Lennart Wiman (photographer) is acknowledged for excellent video recording.

患者签署知情同意书，并研究在哥德堡大学批准的区域伦理审查委员会。 1.麻醉<ol><li>设置所有监测（心电图，血压袖带）17。 </li><li>插入一个硬膜外导管在较低到中等水平的胸椎17。 </li><li>诱导麻醉异丙酚2毫克/公斤，150微克芬太尼，罗库溴铵和0.6毫克/千克，通过一个预先插入外周静脉导管。 </li><li> Preoxygenate和插管病人。维持麻醉与七氟醚，氧气和空气17。 </li><li>插入双腔中心静脉导管和动脉导管。监测CVP和动脉压17。 </li></ol> 2.隔离容器的肝脏和插管<ol><li>管理100 IE /公斤肝素，并给予抗生素预防（ 如 160毫克的甲氧苄啶和800毫克磺胺甲恶唑的）。 </li><li>使用经皮Seldinger技术，将一个17星期五导管在股静脉和15神父导管在颈外静脉18。通过透视检查导管的位置。 </li><li>由一个L形切口，从剑突下中线进行剖腹，然后延伸到一权利肋下切口19。 </li><li>确认在腹部有一个完整的检查和触诊无肝外转移。 <ol><li>如果发现任何可疑病灶，冰冻切片检查恶性肿瘤。 </li><li>终止该程序是否有肝外表现（periaortal淋巴结肿大，腹部转移）的证据。淋巴结肿大的肝门不被视为禁忌。 </li></ol></li><li>通过将镰状韧带，左和右三角韧带和冠状韧带19调动肝脏。 </li><li>解剖腔静脉从静脉肾静脉开始了与腹膜后支流静脉结扎隔膜。 </li><li>确定右肾上腺静脉和放置血管环周围。 </li><li>确定正确的生殖腺静脉。剖析它的5-10厘米。广场周围的入口荷包缝合到腔静脉。 <ol><li>放置一个结扎远端的性腺静脉。用剪刀，从腔静脉打开生殖腺静脉3-5厘米。 </li><li>通过正确的生殖腺静脉插入一个18神父线增强导管放置尖retrohepatically上述肾静脉。 </li><li>周围放置性腺静脉结扎以固定导管的位置。 </li><li>验证的回血通过导管。 Heparinize导管18。 </li></ol></li><li>解剖肝十二指肠韧带和可视化的肝总动脉的胃十二指肠动脉和适当的肝动脉。 <ol><li>围绕所述共同肝ARTE地方容器循环RY和gastrodoudenal动脉。 </li><li>解剖门静脉和放置血管环周围。 </li><li>分离含有胆总管韧带的其余部分，并且包括在容器中循环这一切。 </li></ol></li><li>解除胃十二指肠动脉与容器循环。中断动脉流入后，用剪刀做一个切口。 <ol><li>插入8-12神父导管，其尖端插入肝固有动脉。安全与胃十二指肠动脉周围结扎的位置。 </li><li>确认回血在导管，然后heparinize它18。 </li></ol></li><li>素灌注系统与一个单元包装的红细胞，100毫升白蛋白（50克/升），加入100ml Tribonat（碳酸氢钠），2500单位的肝素（5000 IE /毫升）和250ml的晶体液。 </li><li>连接动脉和腔静脉导管的灌注系统，包括一个心脏肺机配备了一个滚子泵和加热器吨ogether一次性类型的静脉水库，氧合器和管道系统。 </li><li>连接股骨和外部颈静脉导管的静脉 - 静脉旁路泵组成的离心泵，以及一个加热器单元和启动旁路。 </li><li>通过拧紧围绕腔静脉infrahepatically以上肾静脉一个Rumel止血带和夹紧腔静脉suprahepatically使用Klintmalm血管钳完成肝脏的隔离。 <ol><li>紧绕胆总管，门静脉和右肾上腺静脉Rumel止血带，最后夹紧使用血管钳肝动脉。 </li></ol></li></ol> 3.灌注<ol><li>启动灌注和手动监控在静脉储血水平。调节流速直至稳态水平达到时，500-1,200毫升/分钟之间，通常的流速。 </li><li>将热敏电阻探测到左右肝叶和一个直接进入动脉导管。 </li><li>热灌注针对40 摄氏度的目标肝脏温度保持整个灌注这个温度。 </li><li>将闪烁探测器在静脉转流泵头上。 </li><li>校准泄漏监测系统，通过注射小剂量的10活度99m锝白蛋白（1毫升）进入体循环，并注册基线活性。 <ol><li>注入一个10倍高的剂量，100活度99m锝白蛋白（1毫升）中，进入肝灌注单元。 </li><li>记录从所述灌注系统的任何泄漏到全身循环如由闪烁探测器测量的增加的活性。 </li><li>记录从体循环到灌注系统如静脉储量增任何泄漏。 </li></ol></li><li>当建立的流量的稳态，无泄漏的存在和肝脏温度已达到40℃时，以一个剂量注入马法兰1毫克/公斤体重到灌注系统分成两个剂量给予30分钟分开。 </li></ol> 4.结束和灌注导管取出<ol><li>灌注的60分钟后，倒空从系统的灌注液和灌溉的肝脏用1,000ml的晶体。 </li><li>添加一个单位包装红细胞来灌注机组，注入到肝脏，然后停止灌注。 </li><li>松开Klintmalm夹，并从腔静脉的Rumel止血带。 </li><li>从肝十二指肠韧带松开止血带Rumel。 </li><li>从胃十二指肠动脉取出结扎。取出导管和结扎血管。 </li><li>释放来自肝动脉夹。 </li><li>从生殖腺静脉取出导管结扎血管。 </li><li>停止静脉 - 静脉转流泵和断开管。 </li><li>关闭腹部有一个不间断筋膜缝合，然后在皮内缝合皮肤。 </li><li>删除C在股骨和颈外静脉atheters和申请的局部压力为5分钟。 </li></ol> 5.术后护理<ol><li>拔管患者并传输到术后护理单元24小时。保持监测中央静脉压和动脉血压。 </li><li>病人转移到外科为典型的3-7天手术后的护理。 </li><li>继续与低分子量肝素（LMWH），在总共4周剂量的5000的IE /天。 </li></ol>

