The authors acknowledge the financial support received from the state budget by the Slovenian Research Agency (program no. P3-0003, project no. J3-6796). The research was conducted in the scope of the EBAM European Associated Laboratory (LEA). This manuscript is the result of the networking efforts of the COST Action TD1104. We wish to thank Alenka Seliskar, Mojca Juvan (Veterinary Faculty, Clinic for Surgery and Small Animals) and Jakob Konda (Institute of Oncology, Ljubljana, Ljubljana, Slovenia) for all of their valuable help.

该协议被批准为农业，林业和食品部的伦理委员会，并遵循其准则。该协议生效80多个狗和猫20。 注:ECT治疗包括顺铂或博来霉素和肿瘤结节电脉冲曝光的管理。 1.病人选择注:患者选择允许医生确定患者是否是合适的或不从该治疗中获益。 <ol><li>评估患者的病史（约复发年龄，品种，性别和任何并发疾病的存在，肿瘤类型，随时间的肿瘤的演化，先前的治疗，其结果，复发和速度一般数据，拥有者的左右的成功性的期望的治疗）。 </li><li>执行所述患者的身体检查<ol><li>详细评估肿瘤为了给M如果潜在的侵入性肿瘤呈现治疗广泛的利润率（做出有关的大小决定:AKE一个可能的临床鉴别诊断和，以确定治疗区域执行用于细胞学评估和可能的组织活检细针抽吸用于组织学评估根据用于任何其他处理相同类型的肿瘤， 例如 ，肥大细胞瘤2R≤1cm处，1厘米余量应治疗的建议处理的边缘; R -半径）。 </li><li>评估以制备处理方法的结节的数量和大小。 </li></ol></li><li>一般检查<ol><li>为了使本病的分期和检查实现一个完整的一般检查: <ol><li>如果有可能排除了治疗，如果患者有任何慢性感染史（根据在一定区域流行病学）或无其它并发疾病免疫介导的疾病，可能会改变血液或coagulat离子变更（执行特定测试），如果动物是能够承受一系列深sedations和/或全身麻醉的，如果患者有顺铂的敏感性（标准剂量是60-70毫克/米2体表面积/ 21 d）或博莱霉素或已经接收博莱霉素累积剂量超过400.000 IU / 平方米 。 </li></ol></li><li>使用诊断成像技术（腹部的胸部和超声检查的X线），以便根据肿瘤类型来检查任何可能的转移。 </li></ol></li></ol> 2.实验室分析<ol><li>血汗工作<ol><li>执行完全血液学与（BUN），肌酸酐，丙氨酸转氨酶（ALT），碱性磷酸酶（AF）的血液尿素氮的生物化学面板和钙排除以确定晚期慢性肾脏和肝脏疾病和可能的最常见的副肿瘤综合征分析可具有一个EF临床阶段FECT的预后。 </li><li>做一个额外的具体血液分析: <ol><li>如果患者给予这有可能改变凝血能力的变化是什么药，检查凝血功能。 </li><li>如果使用侵入性治疗（针电极），检查血小板数;血小板的数目应&gt; 100或者在情况下，它是&lt;100，凝固轮廓（PT，APTT，纤维蛋白原，D-二聚体）应该是正常的。 </li><li>在根据肿瘤疾病和患者的临床状况的具体情况下，检查总蛋白，白蛋白，磷，钠（Na），钾（K），氯（Cl）的脂肪酶浓度。 </li></ol></li></ol></li><li>肿瘤（多个）组织学分析<ol><li>如果细胞学诊断先前进行组织学分析是没有必要的。在某些肿瘤类型中的情况下组织学分级是一个至关重要的预后因素（ 例如，肥大细胞瘤）进行苏木精和曙红站进不去16。 注:活检（切口或"冲"），可以ECT前的同一次访问中进行。在任何情况下，它是良好的临床实践来诊断肿瘤的类型。 注:此案充分评估进行深入讨论和澄清需要动物的主人之后。讲解给业主ECT什么是什么，可以从ECT在临床效果方面可以预期的。与业主讨论疗程:全身麻醉或深度镇静，治疗的总的持续时间，治疗的成本，复发的可能性下ECT会话的数量。 ECT的临床试验中获得患者纳入的情况下，业主的书面同意，如果病人是真正合格。 </li></ol></li></ol> 3.治疗方式的选择注:ECT犬使用顺铂和博莱霉素。对于ECT猫只使用博来霉素，顺铂是猫禁忌。 <ol><li&gt;治疗分开每种肿瘤。确定的位置和每个结节的大小及结节勾勒治疗方案的数量。 </li><li>使用公式V =ab²π/ 6（"a"是肿瘤结节的较大直径和"b"的肿瘤结节的垂直于"a"的直径）或V =abcπ/ 6（"计算肿瘤结节的体积一"，"b"和"c"的是肿瘤结节的三个垂直的直径）。 </li></ol> 4.工作形态的选择<ol><li>电极的选择 注:重要的是要选择最佳的电极类型电极配置会影响组织中的电场分布。两种类型的电极被使用:板和针状电极。板电极具有平行的不锈钢板:在电极之间的内部距离取决于肿瘤的大小和电脉冲的发生器的容量。一般的距离是吐温电极为6-8毫米。针状电极可被布置成行（平行阵列），或在六角形几何。六角形电极具有中心针和在圆形阵列6针。脉冲被圆形阵列中，并将它们与中心之一之间的电极之间。这些脉冲在每对电极之间的两个方向上施加。根据肿瘤结节的深度选择电极的种类和所需的处理区域中的整个提供适当的电穿孔。根据特定目标组织选择的电极。在一般情况下，这两种类型的电极可用于治疗。 </li><li>选择和准备顺铂或博来霉素 注:ECT由顺铂或博莱霉素或肿瘤结节电脉冲的照射静脉给药肿瘤内给药的。 <ol><li>选择合适的细胞抑制性和它的给药途径。 </li><li>溶解词splatin（顺式 - 顺铂II）中，如果以1mg / ml的浓度得到的粉末，在蒸馏水（否则药学可用于制备的溶液）。给顺铂瘤内。 </li><li>以3mg / ml的浓度溶解于生理盐水博莱霉素和给予肿瘤内或静脉内。 </li></ol></li><li>麻醉 注意:根据操作模态（ 表1）深度镇静或全身麻醉可以使用。 <ol><li>根据美国麻醉医师协会"（ASA）根据，因为可能的副肿瘤综合征的肿瘤类型的物理状态分类和评估病人。 </li></ol></li><li>深度镇静<ol><li>使用美托咪定（0.08毫克/千克），或在狗的双人组合美托咪定（0.025毫克/千克）与酒石酸布托啡诺（0.1毫克/千克），或按照特定的医院进行深度镇静的标准程序。 </li></ol></li><li>全身麻醉<ol> &lt;LI&gt; Premedicate与乙酰丙嗪（0.02毫克/千克）和美沙酮（2毫克/千克）。 </li><li>三十分钟后诱发全身麻醉硫喷妥钠（5毫克/千克）或丙泊酚（1毫克/千克）。 </li><li>放置气管内管和保持与经由T型件麻醉电路输送氧异氟烷麻醉。 </li><li>在麻醉以10ml / kg /小时和止痛的速率使用流体载体治疗哈特曼溶液与非类固醇止痛药（ 例如，卡洛芬2-4毫克/千克）的单剂量。 </li><li>使用眼药膏，以防止干燥时的麻醉下。治疗后，督促病人，直到它重新获得足够的意识胸骨斜卧。保持在诊所的病人约2至4小时。 注:在特定医院全身麻醉标准程序也可以遵循。 </li></ol></li><li>操作环境 注:工作人员进行培训，ECT运营商（DVM），麻醉师和助手。 <ol><li&gt;使用时，ECT下深度镇静进行任何救护室或手术室，其中全身麻醉是允许的。 </li><li>镇静和/或全身麻醉使用根据麻醉的程序所需的所有设备。 </li><li>对于ECT程序请参阅材料清单所有需要的材料。 </li></ol></li><li>操作员和周围的人才保障 注意:皮肤抗肿瘤药物的液滴很少曝光引起急性问题。顺铂和博来霉素不是通过完整的皮肤吸收。顺铂可能导致只能刺激。注入公司的结节或肿瘤时，可能会出现顺铂和博莱霉素泄漏。在针刺伤害的情况下，洗净用丰水和根据标准医学实践取伤口护理。 <ol><li>保护操作者和周围的助理口罩和手套。在皮肤暴露于顺铂或博来霉素溶液的情况下，立即大量用清水冲洗干净。 </li&gt; <li>根据有关抗癌药物的使用地方性法规，遵守如有质量保险计划设立的生物安全措施。 </li></ol></li><li>电脉冲发生器 注:电脉冲的应用可以与电脉冲的发生器来进行。通常，该装置是用集成软件和触摸屏界面来选择预先设计的电脉冲的方波脉冲发生器。它配备有由帘线连接到发生器的脉冲激活按钮的手持电极座。电脉冲的递送可以通过脚踏开关进行控制。电脉冲发生器应符合欧盟标准:2006/95 / CE从12/12/2006关于特定电压范围内使用的电子材料，2004/108 / CE从2004年12月15日关于电磁兼容性，并应符合FDA的规定。 </li><li>电脉冲的应用程序 注:钍电脉冲电子应用如下顺铂或博来霉素，博来霉素或静脉给药的肿瘤内给药。当给予肿瘤内细胞抑制剂应缓慢施加以防止泄漏。对于静脉内施用细胞抑制剂应当以大丸剂持续大约30秒被应用。的水基凝胶是用来实现在皮肤和电极，无论是板或针之间的良好接触，并实现较少异构电场分布。凝胶的导电性应为0.5至1个S /米17,18之间。所使用的凝胶的导电率为0.5的S /微米。平板电极应放置在该肿瘤在它们之间包围的方式。另外，为了达到电场应该在垂直方向上进行两次电脉冲的应用的更好分布。如果肿瘤小于电极之间的距离大，电极的应用程序应星吨从肿瘤的边缘（包括安全余量），并进展朝向肿瘤的中心。在针电极的情况下，应注意，所有的针插入组织以实现电场的适当分布。不需要凝胶的应用。否则，该过程是一样的在平板电极的情况下。 <ol><li>脱毛的皮肤中的电脉冲的应用领域</li><li>辖顺铂或博莱霉素。如果细胞生长抑制剂施用静脉内等待8分钟，并为电脉冲传递的时间间隔为20分钟。如果抑制细胞生长的瘤内给药开始的电脉冲的立即应用（注射后1分钟内）。 </li><li>电脉冲的应用程序之前，应用水基凝胶上脱毛皮肤。 </li><li>对于电极板使用这些脉冲参数:脉冲持续时间100微秒，幅度电极距离比1300 V /厘米，频率1赫兹或5千赫。递送8单极电脉冲的每次运行中的四个脉冲两列车，以1秒的时间间隔，在两个垂直方向。 </li><li>用于与行针电极使用的脉冲参数:脉冲持续时间100微秒，幅度到电极距离比1000伏/厘米，频率1或5千赫。单极电脉冲的每个运行由八个脉冲。 </li><li>对于针状电极，安排六角形的配置和使用脉冲参数:脉冲持续时间100微秒;六角形电极具有730伏的固定电压（如果电极之间的距离为7毫米; 1,040伏/厘米），以5kHz的频率和脉冲数取决于针的数量。在7针的情况下被应用24脉冲</li></ol></li><li>污染的材料的处理。 <ol><li>所有被污染的材料（注射器，针头，药瓶，纱布，手套）的用于焚烧容器进行处理。 </li></ol></li></ol>注:在每次访视，肿瘤结节用游标卡尺和拍照测量。这些访问还用于为了评价可能的局部和全身副作用。响应于该处理是基于"实体瘤的响应评价标准"（RECIST）19作为一个完整的响应（CR）的标准4周后得分-至少30％ -的结节，部分应答（PR）的消失降低目标病变的直径的总和，以作为参考基线总和的直径，进行性疾病（PD） - 至少在目标病灶的直径的总和增加20％，以作为参考的最小总和被研究，疾病稳定（SD） - 既不是足够的收缩以符合PR，也不足够增加晋级PD，作为参考的最小总和的直径。对于所有响应的定义至少需要4周的时间换曲alifying每种类型的反应。观测时间被计算为第一处理的日期和病人的最后检查的日期之间的时间间隔。 20在每次随访检查-可能的副作用是根据兽医肿瘤协作组毒性规模（CTCAE VCOG）测定。 <ol><li>预约1周治疗后。附表治疗2周后预约。安排治疗4周后预约。 4周后安排每月一次的约会。 </li></ol>

