支持这项工作是由德意志研究联合会（SFB 656，明斯特，德国，C7和C6项目）和IZKF的明斯特（核心单元SMAP）。作者感谢特鲁茨范乐，安妮Kanzog，乌特纽格伯尔，Wiebke Gottschlich和罗马PRIEBE的优良的技术援助和生产放射性示踪剂丹尼尔布尔克特和Sven Fatum。

1。捐赠者的器官恢复<ol><li>立体显微镜，记录重量8-10周龄大鼠（捐赠者和接受者体重应该匹配）。 </li><li>麻醉供体鼠（刘易斯·布朗挪威F1，F1 LBN）使用氧气/异氟醚吸入异氟醚4％/ 2升/分钟氧气。 2-2.5％降低异氟醚维持麻醉。 </li><li>将大鼠麻醉手术垫。修复大鼠的四肢用胶带将垫和申请眼科的软膏（Bepanthen，拜耳）大鼠的眼睛。 </li><li>刮脸和大鼠腹部消毒，并执行下腹正中切口从耻骨尾边境的肝脏。通过仔细向右侧移动肠道暴露左肾和其船舶。放置在左肾纱布薄片，湿润温热的等渗盐溶液，以防止干燥。 </li><li>仔细剖析脂肪组织与杜蒙弯角尖镊子（杜蒙SS-Inox公司45镊子医疗，精细的科学工具CAT.NO. 11203-25）包围左侧输尿管。避免输尿管的任何直接接触，而不是有足够的周围的脂肪组织，调动。 </li><li>使用杜蒙弯角尖镊子分离左肾静脉肾动脉。从两艘船卸下所有的脂肪和结缔组织，烧灼肾上腺和睾丸血管。 </li><li>左肾动脉和静脉解剖主动脉和下腔静脉（IVC）的上方和下方交界处，分别为，灸所有传出的动脉。 </li><li>夹住主动脉，左肾动脉显微钳（微Serrefines，精细的科学工具18055-04 CAT.NO.）以上，尽可能靠近肠系膜上动脉。结扎腹主动脉使用手术丝绸（5-0，Vömel，艺术，什么14739）左肾动脉约5毫米以下。 </li><li>结扎手术丝绸下腹IVC（5-0Vömel）和钳位肾上腺IVC显微钳（的微Serrefines，精细的科学工具18055-03 CAT.NO.）。 </li><li>切尽可能靠近下腔静脉肾静脉用细剪刀（虹膜剪 - ToughCut直11.5厘米，精细的科学工具货号14058-11）。 </li><li>慢慢灌注到肾脏原位用2毫升冰冷HTK的灌注溶液（HTK CUSTODIOL，Franz博士科勒化学），通过插入一个插管（Microlance 3 27G¾，BD，货号302200）在腹主动脉。确认肾的颜色变化（现在橄榄有色）。 </li><li>首先，横切主动脉，然后下腹主动脉，最后尽可能接近膀胱输尿管。 </li><li>连同输尿管和存储HTK灌注液冰取出肾脏和血管供应。 </li><li>安乐死捐助大鼠通过消除麻醉下的心脏血管钳和连续切除。 </li></ol> 2。收件人的制备及移植重刑<ol><li>设置的立体显微镜，记录重量的大鼠。 </li><li>麻醉收件人使用异氟醚4％的大鼠（刘易斯）。 2-2.5％降低异氟醚维持麻醉。 </li><li>将大鼠麻醉手术垫。修复大鼠的四肢用胶带将垫和眼药膏老鼠`眼睛。监视和控制核心体温，使用的直肠温度传感器和变暖垫。在手术过程中重复地控制温度，呼吸和心脏的汇率的动物。 </li><li>刮脸和消毒腹部和执行下腹正中切口从耻骨尾边境的肝脏。使用手术刀forcutting的皮肤，以尽量减少皮肤损伤。