e.g. 7.5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile saline solution</td>
			<td>Fresenius Kabi</td>
			<td style="width:317px;">NaCl 0.9%, B009827 REV 03</td>
			<td>Physiologic Saline Solution 0.9% NaCl, 309 mosmol/L, pH-Wert: 4.5-7.0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile saline vessel</td>
			<td>KLS Martin</td>
			<td style="width:317px;">REF K8A,&#160; 18/10 Jonas</td>
			<td>Sterilizable Metal Vessel for sterile saline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile table cover</td>
			<td>Hartmann</td>
			<td style="width:317px;">REF 2502208</td>
			<td>Table Cover, Foliodrape 150 x 100 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile towel</td>
			<td>BARRIER by M&#246;lnlycke Health Care</td>
			<td style="width:317px;">REF 706900</td>
			<td>Adhesive OP-Towel, 100 x 100 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile US probe cover sheath</td>
			<td>Websinger</td>
			<td style="width:317px;">REF 07014</td>
			<td>Sterile ultrasound probe cover, 20 x 60 cm, inluding two sterie adhesive tapes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stiff guidewire</td>
			<td>Cook Medical</td>
			<td style="width:317px;">REF TSMG-35-180-4-LES, G46729</td>
			<td>Lunderquist Extra Stiff Wire Guide, 0.035&#34;-.89 mm, 180 cm, 4 cm flexible tip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical cap</td>
			<td>BARRIER by M&#246;lnlycke Health Care</td>
			<td style="width:317px;">REF 620500, PCS 100, Colour Green</td>
			<td>Surgical Cap</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ultrasound device</td>
			<td>FUJIFILM SonoSite</td>
			<td style="width:317px;">Model M-Turbo, REF P17000-17</td>
			<td>Mobile ultrasound system</td>
		</tr>
	</tbody>
</table>

measurement of the hepatic venous pressure gradient and transjugular liver biopsy

在这里，我们提供了一个详细的协议，描述肝静脉压梯度（HVPG）测量在晚期慢性肝病患者的临床过程，然后指导血管活检。在局部麻醉和超声波指导下，导管引入器护套放置在正确的内部血管中。使用荧光引导，气球导管被推进到劣质的静脉卡瓦（IVC），并插入一个大的肝静脉。通过注射对比介质来保证导管的正确和足够的楔形位置，同时气球会阻断罐式肝静脉的外流。校准外部压力传感器后，通过楔形肝静脉压力 （WHVP） 和自由肝静脉压力 （FHVP） 的三重记录获得连续压力记录。FHVP 和 WHVP 之间的差异称为 HVPG，值≥10 mm Hg 表示临床上显著的门户高血压 （CSPH）。在取出导管之前，记录 IVC 中在同一级别获得的压力读数以及正确的体外压力。
最后，可以通过相同的血管途径进行血管外肝活检。不同的系统可用:然而，核心活检针比吸入针更受欢迎，尤其是对于心律失常的肝脏。同样，在荧光镜指导下，活检针引体护套被推进到肝静脉中。接下来，通过介绍者护套轻轻推进血管活检针:（i） 在吸入活检的情况下，针头在吸入下进入肝帕伦奇马，然后快速取出，或（ii）在核心活检的情况下，切割机制在帕伦奇马内触发。可以通过变性活检安全地进行几个单独的通道，以获得足够的肝脏标本。在有经验的手中，这些程序的组合大约需要30-45分钟。