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The authors have nothing to disclose.

电化学已被证明是对不同的原发肿瘤或转移的狗和猫的高度有效的。电化学能治愈目的为孤或多个皮肤或皮下肿瘤结节或作为辅助治疗，以外科手术中使用。选择的几个肿瘤在狗，包括肥大细胞瘤，标准治疗是肿瘤的手术切除。手术比较在狗治疗肥大细胞瘤的标准治疗，以ECT的一项研究表明，ECT治疗导致的完全反应70％，而手术导致了完全反应50％11证明，认为ECT是同等待遇手术。 对于ECT的明显的抗肿瘤功效两个条件必须满足，这是协议中的关键步骤。第一个是细胞抑制剂的管理。在靶组织中的药物的存在下确定的ECT效率作为药物需要存在在细胞内以杀死它们。当瘤内注射药物，良好的组织保留的标志是组织美白。此外，可以通过在肿瘤中的针或期间进一步ECT会话坏死组织中产生的空穴发生在给药后的溶液一些泄漏。从而，达到1毫克/平方厘米3（顺铂）或3000的UI /厘米3（博来霉素）的浓度然后可以在这些情况下纯理论，但是，我们的目标是尽量保留尽可能多的溶液尽可能在靶组织中。此外，电脉冲应当尽可能快，以获得这有助于溶液保持一个血管收缩作用施加。 管理的两个不同的路线可以用ECT使用:瘤内和静脉。 ECT与瘤内顺铂或博来霉素或静脉注射博莱霉素的抗肿瘤效果相当于21，而ELEctrochemotherapy与顺铂的静脉应用是22，因此在顺铂瘤内注射仅用于事倍功半。顺铂药物的另一个重要特征是其在猫使用是禁忌顺铂已被证明是在猫23剧毒。如果患者有一个或只有几个结节，则建议使用肿瘤内路由。如果结节的数目超过〜5结节，则建议使用静脉内途径。顺铂的瘤内注射具有一定的抗肿瘤效果，即使没有电脉冲的应用，而博来霉素，由于其高的亲水性的性质，没有。因此，顺铂是首选的第一种药物，如果肿瘤内的应用是可能的犬。这是很重要的，因为这必须满足ECT的有效的抗肿瘤作用的第二个条件，那就是整个肿瘤具有足够高的电场的覆盖范围。以获得在细胞的增加膜透化，在组织内感应电场必须超过一定的阈值。在组织中的电场的分布取决于电极，因此可用于在什么样的电极是重要的。板或平行阵列电极可以安全地用于小浅表肿瘤24。如果肿瘤是更厚和更大的，六边形的电极更适合。一个重要的事实是，电脉冲的应用可以重复，以使整个肿瘤区域被覆盖，包括安全边际。因为电脉冲引起血管锁，它们的应用应当从肿瘤的边缘开始，朝它的中心。 目前，该技术的限制是其防止深层肿瘤的治疗电极的可用性。然而，电极的结构是一个技术特征是快速发展，现在不同组的电极用于˚F或特定施用长针单电极对结肠直肠肿瘤的转移瘤在肝脏25，骨转移7，软组织肉瘤26，大肠癌腔内的电极，胃和食管肿瘤27和膨胀&#39;伞&#39;型电极，用于脑瘤8 。 ECT可以有效地作为仅一次性处理，但在故障或局部肿瘤反应的情况下，它可以具有改善的效力可以重复几次。 ECT是一个局部消融治疗。其本地有效性达到局部肿瘤控制的80％，但没有对远处转移7，这是治疗的另一个限制明显的效果。因此，实现了全身反应，ECT可以与其他疗法结合。最近，我们提出用ECT电的另一个生物医学应用， 即免疫调节molec的基因电转移相结合ULE白细胞介素-12（IL-12），这是其抗肿瘤活性已知的。 ECT和IL-12基因电转移的组合已经被证明是无副作用和良好的肿瘤局部高完全缓解率28，这也阻止远处转移复发的一个安全的综合治疗。 总之，电化学的优点是它的简单性，治疗会话，低细胞抑制的剂量和微不足道的副作用的持续时间短。另外，它是在门诊基础上执行，因此，患者不必留在医院。 如今，在市场上的电脉冲的不同的发电机的可用性增加，还导致了发电机的价格与可靠性没有损失的降低，从而使电化学更加实惠。这些都应该导致兽药的广泛应用这一疗法。