暴露左肾和其货船的向右侧移动肠道。 </li><li>小心取出肾胶囊使用杜蒙弯角尖镊子和棉签。 </li><li>闭塞左侧肾动脉，静脉和输尿管附近的肾肺门结扎手术线（4-0 MERSILENE的，爱惜康的猫。EH6732H）。海关大部份的肾脏，只留下肾门。用棉签清洁切割表面，并检查是否有出血。如有必要添加另一个结扎。 </li><li>仔细解剖直言不讳地通过结缔组织的覆盖尾现场用棉签上的腹主动脉和下腔静脉。 </li><li>从腹主动脉和下腔静脉使用杜蒙弯角尖镊子分离任何可见的神经。 </li><li>小心解剖，通过下腹主动脉和腔静脉之间的结缔组织片，形成两个开口2-4毫米长:第一个下面的睾丸血管分支，而第二个以上的分支髂总血管。烧灼髂腰动脉和静脉之间。 </li><li>绘制一个单一的手术线（MERSILENE 0爱惜，的猫。no.EH6665E），通过两个开口。使用这些线程拉下腹主动脉和腔静脉轻轻地动物获得背站点上的血管分支的右侧。烧灼两个开口之间的任何这些船只，随后移除手术线程。 </li><li>接下来，主动脉和下腔静脉闭塞顺时针应用四个显微外科的夹具（微Serrefines，精细的科学工具18055-04 CAT.NO.）开始在上开口（微Serrefines的腔静脉上部开口与主动脉和结束精细的科学工具18055-03 CAT.NO.）。 </li><li>后来仔细刺破上结束孤立部分的主动脉用针（Microlance 3 27G¾，BD，货号302200），插入刀片罚款弹簧剪刀（维纳斯弹簧剪刀 - 3毫米的刀片，精细科学工具，15000-00 cat.no），通过穿刺口，并进行很短的纵形切口。用冰冷的HTK的灌注液，以除去任何剩余的血液冲洗大鼠主动脉。 </li><li>同样地，小心地捅破较低的结局的隔离部分的腔静脉用针，通过穿刺开口插入下部的细弹簧剪刀刀片和下方的主动脉切口不久结束执行中形成纵向切口。再次，刷新船只，以消除任何剩余的血液。 </li><li>取出移植肾的存储解决方案。将低于前左肾位置，并确保它是正确的导向。 </li><li>检查移植肾动脉的方向，并确保不存在任何曲折。一层薄薄的纱布，然后将移植和滋润它用冰冷的HTK灌注液冷却，防止干燥。 </li><li>注视开幕肾上腺主动脉片的移植肾动​​脉通向手术线（9-0 Ethilon，爱惜康猫2809G） - 首先在上，然后在低端主动脉切口。顺时针用手术线连续缝合关闭切口（9-0 Ethilon），杜蒙弯角尖镊子和弯曲卡斯特罗维霍持针器（精细的科学工具的猫。12061-01）。之后，通过一个逆时针的第二个连续缝合覆盖第一结缔组织缝合。现在，只有移植物的主动脉片的下部被关闭。定期用冰冷的HTK灌注液冷却移植。 </li><li>灌注移植物用1ml冰冷的HTK灌注液通过验证缝线的气密性和完整性，以及除去其它的血液在接枝移植物的主动脉片的开口端插入的钝针。之后，结扎主动脉手术丝绸片（5-0Vömel）的开口端。 </li><li>检查移植肾静脉的方向，并确保有没有曲折。 </li><li>固定的静脉移植手术线程（9-0 Ethilon） - 首先在上部的开口，然后在其下端的切口在腔静脉。关闭切口顺时针连续缝合，用手术线（Ethilon 9-0），杜蒙弯角尖镊子和弯曲卡斯特罗维霍持针器。