在肝功能保存完好的补偿患者（即没有任何肝退化史，如腹腔或静脉出血）中，测量的HVPG值可能是正常的，或在亚线门户高血压（HVPG 6-9 mm Hg）范围内。然而，补偿的患者可能会发展CSPH（HVPG≥10毫米汞柱），这表明开发变异或肝退补偿的风险增加。反过来，食管或胃静脉曲张的患者，HVPG通常在CSPH的范围，如果不是患者应评估门户静脉血栓的存在或其他原因的预肝/前体门户高血压。在有食管静脉曲张出血史的患者中，HVPG通常至少为≥12毫米汞柱。同样，在肝硬化引起的腹水患者中，即门户高血压腹水，HVPG值通常预计至少为≥10毫米汞柱（图3E）。≥20 mm Hg 的 HVPG 值表示未能控制出血和/或复发性变异出血的高风险，如果患者出现静脉出血，应成为先发制人的跨血管内端粒细胞分流 （TIPS） 置换的候选者。虽然高发性肝硬化和严重门户高血压患者的 HVPG 值可能上升到 +30 mm Hg，但 hVPG 值 =40 mm Hg 非常不寻常，应严格评估测量的准确性和压力跟踪。
FHVP 和 IVC 的极高值可能暗示压力传感器校准不正确，但也可能表示右心力衰竭或三聚氰胺瓣膜反转。FHVP 和 IVC 压力之间超过 ≥4mm Hg 的差异暗示了肝静脉或巴德-基亚里综合症的外流阻塞/狭窄。然而，在严重的巴德-基亚里综合征与肝静脉完全血栓阻塞的情况下，气球导管的插入通常是不可能的。因此，如果怀疑有巴德-基亚里综合症或其他肝外流阻塞的原因，建议对肝静脉进行多普勒超声波检查。
如果在气球充气时在对比介质喷射过程中观察到分流，则通常低估 HVPG。然而，虽然在这种情况下，HVPG的绝对值不能用于估计预后或指导药理治疗，但如果HVPG记录在10毫米汞柱≥，CSPH的诊断仍然可以进行。
在静脉出血的初级预防中，对非选择性β阻塞治疗的血动力学反应（例如 与基线相比，每天切开12.5毫克）被定义为HVPG至少下降≥10%，或作为绝对值的降低-lt;12 mm Hg:例如，患者A（谁没有前一发作的静脉出血）有20毫米汞柱的基线HVPG。在施用雕刻醇 12.5 毫克/天 4 周后，记录了第二个 16 mm Hg 的 HVPG（雕刻）。因此，HVPG 减少了 4 mm Hg（即基线 HVPG 的 20%），表明血动力学响应。
在静脉出血的二次预防中，血动力学反应要求HVPG减少≥20%（或绝对值=lt;12 mm Hg）:例如， 患者B的基线HVPG为26毫米汞柱，丙醇为40毫克b.i.d.（80毫克累计日剂量），HVPG降至24毫米汞柱，仅下降8%，表明血动力学无反应。在没有血动力学反应的情况下，如果仅使用丙丙醇 40 毫克 b.i.d 治疗，患者 B 有很高的再生风险。
结果表明，HVPG指导药理治疗加内窥镜带结比NSBB的"不受控制"联合治疗加内窥镜带结结在二次预防中更有效，甚至提高了存活率32。
在记录HVPG后，可以使用相同的血管访问护套在同一会话中安全地进行血管肝活检。用血管吸入活检针获得的肝脏活检标本可能是支离破碎的，特别是在肝硬化患者中，因此，与用核心活检针获得的活检相比，其代表性较小。然而，在预期肝纤维化不太明显的患者中，吸入针活检具有良好的样本质量，通常获得直径较大的标本。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58819/58819fig1.jpg"> 图1:肝血动力学实验室所需的设备。（A） 房间专用/批准使用X射线与数字X射线荧光镜系统的程序。 （B） 生命体征监测系统，即心电图、非侵入性血压和氧饱和。 （C） 加压无菌盐水冲洗的压力传感器。 （D） 用于引导血管访问的超声波设备。 （E） 无菌工作区域。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58819/58819fig1large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58819/58819fig2.jpg"> 图2:HVPG测量所需的无菌设备。 此图显示了用 （A） 中央静脉通道所需的材料准备的无菌工作区域，以及气球导管 （B） 放置在肝静脉中所需的材料。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58819/58819fig2large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58819/58819fig3.jpg"> 图3:导管导管护套的放置和气球导管的正确定位。（A） 导管介绍器护套正确放置在正确的内部壶静脉中。 （B） 气球导管进入 IVC。 （C） 气球导管推进到右肝静脉。 （D） 气球导管与膨胀的气球和肝静脉可视化对比媒体注射。 （E） 代表自由和楔形肝压读数的压力跟踪。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58819/58819fig3large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58819/58819fig4.jpg"> 图4。血管肝活检所需的设备。（A） 10 F 转体肝活检套管。（B）跨血管吸气活检系统，包括活检针介绍护套和连接到 10 mL Luer-lock 注射器的血管吸入活检针。（C）跨血管核心活检系统，包括带侧端口的针头介绍护套和核心活检针。（D）通过血管活检获得的肝脏标本。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58819/58819fig4large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>