ECT是对于不同的组织学类型的肿瘤的治疗的局部烧蚀技术。从药物诱导的细胞杀伤，这是在细胞中增加的药物积累的结果消融结果由于通过对药物的瘤内或静脉内注射后肿瘤周围皮肤直接递送电脉冲的诱导改变的细胞膜通透性。 ECT的主要机制是增强细胞抑制药物通过增加细胞内药物积累的有效性，通过细胞膜electropermeabilization（电）1,2。电:使两个细胞抑制剂的增高药的吸收:博来霉素，顺铂，这是亲水性药物膜通透性较差。由于博来霉素和顺铂的以电脉冲的应用位点的增加的细胞毒性，需要用于良好的抗肿瘤效力低的药物剂量，因此没有或最小的药物的全身性副作用是OBSERVED 3。 ECT的另一个特征是一种肿瘤抗血管作用这首先导致血管锁， 即 ，在一个血管破坏作用，降低肿瘤内的血流，并在其内的细胞抑制剂的结果保留和第二即 ，内皮细胞的直接杀伤小血管，导致继发性肿瘤细胞死亡4-6。 ECT的抗肿瘤效果的第三种机制是免疫系统的活化作为ECT诱发肿瘤抗原在肿瘤周围脱落的免疫应答。免疫系统的参与对于肿瘤7彻底根除至关重要。 ECT已在人类肿瘤，在那里，因为它具有最多的客观反应3 80％的ECT已作为标准治疗皮肤和皮下肿瘤显示出巨大的潜力。另外，ECT在若干临床研究中治疗肿瘤转移在大脑中使用<s达&gt; 8，9骨和肝脏10。在兽医肿瘤的临床研究已经表明在狗11,12，猫13和马14治疗皮肤和皮下肿瘤时ECT处理导致高达80％的长期目标的响应。的抗肿瘤效果取决于肿瘤的大小和肿瘤小于2 cm 3的响应比更大的肿瘤15更好。 此外，随着ECT在治疗狗肥大细胞瘤手术的比较研究表明，ECT治疗导致的完全反应70％，而手术导致了完全反应50％11证明，认为ECT是同等待遇手术。另外，ECT代表用于治疗实体肿瘤的外科治疗，特别是在这些情况下的另一种方法，当业主不同意手术，或者如果肿瘤是难以与边缘清晰大多是由于切除其位置（ 例如 ，浸润性大血管，头部或腿的肿瘤，在肛门区等 ）。本文的目的是要确定，以便安全地和适当地治疗狗和猫皮肤和皮下肿瘤的用于联合使用顺铂和博来霉素的电脉冲的施加ECT给予指令的操作程序。

<table><tbody><tr><td>Cisplatin Kobi</td><td>Fresenius Kobi Oncology, Hampshire, Great Britain</td><td></td><td>cytostatic drug</td></tr><tr><td>Bleomycin PCH</td><td>Pharmachemie B.V., Haarlem, Dutch</td><td></td><td>cytostatic drug</td></tr><tr><td>AquaUltra Basic</td><td>Ultragel, Budapest, Hungary</td><td></td><td>water based gel&amp;nbsp;</td></tr><tr><td>Electropulsator ELECTROvetS13&amp;nbsp;</td><td>Leroy biotech, Toulouse, France</td><td></td><td>square-wave pulse generator</td></tr></tbody></table>

operating procedures of the electrochemotherapy for treatment of tumor in dogs and cats