定期用冰冷的HTK灌注液冷却移植。 </li><li>逆时针删除显微夹，开始与最腔静脉钳。移除最后主动脉钳后，小心地用棉签最终轻度出血停止。几秒钟后，移植将转红润有色，和uretric宫缩应该出现。 </li><li>对于插入输尿管移植到收件人`膀胱小心地取出一小块肌肉，在膀胱顶部的使用维纳斯弹簧剪刀（学生维纳斯直弹簧剪刀，精细的科学工具，货号91500-09 ）。然后，除去周围组织（主要是脂肪）移植输尿管杜蒙弯角尖镊子。因此，仔细抢输尿管尖端，轻轻分开周围的脂肪，结缔组织和嵌入式船只来回米，通过在相反的方向拉出。固定uretric的尖的手术线的端部（6-0聚丙烯缝线，Ethicon的，猫8697H）穿孔预先制备的膀胱顶部与附加的针。拉输尿管膀胱和退出在该基地通过第二穿孔。 </li><li>先前固定在顶部用手术线（9-0 Ethilon）的膀胱输尿管分离的组织。切断的输尿管与附加的手术线的前端，拉从下面轻轻推入膀胱输尿管。之后，关闭用手术线（9-0 Ethilon），第二膀胱穿刺。 </li><li>关闭腹壁和皮肤两个连续的，但独立的缝合线，用手术缝合线（MERSILENE 0）。消毒伤口使用碘Pividon和管理丁丙诺啡（0.1毫克/每公斤体重/投标）SC三天手术后伤口疼痛控制。 </li></ol> 3。成像移植排斥反应<ol><li>非空腹大鼠麻醉使用异氟醚2-2.5％。 </li><li> 30活度FDG（30活度FDG在0.1毫升0.9％的氯化钠）静脉给药通过一个尾静脉置管使用24 G导管（布劳恩Introcan 4252500-01）。之后，用0.9毫升0.9％NaCl溶液中清除导管。 </li><li>让大鼠在一个限制器，无需麻醉，直到开始扫描和水合物动物静脉注射1毫升0.9％NaCl溶液每小时。 </li><li>重新麻醉大鼠扫描开始前立即使用异氟醚2％。 </li><li>将麻醉大鼠在高分辨率多丝室为基础的动物PET:相机quadHIDAC（牛津正电子系统有限公司，牛津大学，英国）。 PET扫描仪的空间分辨率为1.0毫米，并在整个视场角（直径，165毫米，轴向长度280毫米）是恒定的。在扫描过程中使用直肠温度传感器和控制麻醉与脉冲血氧监测和控制体温。 </li><li>启动动态采集为60米FDG注射开始后180分钟减少示踪剂在肾脏FDG经肾脏排泄造成的积累。 </li><li>应用5 18 F-氟化四活度，并执行另一个PET扫描后立即18 F-氟注射60分钟，而在扫描仪中的大鼠肾实质的识别和计算的18 F-间隙5的位置移动。从两个扫描重建图像。手动跟踪重建图像上感兴趣体积（VOI）2分钟后，18架F-氟注射液（灌注相）周围肾皮质FDG图像传输VOI。仔细排除肾盂从VOI。重建图像扫描和手动跟踪周围肾皮质感兴趣体积（VOI）。列表模式数据重建成图像的一个像素大小为0.4×0.4×0.4毫米3。仔细排除肾盂从VOI。 </li><li>计算FDG摄取比O比数的计数和体积，并计算出注射剂量的百分比（％ID）。我们使用MATLAB和断层成像（TIM）2.8版（R2011b版，MathWorks公司，Natic​​k市，MA，USA）进行图像分析。 </li></ol>