Watch this Scientific Journal Video about Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy at JoVE.com

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

在这里，我们提出了一个测量肝静脉压梯度（HVPG）的协议，这是诊断临床上显著门户高血压的黄金标准。此外，我们描述了如何在同一会话内进行血管内肝脏活检。

肝静脉压梯度和转性肝活检测量

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

measurement-hepatic-venous-pressure-gradient-transjugular-liver

Nanyang Technological University

Research

JoVE Journal

Medicine

19.1K 次观看.  Medical University of Vienna. 在这里，我们提出了一个测量肝静脉压梯度（HVPG）的协议，这是诊断临床上显著门户高血压的黄金标准。此外，我们描述了如何在同一会话内进行血管内肝脏活检。

视频: Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

科研

医学

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

部分原位肝移植大鼠模型的肝动脉重建的外科手术

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

肝转移的临床前小鼠模型

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory&#160;rate&#160;and heart&#160;rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.
This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41&#177;2.94 cmH2O which was significantly higher (P&#60;0.05) than before resection (6.87&#177;2.39 cmH2O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.

This film demonstrates how to acquire systemic and hepatic hemodynamics in mice. The whole monitoring includes acquisition of vital parameters, systemic blood pressure, central venous pressure, common hepatic artery flow rate, and portal vein pressure as well as the portal flow rate in mice.

全身及肝脏血流动力学参数的监测小鼠

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential phenotypic changes are best assessed by measurement of the changes in lung elasticity. To best understand specific mechanisms underlying such pathologies in mice, it is essential to make functional measurements that can reflect the developing pathology. Although there are many ways to measure elasticity, the classical method is that of the total lung pressure-volume (PV) curve done over the whole range of lung volumes. This measurement has been made on adult lungs from nearly all mammalian species dating back almost 100 years, and such PV curves also played a major role in the discovery and understanding of the function of pulmonary surfactant in fetal lung development. Unfortunately, such total PV curves have not been widely reported in the mouse, despite the fact that they can provide useful information on the macroscopic effects of structural changes in the lung. Although partial PV curves measuring just the changes in lung volume are sometimes reported, without a measure of absolute volume, the nonlinear nature of the total PV curve makes these partial ones very difficult to interpret. In the present study, we describe a standardized way to measure the total PV curve. We have then tested the ability of these curves to detect changes in mouse lung structure in two common lung pathologies, emphysema and fibrosis. Results showed significant changes in several variables consistent with expected structural changes with these pathologies. This measurement of the lung PV curve in mice thus provides a straightforward means to monitor the progression of the pathophysiologic changes over time and the potential effect of therapeutic procedures.

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung&#8217;s structural changes with developing pathology.