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The authors have nothing to disclose.

转移性葡萄膜黑色素瘤有几个治疗方案。化疗6，免疫治疗23和靶向治疗24一直没能表现出改进的操作系统。患者很少肝转移，肝切除术是一种选择12。其他治疗方法包括肝动脉灌注化疗（HIA）9，肝动脉化疗栓塞（TACE）25和选择性体内放射治疗（SIRT）26。 国际水文计划是一个重大而复杂的外科手术，并因为由Ausman的第一?...

葡萄膜黑色素瘤是成人中最常见的原发性眼内恶性肿瘤，与发病率是最高的高加索人群1。局部治疗包括眼球摘除或局部斑块放射治疗2,3。无论局部治疗，转移性疾病最终会发展约35-50％的患者4,5。肝脏是转移的最常见的部位与本孤立肝转移患者约50％。中位总生存期（OS）的肝转移患者为6-12个月4。几个不同的系统性和区域性的治疗策略进行了探讨，但生存率没有改善2。 治疗全身化疗，单质（达卡巴嗪或替莫唑胺6）或联合治疗（BOLD方案7，吉西他滨与苏消安8），显示了较少的应答率则的10％。肝动脉内化疗（HIA）已经呈现出稍高总反应率相比全身化疗，但不增加整体存活率9。化学栓塞（TACE）与顺铂和卡铂显示出与9个月10中位生存在57％的患者有部分反应。选择性体内放射治疗（SIRT）已经显示与7个月11的中位存活的患者的62％的部分的响应。在肝切除术的研究，包括255例患者，14个月的总生存报告;在患者那里一个微观完整切除是可能的76，生存27个月12。 隔离肝脏灌注（IHP）是一个区域性的治疗选择，最初是由Ryan和Ausman发展。该技术背后的基本原理是通过外科手术隔离和灌注肝脏用高剂量的化疗剂的同时避免系统性至xicity 13。此外，并发热疗介导的化疗剂的变化在肿瘤血流和细胞膜通透性14增加的摄取。完整的血管隔离证实使用锝99标记的人血清白蛋白15放射性示踪技术。回顾II期临床研究已经显示为14个月（26与12个月）的潜在生存获益为接受国际水文计划相比，时间最长的幸存者，在瑞典16葡萄膜黑色素瘤肝转移患者。 在这里，我们提出国际水文计划的技术与目前的结果和未来前景简短评论。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Wire re-inforced catethers</td>
			<td>Medtronic Inc, Minnesota, USA</td>
			<td></td>
			<td>12-18Fr</td>
		</tr>
		<tr>
			<td>St&#246;ckert S5</td>
			<td>Sorin Group, Mirandola, Italy</td>
			<td></td>
			<td>Heart-lung machine</td>
		</tr>
		<tr>
			<td>Rotaflow</td>
			<td>Maquet, Jostra Medizintechnik AG, Hirrlingen, Germany</td>
			<td></td>
			<td>Centrifugal pump</td>
		</tr>
		<tr>
			<td>HU 35</td>
			<td>Maquet cardiopulmonary AG, Hirrlingen, Germany</td>
			<td></td>
			<td>Heater unit</td>
		</tr>
		<tr>
			<td>Flexbumin 200 g/l</td>
			<td>Baxter Medical AB, Kista, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tribonat</td>
			<td>Fresenius Kabi AB, Uppsala, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin LEO, 5000 IE/ml</td>
			<td>LEO Pharma AB, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer Acetate</td>
			<td>Baxter Medical AB, Kista, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vasculosis</td>
			<td>Cis-Bio International, Gif-Sur-Yvette, France</td>
			<td></td>
			<td>99mTc-albumin</td>
		</tr>
		<tr>
			<td>MedicView</td>
			<td>SystemData AB, Gothenburg, Sweden</td>
			<td></td>
			<td>Leakage monitoring system</td>
		</tr>
		<tr>
			<td>Alkeran</td>
			<td>Aspen Europe GmbH, Bad Oldesloe, Germany</td>
			<td></td>
			<td>melphalan</td>
		</tr>
	</tbody>
</table>

isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma

分离出的肝灌注（IHP）是一个过程，其中肝脏手术分离并灌注有高浓度的化疗剂美法仑。简单地说，该过程开始于从股静脉向外部颈静脉经皮静脉 - 静脉旁路的设置。经由剖腹，导管被插入到适当的肝动脉和腔静脉。门静脉和腔静脉，既supra-和infrahepatically，然后夹紧。的动脉和静脉导管被连接到心脏肺机和肝脏灌注美法仑（1毫克/千克体重）为60分钟。这种方式是可能的局部灌注用高剂量的化疗剂的肝癌，无泄漏到全身循环。 在以前的研究，包括患者的葡萄膜黑色素瘤孤立的肝转移，中33-100％的总反应率和9日和13月之间，中位生存期，已经报道了。此协议的目的是为了给如何执行过程描述清楚，并讨论国际水文计划作为葡萄膜黑色素瘤肝转移治疗方案。

初次报告包含的马法兰带或不带TNF-α治疗的22例患者。结果表明，总有效率为62％，其中两例CR（10％），以及11个月20中位总生存期。随后的研究报道了单独使用马法兰29例患者治疗的结果。该结果是用62％的总响应包括完整的10％响应和12个月的21位生存很相似。 在一项研究Rizell。等 ，被列入20例葡萄膜黑色素瘤。将患者分为三个不同时代的时间，结果显...

Watch this Scientific Journal Video about Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma at JoVE.com

Isolated Hepatic Perfusion as a Treatment 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 ...

isolated-hepatic-perfusion-as-treatment-for-liver-metastases-uveal

Research

JoVE Journal

Medicine

13.3K 次观看.  Institute of Clinical Sciences. 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.

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

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 ...

科研

医学

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, ...

MRI-guided dmPFC-rTMS as a Treatment for Treatment-resistant Major Depressive Disorder

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial prefrontal cortex (dmPFC) in patients with major depressive disorder (MDD). Technicians used a neuronavigation system to process patient MRIs to generate a 3-dimensional head model. The head model was subsequently used to identify patient-specific stimulatory targets. The dmPFC was stimulated daily for 20 sessions. Stimulation intensity was titrated to address scalp pain associated with rTMS. Weekly assessments were conducted on the patients using the Hamilton Rating Scale for Depression (HamD17) and Beck Depression Index II (BDI-II). Treatment-resistant MDD patients achieved significant improvements on both HAMD and BDI-II. Of note, angled, double-cone coil rTMS at 120% resting motor threshold allows for optimal stimulation of deeper midline prefrontal regions, which results in a possible therapeutic application for MDD. One major limitation of the rTMS field is the heterogeneity of treatment parameters across studies, including duty cycle, number of pulses per session and intensity. Further work should be done to clarify the effect of stimulation parameters on outcome. Future dmPFC-rTMS work should include sham-controlled studies to confirm its clinical efficacy in MDD.