<ol><li>Wu, O., Levy, A. R., Briggs, A., Lewis, G., Jardine, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Acute+rejection+and+chronic+nephropathy%3A+a+systematic+review+of+the+literature%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">Acute rejection and chronic nephropathy: a systematic review of the literature.</a> Transplantation. 87, 1330-1339 (2009).</li>
<li>Miller, B. F., Gonzales, E., Wilchins, L. J., Nathan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Kidney+transplantation+in+the+rat%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Kidney transplantation in the rat.</a> Nature. 194, 309-310 (1962).</li>
<li>Reuter, S., Schnöckel, U., Schröter, R., Schober, O., Pavenstädt, H., Schäfers, M., Gabriëls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Non-invasive+imaging+of+acute+renal+allograft+rejection+in+rats+using+small+animal+F-FDG-PET%5BTitle%5D)+AND+%22PLoS.+One%22%5BJournal%5D)">Non-invasive imaging of acute renal allograft rejection in rats using small animal F-FDG-PET.</a> PLoS. One. 4, e5296(2009).</li>
<li>Reuter, S., Schnöckel, U., Edemir, B., Schröter, R., Kentrup, D., Pavenstädt, H., Schober, O., Schlatter, E., Gabriëls, G., Schäfers, M. Potential of noninvasive serial assessment of acute renal allograft rejection by 18F-FDG PET to monitor treatment efficiency. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aReuter%2C+S+%22J.+Nucl.+Med%22">J. Nucl. Med</a>. 51, 1644-1652 (2010).</li>
<li>Schnöckel, U., Reuter, S., Stegger, L., Schlatter, E., Schäfers, K. P., Hermann, S., Schober, O., Gabriëls, G., Schäfers, M. Dynamic 18F-fluoride small animal PET to noninvasively assess renal function in rats. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aSchn%C3%B6ckel%2C+U+%22Eur.+J.+Nucl.+Med.+Mol.+Imaging%22">Eur. J. Nucl. Med. Mol. Imaging</a>. 35, 2267-2274 (2008).</li>
<li>Grabner, A., Schnöckel, U., Kentrup, D., Schäfers, M., Reuter, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Strategies+for+Non-Invasive+Molecular+Imaging+of+Acute+Allograft+Rejection+by+Gamma+Scintigraphy+and+Positron+Emission+Tomography%5BTitle%5D)+AND+%22Curr.+Radiopharm%22%5BJournal%5D)">Strategies for Non-Invasive Molecular Imaging of Acute Allograft Rejection by Gamma Scintigraphy and Positron Emission Tomography.</a> Curr. Radiopharm. 4, 10-23 (2011).</li>
<li>Racusen, L. C., Solez, K., Colvin, R. B., Bonsib, S. M., Castro, M. C., Cavallo, T., Croker, B. P., Demetris, A. J., Drachenberg, C. B., Fogo, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+Banff+97+working+classification+of+renal+allograft+pathology%5BTitle%5D)+AND+%22Kidney+Int%22%5BJournal%5D)">The Banff 97 working classification of renal allograft pathology.</a> Kidney Int. 55, 713-723 (1999).</li>
<li>Edemir, B., Reuter, S., Borgulya, R., Schröter, R., Neugebauer, U., Gabriëls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Acute+rejection+modulates+gene+expression+in+the+collecting+duct%5BTitle%5D)+AND+%22J.+Am.+Soc.+Nephrol%22%5BJournal%5D)">Acute rejection modulates gene expression in the collecting duct.</a> J. Am. Soc. Nephrol. 19, 538-546 (2008).</li>
<li>Velic, A., Gabriëls, G., Hirsch, J. R., Schröter, R., Edemir, B., Paasche, S., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Acute+rejection+after+rat+renal+transplantation+leads+to+downregulation+of+Na%2B+and+water+channels+in+the+collecting+duct%5BTitle%5D)+AND+%22Am.+J.+Transplant%22%5BJournal%5D)">Acute rejection after rat renal transplantation leads to downregulation of Na+ and water channels in the collecting duct.</a> Am. J. Transplant. 5, 1276-1285 (2005).</li>
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</ol>