电化学（ECT）是一种用于治疗不同组织学的实体瘤的局部的方法。其作用机理是通过"电"的方式基于细胞的膜通透性。为了实现细胞的"电穿孔"，由发电机产生的电脉冲，并通过使用电极递送至靶组织。电穿孔是用于引进分子，如细胞生长抑制剂的药物，成不能靠自己细胞膜传递细胞的物理方法。在电化学，目前，顺铂和博来霉素在临床上使用。电化学抗肿瘤效果高，例如高达犬肥大细胞的100％的完全缓解肿瘤小于2 cm 3的达到了。此外，电化学可用于不能手术的肿瘤的治疗。之一的电化学的重要特征是，它可以有效地作为一个一次性治疗只要。然而，在故障或局部肿瘤反应的情况下，它可以具有相等或改进的效力重复几次。电化学已经在人类和兽医肿瘤学的各种组织学的皮肤和皮下肿瘤的标准治疗。此外，一些临床研究利用电化学的深层肿瘤持续。

案例展示位于狗的眼睑肥大细胞瘤。该患者用瘤内注射顺铂，并使用平板电极8电脉冲的应用处理。 肿瘤的大小为5mm。由于肿瘤的位置在全身麻醉的敏感区域使用。根据顺铂的肿瘤0.5毫升（0.5毫克）的大小瘤内注射。注射后一分钟，用8毫米的钢板电极施加的电脉冲。只有一个电脉冲的运行施加。治疗后病人被监督，直到它恢复意识足够。患者在两个小时后释放。一周后病人来，为要定期检查。肿瘤溃烂及肿瘤周围区域是微肿。肿瘤通过游标卡尺和拍照的测量。一个月后处理方法T中的肿瘤没有进一步的证据指出。缝合伤口，并与脱发瘢痕在治疗部位被看见。处理后的狗没有显示出疼痛的迹象，眼睑的功能已在整个愈合期保存和处理后的美容效果优异。 <img alt="图1" src="/files/ftp_upload/54760/54760fig1.jpg" /> 图1. 电化学疗法治疗。示出处理（A）前的狗的图中，一周（B）中 ，一个月（C）和三个月后处理（D）。在下眼睑上（A）肥大细胞瘤。 2.5岁的德国拳师狗与之前的治疗5mm的肿瘤大小。 （B）中的治疗后，肿瘤开始溃疡和围绕该电穿孔区域中的组织变得轻微肿胀。我们两个EKS治疗后形成的肿瘤部位痂掉了下来（未显示）。 （C）的电化学无肿瘤的证据指出，并与脱发瘢痕在肿瘤部位被视为后一个月。 （D）总电化学再上皮化后3个月内发生肿瘤的部位。 <a href="https://www.jove.com/files/ftp_upload/54760/54760fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td></td><td colspan="2">操作模式（OM） </td></tr><tr><td>数，体积和肿瘤结节的位置</td><td>一个</td><td>乙</td></tr><tr><td> 1 - 3结节&lt;1立方厘米皮肤及口腔肿瘤</td><td> √ </td><td></td></tr><tr><td> &gt; 3结节，任何音量皮肤及口腔肿瘤头</td><td></td><td> √ </tD&gt; </tr><tr><td>任何数量，皮肤及口腔肿瘤头的任何卷</td><td></td><td> √ </td></tr></tbody></table> 表1.工作模式的选择是基于肿瘤结节和它们的大小数深度镇静= OM一个。全身麻醉= OM B. <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>肿瘤化疗药物/尺寸</td><td>瘤内的应用程序</td><td>静脉应用</td></tr><tr><td>顺铂的剂量（1毫克/毫升）&lt;1 cm 3的</td><td> 1毫升（1毫克）/肿瘤的厘米3 </td><td></td></tr><tr><td>顺铂的剂量（1毫克/毫升）&gt; 1 cm 3的</td><td> 0.5-1毫升（0.5-1毫克）/肿瘤厘米3 </td><td></td></tr><tr><td>博莱霉素的剂量（3000国际单位/毫升）&lt;1 cm 3的</td><td> 0.5毫升（1500 IU）致瘤的/厘米3- [R </td><td> 300 IU /公斤</td></tr><tr><td>博莱霉素的剂量（3000国际单位/毫升）&gt; 1 cm 3的</td><td> 0.5 - 1毫升（1,500-3,000 IU）/肿瘤厘米3 </td><td> 300 IU /公斤</td></tr></tbody></table> 表2.根据犬给药途径和肿瘤大小的细胞抑制剂的适当的注入量。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>肿瘤化疗药物/尺寸</td><td>瘤内的应用程序</td><td>静脉应用</td></tr><tr><td>博莱霉素的剂量（3000国际单位/毫升）&lt;1 cm 3的</td><td> 0.5毫升（1500 IU）的肿瘤/ cm 3的</td><td> 300 IU /公斤</td></tr><tr><td>博莱霉素的剂量（3000国际单位/毫升）&gt; 1 cm 3的</td><td> 0.5 - 1毫升（1500 - 3000 IU）肿瘤/厘米3 </td><td> 300 IU /公斤</td></tr></tbody></table>表3.根据肿瘤大小在猫的细胞抑制剂的适当的注入量。

Watch this Scientific Journal Video about Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats at JoVE.com

Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

Electrochemotherapy is the local treatment of solid tumors. The main mechanism is electroporation-mediated permeabilization of the tumor cells' membrane which enables increased internalization of cytostatics like bleomycin and cisplatin. Thus, antitumor effectiveness is potentiated at the site of the application of electric pulses to the tumors.