Here we outline the procedure for MRI-guided repetitive transcranial magnetic stimulation to the dorsomedial prefrontal cortex as an experimental treatment for major depressive disorder.

MRI引导dmPFC-rTMS治疗作为治疗难治性抑郁症

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial ...

Percutaneous Hepatic Perfusion (PHP) with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects. Previously, this was performed during open surgery with satisfying results, but morbidity and mortality related to the open procedure, prohibited a widespread application. Therefore, percutaneous hepatic perfusion (PHP) with simultaneous chemofiltration was developed. Besides decreasing morbidity and mortality, this procedure can be repeated, hopefully leading to a higher response rate and improved survival (by local control of disease). During PHP, catheters are placed in the proper hepatic artery, to infuse the chemotherapeutic agent, and in the inferior caval vein to aspirate the chemosaturated blood returning through the hepatic veins. The caval vein catheter is a double balloon catheter that prohibits leakage into the systemic circulation. The blood returning from the hepatic veins is aspirated through the catheter fenestrations and then perfused through an extra-corporeal filtration system. After filtration, the blood is returned to the patient by a third catheter in the right internal jugular vein. During PHP a high dose of melphalan is infused into the liver, which is toxic and would lead to life threatening complications when administered systemically. Because of the significant hemodynamic instability resulting from the combination of caval vein occlusion and chemofiltration, hemodynamic monitoring and hemodynamic support is of paramount importance during this complex procedure.

Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

In this manuscript, we describe percutaneous isolated hepatic perfusion with simultaneous chemofiltration as treatment for unresectable liver metastases. This procedure is performed under general anaesthesia in the angiosuite by an experienced team, consisting of an interventional radiologist, a clinical perfusionist and anaesthesiologist.

经皮肝脏灌注（PHP）与美法仑作为囿于肝脏不能手术切除转移的治疗

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn ...

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 ...

Treatment of Liver Metastases Using an Internal Target Volume Method for Stereotactic Body Radiotherapy

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For inoperable patients, local ablation therapies, such as stereotactic body radiotherapy (SBRT), can provide effective local tumor control with minimal toxicity. Because of its high precision and accuracy, SBRT delivers a higher radiation dose per fraction, is more effective, and targets smaller irradiation volumes than does conventional radiotherapy. In addition, steep dose gradients from target lesions to surrounding normal tissues are achieved using SBRT; thus, SBRT provides more effective tumor control and exhibits fewer side effects than conventional radiotherapy. The use of SBRT is prevalent for treating intracranial lesions (known as stereotactic radiosurgery); however, it is now also used for treating spinal and adrenal metastases. Because of advancements in image-guided assistance and respiratory motion management, several studies have investigated the use of SBRT for treating lung or liver tumors, which move as a patient breathes. The results of these studies have suggested that SBRT favorably controls tumors in the case of moving lesions.
Four-dimensional computed tomography (4D-CT) with an abdominal compressor (AC) is clinically convenient for effective respiratory motion management. Because this method is noninvasive and allows free breathing, its use reduces complications. Furthermore, patients consider this method convenient. Moreover, it is considered more efficient than other methods of respiratory motion management by physicians and therapists. The use of 4D-CT with an AC for treating pulmonary lesions has also been widely investigated, and the technique is gaining acceptance for treating hepatic lesions. However, the protocols for using 4D-CT with an AC for treating hepatic lesions are different from those used for treating pulmonary lesions. In this article, we describe a new protocol for SBRT with 4D-CT and an AC for treating liver metastases.

Stereotactic body radiotherapy (SBRT) requires rigorous accuracy and precision for delivering high radiation doses per fraction to small treatment volumes to improve tumor control and simultaneously reduce toxicity. Herein, we present a noninvasive and clinically convenient respiratory motion management protocol for SBRT for liver metastases.

内靶容积法治疗肝转移瘤的立体定向体放疗

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

体外常温肝灌注的小动物模型

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

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 ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

毛利精密切肝片作为肝脏生物学的Ex Vivo模型

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...