没有利益冲突的声明。

FDG-PET成像急性排斥反应的诊断是一个新的选择。由于其非侵入性和具体性，FDG-PET是古典针芯活检诊断比较优势。在一个切片检查，FDG-PET分析整个移植到有限的样本大小。此外，人们可以将它应用抗凝治疗的患者，可以进行重复例如 PET措施来监测治疗效率4。此外，我们已经表明两个主要可以区分开的鉴别诊断，即急性肾小管坏死引起的缺血再灌注损伤和急性calcineurin抑制剂毒性，...

<table><tbody><tr><td colspan="4">&lt;strong&gt;Equipment&lt;/strong&gt;</td></tr><tr><td>Mathieu Needle Holder - 14 cm</td><td>Fine Science Tools</td><td>12010-14</td><td></td></tr><tr><td>Castroviejo Micro Needle Holder - 9 cm</td><td>Fine Science Tools</td><td>12061-01</td><td></td></tr><tr><td>Surgical Scissors - Sharp_Blunt</td><td>Fine Science Tools</td><td>14001-12</td><td></td></tr><tr><td>Iris Scissors - ToughCut Straight 11.5 cm</td><td>Fine Science Tools</td><td>14058-11</td><td></td></tr><tr><td>Student Vannas Spring Scissors </td><td>Fine Science Tools</td><td>91500-09</td><td></td></tr><tr><td>Vannas Spring Scissors - 3 mm Blades</td><td>Fine Science Tools</td><td>15000-00</td><td></td></tr><tr><td>Student Tissue Forceps - 1x2 Teeth 12 cm</td><td>Fine Science Tools</td><td>91121-12</td><td></td></tr><tr><td>Dumont SS-45 Forceps - Inox Medical</td><td>Fine Science Tools</td><td>11203-25</td><td></td></tr><tr><td>Micro-Serrefine Clip Applicator with Lock</td><td>Fine Science Tools</td><td>18056-14</td><td></td></tr><tr><td>Micro-Serrefine 6 mm x 1 mm</td><td>Fine Science Tools</td><td>18055-03</td><td></td></tr><tr><td>Micro-Serrefine 4 mm x 0.75 mm</td><td>Fine Science Tools</td><td>18055-04</td><td></td></tr><tr><td colspan="4">&lt;strong&gt;Reagent&lt;/strong&gt;</td></tr><tr><td>Isoflurane (&lt;em&gt;e.g.&lt;/em&gt; Forene 100% v/v)</td><td>Abott</td><td></td><td></td></tr><tr><td>cutane antiseptic (&lt;em&gt;e.g.&lt;/em&gt; Octeniderm) </td><td>Sch&amp;#252;lke</td><td></td><td></td></tr><tr><td>Povidone Iodine (&lt;em&gt;e.g.&lt;/em&gt; Betaisodona)</td><td>Mundipharma</td><td></td><td></td></tr><tr><td>ophthalmic ointment (&lt;em&gt;e.g.&lt;/em&gt; Bepanthen)</td><td>Bayer</td><td></td><td></td></tr><tr><td>Buprenorphin (&lt;em&gt;e.g.&lt;/em&gt; Temgesic)</td><td>RB Pharmaceuticals</td><td></td><td></td></tr><tr><td>HTK perfusion solution (&lt;em&gt;e.g.&lt;/em&gt; CUSTODIOL HTK)</td><td>Dr. Franz K&amp;#246;hler Chemie</td><td></td><td></td></tr><tr><td>surgical thread Mersilene 0</td><td>Ethicon</td><td>EH6665E</td><td></td></tr><tr><td>surgical thread Mersilene 4-0</td><td>Ethicon</td><td>EH6732H</td><td></td></tr><tr><td>surgical thread Prolene 6-0</td><td>Ethicon</td><td>8697H</td><td></td></tr><tr><td>surgical thread Ethilon 9-0</td><td>Ethicon</td><td>2809G</td><td></td></tr><tr><td>surgical silk 5-0</td><td>V&amp;#246;mel</td><td>14739</td><td></td></tr><tr><td>Canula (&lt;em&gt;e.g.&lt;/em&gt; Microlance 3, 27G &amp;#190;)</td><td>BD</td><td>302200</td><td></td></tr></tbody></table>

non-invasive imaging of acute allograft rejection after rat renal transplantation using 18f-fdg pet