在电化学疗法的操作程序在肿瘤治疗中的狗和猫

Electrochemotherapy (ECT) is a local approach which is used for treating solid tumors of different histologies. Its mechanism is based on cell membrane ...

operating-procedures-electrochemotherapy-for-treatment-tumor-dogs

Research

JoVE Journal

Cancer Research

26.4K 次观看.  University of Ljubljana. Electrochemotherapy is the local treatment of solid tumors. The main mechanism is electroporation-mediated permeabilization of the tumor cells' membrane which enables increased internalization of cytostatics like bleomycin and cisplatin. Thus, antitumor effectiveness is potentiated at the site of the application of electric pulses to the tumors. 

视频: Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation

Pancreatic cancer (PC), a disease which kills approximately 40,000 patients each year in the US, has successfully evaded several therapeutic approaches including the promising immunotherapeutic strategies. Irreversible electroporation (IRE) is a non-thermal ablation technique that induces tumor cell death without destruction of adjacent collagenous structures, thus enabling the procedure to be performed in tumors very close to blood vessels. Unlike thermal ablation techniques, IRE results in gradual apoptotic cell death, along with immediate ablation induced necrosis, and is currently in clinical use for selected patients with locally advanced PC. An ablative, non-target specific procedure like IRE can induce a myriad of responses in the tumor microenvironment. A few studies have addressed the effects of IRE on tumor growth in other tumor types, but none have focused on PC. We have developed a syngeneic mouse model of PC in which subcutaneous (SQ) and orthotopic tumors can be successfully treated with IRE in a highly controlled setting, facilitating various longitudinal studies post procedure. This animal model serves as a robust system to study the effects of IRE and ways to improve the clinical efficacy of IRE.

Irreversible electroporation (IRE) is a non-thermal ablation technique used for the treatment of locally advanced pancreatic cancer. Being a relatively new technique, the effects of IRE on the tumor growth are poorly understood. We have developed a syngeneic mouse model that facilitates studying the effects of IRE on pancreatic cancer.

自体胰腺癌小鼠模型研究不可逆电穿孔的影响

Pancreatic cancer (PC), a disease which kills approximately 40,000 patients each year in the US, has successfully evaded several therapeutic approaches ...

科研

癌症研究

One Minute, Sub-One-Watt Photothermal Tumor Ablation Using Porphysomes, Intrinsic Multifunctional Nanovesicles

We recently developed porphysomes as intrinsically multifunctional nanovesicles. A photosensitizer, pyropheophorbide &alpha;, was conjugated to a phospholipid and then self-assembled to liposome-like spherical vesicles. Due to the extremely high density of porphyrin in the porphyrin-lipid bilayer, porphysomes generated large extinction coefficients, structure-dependent fluorescence self-quenching, and excellent photothermal efficacy. In our formulation, porphysomes were synthesized using high pressure extrusion, and displayed a mean particle size around 120 nm. Twenty-four hr post-intravenous injection of porphysomes, the local temperature of the tumor increased from 30 &deg;C to 62 &deg;C rapidly upon one minute exposure of 750 mW (1.18 W/cm2), 671 nm laser irradiation. Following the complete thermal ablation of the tumor, eschars formed and healed within 2 weeks, while in the control groups the tumors continued to grow and all reached the defined end point within 3 weeks. These data show how porphysomes can be used as potent photothermal therapy (PTT) agents.

We developed novel intrinsic multifunctional nanovesicles called porphysomes, which have structure-dependent fluorescence self-quenching and unique photothermal properties, thus functioning as potent photothermal therapy agents. We formulated porphysomes using high pressure extrusion and investigated their photothermal therapy efficacy in a xenograft tumor model.

一分钟，小一瓦光热消融肿瘤使用Porphysomes，内在多功能纳米囊泡

We recently developed porphysomes as intrinsically multifunctional nanovesicles. A photosensitizer, pyropheophorbide &alpha;, was conjugated to a ...

生物工程

Electrochemotherapy of Tumours

Electrochemotherapy is a combined use of certain chemotherapeutic drugs and electric pulses applied to the treated tumour nodule. Local application of electric pulses to the tumour increases drug delivery into cells, specifically at the site of electric pulse application. Drug uptake by delivery of electric pulses is increased for only those chemotherapeutic drugs whose transport through the plasma membrane is impeded. Among many drugs that have been tested so far, bleomycin and cisplatin found their way from preclinical testing to clinical use. Clinical data collected within a number of clinical studies indicate that approximately 80% of the treated cutaneous and subcutaneous tumour nodules of different malignancies are in an objective response, from these, approximately 70% in complete response after a single application of electrochemotherapy. Usually only one treatment is needed, however, electrochemotherapy can be repeated several times every few weeks with equal effectiveness each time. The treatment results in an effective eradication of the treated nodules, with a good cosmetic effect without tissue scarring.