在小鼠肺压力容积曲线的测量

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic agent melphalan. Briefly, the procedure starts with the setup of a percutaneous veno-venous bypass from the femoral vein to the external jugular vein. Via a laparotomy, catheters are then inserted into the proper hepatic artery and the caval vein. The portal vein and the caval vein, both supra- and infrahepatically, are then clamped. The arterial and venous catheters are connected to a heart lung machine and the liver is perfused with melphalan (1 mg/kg body weight) for 60 min. This way it is possible to locally perfuse the liver with a high dose of a chemotherapeutic agent, without leakage to the systemic circulation.
In previous studies including patients with isolated liver metastases of uveal melanoma, an overall response rate of 33-100% and a median survival between 9 and 13 months, have been reported. The aim of this protocol is to give a clear description of how to perform the procedure and to discuss IHP as a treatment option for liver metastases of uveal melanoma.

Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

隔离肝脏灌注作为治疗葡萄膜黑色素瘤肝转移

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

Generation of a Humanized Mouse Liver Using Human Hepatic Stem Cells

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in which the liver has been repopulated with functional human hepatocytes, could serve as a useful tool for investigating human hepatic cell biology, drug metabolism, and other preclinical applications. One of the key factors required for successful transplantation of human hepatocytes into mice is the elimination of the endogenous hepatocytes to prevent competition with the human cells and provide a suitable space and microenvironment for promoting human donor cell expansion and differentiation. To date, two major liver injury mouse models utilizing fumarylacetoacetate hydrolase (Fah) and uroplasminogen activator (uPA) mice have been established. However, Fah mice are used mainly with mature hepatocytes and the application of the uPA model is limited by decreased breeding. To overcome these limitations, Alb-toxin receptor mediated cell knockout (TRECK)/SCID mice were used for in vivo differentiation of immature human hepatocytes and humanized liver generation. Human hepatic stem cells (HpSCs) successfully repopulated the livers of Alb-TRECK/SCID mice that had developed lethal fulminant hepatic failure following diphtheria toxin (DT) treatment. This model of a humanized liver in Alb-TRECK/SCID mice will have functional applications in studies involving drug metabolism and drug-drug interactions and will promote other in vivo and in vitro studies.

Here, we present a novel humanized mouse liver model generated in Alb-toxin receptor mediated cell knockout (TRECK)/SCID mice following the transplantation of immature and expandable human hepatic stem cells.

利用人体肝干细胞人源化小鼠肝脏生成

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

直接节段性肝内交付使用大鼠肝门部夹具模型方法

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells

Natural killer (NK) cells are a subset of the cytotoxic lymphocyte population of the innate immune system and participate as a first line of defense by clearing pathogen-infected, malignant, and stressed cells. The ability of NK cells to eradicate cancer cells makes them an important tool in the fight against cancer. Several new immune-based therapies are under investigation for cancer treatment which rely either on enhancing NK cell activity or increasing the sensitivity of cancer cells to NK cell-mediated eradication. However, to effectively develop these therapeutic approaches, cost-effective in vitro assays to monitor NK cell-mediated cytotoxicity and migration are also needed. Here, we present two in vitro protocols that can reliably and reproducibly monitor the effect of NK-cell cytotoxicity on cancer cells (or other target cells). These protocols are non-radioactivity-based, simple to set up, and can be scaled up for high-throughput screening. We also present a flow cytometry-based protocol to quantitatively monitor NK cell migration, which can also be scaled up for high-throughput screening. Collectively, these three protocols can be used to monitor key aspects of NK cell activity that are necessary for the cells&#39; ability to eradicate dysfunctional target cells.

Evasion of natural killer (NK) cell-mediated eradication by cancer cells is important for cancer initiation and progression. Here, we present two non-radioactivity-based protocols to evaluate NK cell-mediated cytotoxicity toward hepatic tumor cells. Additionally, a third protocol is presented to analyze NK cell migration.

肝肿瘤细胞背景下自然杀伤细胞介导细胞毒性和迁移的测量

Natural killer (NK) cells are a subset of the cytotoxic lymphocyte population of the innate immune system and participate as a first line of defense by ...