终末期肾脏疾病患者，肾移植的数量不断增加。发作的急性细胞移植排斥反应（AR）是一种消极的长期移植存活的预后因素，及时诊断是至关重要的移植肾功能1。目前，AR只能被确诊核心穿刺活检，其中，作为一种侵入性的方法，移植伤害甚至丧失露出显着的风险。此外，活检是不可行的患者服用抗凝药物和有限的采样点，这种技术可能会导致假阴性结果，当AR局灶性或片状。作为一个结果，这产生了一个持​​续的搜索新的AR的检测方法，包括使用各种移植模型的动物中，经常有许多工作要做。 由于60年代初大鼠肾移植是一种行之有效的实验方法为考试系统AR 2的振动性和分析。在此，我们除了小动物正电子发射断层扫描（PET）使用18架F-脱氧葡萄糖（FDG）评估AR单侧肾切除大鼠同种异体肾移植模型，并提出移植FDG-PET成像作为一种非侵入性的，具体的新选项和早期诊断的AR 3人的情况。此外，这种方法可应用于后续提高移植排斥反应的监测，4。

组织学 即主要是T淋巴细胞在AR白细胞，被招募到移植的排斥反应的严重程度，而被反射炎症的程度。在周期酸希夫染色（PAS）这里描绘（ 图1），肾移植显示显着的病理体征，即肾小球肾炎，小管炎，内皮炎和移植物浸润（ 图1，ATX POD4）（POD =术后一天AR）而拒绝的迹象都没有在本地控制肾（ 图1，CTR），移植物浸润细胞是高度消耗?...

Watch this Scientific Journal Video about Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET at JoVE.com

使用 18F-FDG PET 对大鼠肾移植后急性同种异体移植排斥进行无创成像医学|视频

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

在此，我们提出了一个大鼠肾移植模型，以非侵入性评估急性排斥反应，用18F-脱氧葡萄糖正电子发射断层扫描。

大鼠肾移植后急性排斥反应的非侵入性的成像使用<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

non-invasive-imaging-acute-allograft-rejection-after-rat-renal

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Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

Human Biology

Osmoregulation and Excretion

SCIENTISTS IN ACTION

16.9K 次观看.  University of Münster. 在此，我们提出了一个大鼠肾移植模型，以非侵入性评估急性排斥反应，用18F-脱氧葡萄糖正电子发射断层扫描。

视频: Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without the ability to target a specific or well-delineated tumor volume. The delivery of radiation was achieved using fixed radiation sources or linear accelerators producing megavoltage (MV) X-rays. These devices are unable to achieve sub-millimeter precision required for small animals. Furthermore, the high doses delivered to healthy surrounding tissue hamper response assessment. To increase the translation between small animal studies and humans, our goal was to mimic the treatment of human glioblastoma in a rat model. To enable a more accurate irradiation in a preclinical setting, recently, precision image-guided small animal radiation research platforms were developed. Similar to human planning systems, treatment planning on these micro-irradiators is based on computed tomography (CT). However, low soft-tissue contrast on CT makes it very challenging to localize targets in certain tissues, such as the brain. Therefore, incorporating magnetic resonance imaging (MRI), which has excellent soft-tissue contrast compared to CT, would enable a more precise delineation of the target for irradiation. In the last decade also biological imaging techniques, such as positron emission tomography (PET) gained interest for radiation therapy treatment guidance. PET enables the visualization of e.g., glucose consumption, amino-acid transport, or hypoxia, present in the tumor. Targeting those highly proliferative or radio-resistant parts of the tumor with a higher dose could give a survival benefit. This hypothesis led to the introduction of the biological tumor volume (BTV), besides the conventional gross target volume (GTV), clinical target volume (CTV), and planned target volume (PTV).
At the preclinical imaging lab of Ghent University, a micro-irradiator, a small animal PET, and a 7 T small animal MRI are available. The goal was to incorporate MRI-guided irradiation and PET-guided sub-volume boosting in a glioblastoma rat model.

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

PET 与 MRI 引导下微器对大鼠胶质瘤模型的照射

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without ...