Electrochemotherapy is a combined use of certain chemotherapeutic drugs such as cisplatin and bleomycin and electric pulses applied to the treated tumour nodule. We present the clinical protocol of electrochemotherapy for treatmen of subcutaneous and cutaneous metastases of melanoma.

电化学的肿瘤

Electrochemotherapy is a combined use of certain chemotherapeutic drugs and electric pulses applied to the treated tumour nodule. Local application of ...

生物学

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Flexible Organic Electronic Devices for Pulsed Electric Field Therapy of Glioblastoma

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed electric fields (PEFs) are promising for the improvement of treatment efficiency. However, they rely on rigid electrodes that cause acute and chronic damage, especially in soft tissues such as the brain. In this work, flexible electronics were used to deliver PEFs to tumors and the biological response was evaluated with fluorescent microscopy. Interdigitated gold electrodes on a thin, transparent parylene-C substrate were coated with the conducting polymer PEDOT:PSS, resulting in a conformable and biocompatible device. The effects of PEFs on tumors and their microenvironment were examined using various biological models. First, monolayers of glioblastoma cells were cultured on top of the electrodes to investigate phenomena in vitro. As an intermediate step, an in ovo model was developed where engineered tumor spheroids were grafted in the embryonic membrane of a quail. Due to the absence of an immune system, this led to highly vascularized tumors. At this early stage of development, embryos have no immune system, and tumors are not recognized as foreign bodies. Thus, they can develop fast while developing their own vessels from the existing embryo vascular system, which represents a valuable 3D cancer model. Finally, flexible electrode delivery of PEFs was evaluated in a complete organism with a functional immune system, using a syngenic, orthograft (intracranial) mouse model. Tumor spheroids were grafted into the brain of transgenic multi-fluorescent mice prior to the implantation of flexible organic electrode devices. A sealed cranial window enabled multiphoton imaging of the tumor and its microenvironment during treatment with PEFs over a period of several weeks.

This work describes the development of flexible interdigitated electrodes for implementation in 3D brain tumor models, namely, in vitro culture, in ovo model, and in vivo murine model. The proposed method can be used to evaluate the effects of pulsed electric fields on tumors at different levels of complexity.

用于胶质母细胞瘤脉冲电场治疗的柔性有机电子器件

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed ...

Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells In Vitro

In recent years, there has been the difficulty in finding more effective therapies against cancer with less systemic side effects. Therefore Photodynamic Therapy is a novel approach for a more tumor selective treatment.
Photodynamic Therapy (PDT) that makes use of a nontoxic photosensitizer (PS), which, upon activation with light of a specific wavelength in the presence of oxygen, generates oxygen radicals that elicit a cytotoxic response1. Despite its approval almost twenty years ago by the FDA, PDT is nowadays only used to treat a limited number of cancer types (skin, bladder) and nononcological diseases (psoriasis, actinic keratosis)2.
The major advantage of the use of PDT is the ability to perform a local treatment, which prevents systemic side effects. Moreover, it allows the treatment of tumors at delicate sites (e.g. around nerves or blood vessels). Here, an intraoperative application of PDT is considered in osteosarcoma (OS), a tumor of the bone, to target primary tumor satellites left behind in tumor surrounding tissue after surgical tumor resection. The treatment aims at decreasing the number of recurrences and at reducing the risk for (postoperative) metastasis.
In the present study, we present in vitro PDT procedures to establish the optimal PDT settings for effective treatment of widely used OS cell lines that are used to reproduce the human disease in well established intratibial OS mouse models. The uptake of the PS mTHPC was examined with a spectrophotometer and phototoxicity was provoked with laser light excitation of mTHPC at 652 nm to induce cell death assessed with a WST-1 assay and by the counting of surviving cells. The established techniques enable us to define the optimal PDT settings for future studies in animal models. They are an easy and quick tool for the evaluation of the efficacy of PDT in vitro before an application in vivo.

Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells In Vitro

The protocol reported here allows the evaluation of the efficacy of photodynamic therapy (PDT) in cell lines and the optimization of the PDT settings before applying the therapy in animal models.

光动力疗法治疗骨肉瘤细胞的细胞毒功效<em&gt;体外</em

In recent years, there has been the difficulty in finding more effective therapies against cancer with less systemic side effects. Therefore Photodynamic ...

医学

通过基于电穿孔的质粒 DNA 递送进行体内脑肿瘤建模

Modeling Brain Tumors In Vivo Using Electroporation-Based Delivery of Plasmid DNA Representing Patient Mutation Signatures

Tumor models are critical for the preclinical testing of brain tumors in terms of exploring new, more efficacious treatments. With significant interest in immunotherapy, it is even more critical to have a consistent, clinically pertinent, immunocompetent mouse model to examine the tumor and immune cell populations in the brain and their response to treatment. While most preclinical models utilize orthotopic transplantation of established tumor cell lines, the modeling system presented here allows for a "personalized" representation of patient-specific tumor mutations in a gradual, yet effective development from DNA constructs inserted into dividing neural precursor cells (NPCs) in vivo. DNA constructs feature the mosaic analysis with the dual-recombinase-mediated cassette exchange (MADR) method, allowing for single-copy, somatic mutagenesis of driver mutations. Using newborn mouse pups between birth and 3 days old, NPCs are targeted by taking advantage of these dividing cells lining the lateral ventricles. Microinjection of DNA plasmids (e.g., MADR-derived, transposons, CRISPR-directed sgRNA) into the ventricles is followed by electroporation using paddles that surround the rostral region of the head. Upon electrical stimulation, the DNA is taken up into the dividing cells, with the potential of integrating into the genome. The use of this method has successfully been demonstrated in developing both pediatric and adult brain tumors, including the most common malignant brain tumor, glioblastoma. This article discusses and demonstrates the different steps of developing a brain tumor model using this technique, including the procedure of anesthetizing young mouse pups, to microinjection of the plasmid mix, followed by electroporation. With this autochthonous, immunocompetent mouse model, researchers will have the ability to expand preclinical modeling approaches, in efforts to improve and examine efficacious cancer treatment.

作者聚焦:使用基于电穿孔的质粒 DNA 递送代表患者突变特征的体内脑肿瘤建模  

Utilizing an immunocompetent, autochthonous tumor model driven by common patient mutations for preclinical testing is critical for immunotherapeutic testing. This protocol describes a method to generate brain tumor mouse models using electroporation-based delivery of plasmid DNA that represent common patient mutations, thus providing an accurate, reproducible, and consistent mouse model.

使用基于电穿孔的质粒DNA递送对 体内 脑肿瘤进行建模，以代表患者突变特征

Tumor models are critical for the preclinical testing of brain tumors in terms of exploring new, more efficacious treatments. With significant interest in ...

In Vitro and In Vivo Evaluation of Photocontrolled Biologically Active Compounds - Potential Drug Candidates for Cancer Photopharmacology

Photocontrolled, biologically active compounds are an emerging class of &#34;smart&#34; drug candidates. They provide additional safety in systemic chemotherapy due to their precise spatiotemporal activation by directing a benign, non-ionizable light to a specific location within the patient&#39;s body. This paper presents a set of methods to evaluate the in vitro potency and ex vivo efficiency of the photoactivation of photocontrolled, biologically active compounds as well as the in vivo efficacy at early stages of drug development. The methodology is applied to anticancer cytotoxic peptides, namely, the diarylethene-containing analogs of a known antibiotic, gramicidin S. The experiments are performed using 2D (adherent cells) and 3D (spheroids) cell cultures of a cancer cell line (Lewis lung carcinoma, LLC), live tissue surrogates (pork meat mince), and an allograft cancer model (subcutaneous LLC) in immunocompetent mice. The selection of the most effective compounds and estimation of realistic phototherapeutic windows are performed via automated fluorescence microscopy. The photoactivation efficiency at varying illumination regimens is determined at different depths in a model tissue, and the optimal light dosage is applied in the final therapeutic in vivo experiment.

In Vitro and In Vivo Evaluation of Photocontrolled Biologically Active Compounds - Potential Drug Candidates for Cancer Photopharmacology

This protocol presents a set of experiments adopted for the evaluation of photoswitchable anticancer peptides, that can be used in the preclinical screening of such compounds. This includes cytotoxicity assessment in 2D and 3D cell cultures, the evaluation of ex vivo (model tissue) photoisomerization efficiency, and in vivo efficacy.

光控生物活性化合物的体外和体内评价 - 癌症光药理学的潜在候选药物

Photocontrolled, biologically active compounds are an emerging class of &#34;smart&#34; drug candidates. They provide additional safety in systemic ...

在电化学疗法的操作程序在肿瘤治疗中的狗和猫

摘要

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此视频中的章节

自体胰腺癌小鼠模型研究不可逆电穿孔的影响

一分钟，小一瓦光热消融肿瘤使用Porphysomes，内在多功能纳米囊泡

电化学的肿瘤

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

用于胶质母细胞瘤脉冲电场治疗的柔性有机电子器件

光动力疗法治疗骨肉瘤细胞的细胞毒功效

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征

光控生物活性化合物的体外和体内评价 - 癌症光药理学的潜在候选药物

在电化学疗法的操作程序在肿瘤治疗中的狗和猫

摘要

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此视频中的章节

自体胰腺癌小鼠模型研究不可逆电穿孔的影响

一分钟，小一瓦光热消融肿瘤使用Porphysomes，内在多功能纳米囊泡

电化学的肿瘤

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

用于胶质母细胞瘤脉冲电场治疗的柔性有机电子器件

光动力疗法治疗骨肉瘤细胞的细胞毒功效

使用基于电穿孔的质粒DNA递送对 体内 脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对 体内 脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对 体内 脑肿瘤进行建模，以代表患者突变特征

光控生物活性化合物的体外和体内评价 - 癌症光药理学的潜在候选药物

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征

使用基于电穿孔的质粒DNA递送对体内脑肿瘤进行建模，以代表患者突变特征