科研

癌症研究

Multimodal Bioluminescent and Positronic-emission Tomography/Computational Tomography Imaging of Multiple Myeloma Bone Marrow Xenografts in NOG Mice

Multiple myeloma (MM) tumors engraft in the bone marrow (BM) and their survival and progression are dependent upon complex molecular and cellular interactions that exist within this microenvironment. Yet the BM microenvironment cannot be easily replicated in vitro, which potentially limits the physiologic relevance of many in vitro and ex vivo experimental models. These issues can be overcome by utilizing a xenograft model in which luciferase (LUC)-transfected 8226 MM cells will specifically engraft in the mouse skeleton. When these mice are given the appropriate substrate, D-luciferin, the effects of therapy on tumor growth and survival can be analyzed by measuring changes in the bioluminescent images (BLI) produced by the tumors in vivo. This BLI data combined with positronic-emission tomography/computational tomography (PET/CT) analysis using the metabolic marker 2-deoxy-2-(18F)fluoro-D-glucose (18F-FDG) is used to monitor changes in tumor metabolism over time. These imaging platforms allow for multiple noninvasive measurements within the tumor/BM microenvironment.

Here we use bioluminescent, X-ray, and positron-emission tomography/computed tomography imaging to study how inhibiting mTOR activity impacts bone marrow-engrafted myeloma tumors in a xenograft model. This allows for physiologically relevant, non-invasive, and multimodal analyses of the anti-myeloma effect of therapies targeting bone marrow-engrafted myeloma tumors in vivo.

nog 小鼠多发性骨髓瘤异种移植的多模态生物发光和正电子发射成形断层扫描成像

Multiple myeloma (MM) tumors engraft in the bone marrow (BM) and their survival and progression are dependent upon complex molecular and cellular ...

Dynamic Imaging of Chimeric Antigen Receptor T Cells with [18F]Tetrafluoroborate Positron Emission Tomography/Computed Tomography

T cells genetically engineered to express chimeric antigen receptors (CAR) have shown unprecedented results in pivotal clinical trials for patients with B cell malignancies or multiple myeloma (MM). However, numerous obstacles limit the efficacy and prohibit the widespread use of CAR T cell therapies due to poor trafficking and infiltration into tumor sites as well as lack of persistence in vivo. Moreover, life-threatening toxicities, such as cytokine release syndrome or neurotoxicity, are major concerns. Efficient and sensitive imaging and tracking of CAR T cells enables the evaluation of T cell trafficking, expansion, and in vivo characterization and allows the development of strategies to overcome the current limitations of CAR T cell therapy. This paper describes the methodology for incorporating the sodium iodide symporter (NIS) in CAR T cells and for CAR T cell imaging using [18F]tetrafluoroborate-positron emission tomography ([18F]TFB-PET) in preclinical models. The methods described in this protocol can be applied to other CAR constructs and target genes in addition to the ones used for this study.

Dynamic Imaging of Chimeric Antigen Receptor T Cells with [18F]Tetrafluoroborate Positron Emission Tomography/Computed Tomography

This protocol describes the methodology for non-invasively tracking T cells genetically engineered to express chimeric antigen receptors in vivo with a clinically available platform.

使用[18F]四氟硼酸盐正电子发射断层扫描/计算机断层扫描对嵌合抗原受体T细胞进行动态成像

T cells genetically engineered to express chimeric antigen receptors (CAR) have shown unprecedented results in pivotal clinical trials for patients with B ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

肝细胞癌原位小鼠模型中的无创PET/MR成像

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

A Dual Tracer PET-MRI Protocol for the Quantitative Measure of Regional Brain Energy Substrates Uptake in the Rat

We present a method for comparing the uptake of the brain&#39;s two key energy substrates: glucose and ketones (acetoacetate [AcAc] in this case) in the rat. The developed method is a small-animal positron emission tomography (PET) protocol, in which&#160;11C-AcAc and 18F-fluorodeoxyglucose (18F-FDG) are injected sequentially in each animal. This dual tracer PET acquisition is possible because of the short half-life of 11C (20.4 min). The rats also undergo a magnetic resonance imaging (MRI) acquisition seven days before the PET protocol. Prior to image analysis, PET and MRI images are coregistered to allow the measurement of regional cerebral uptake (cortex, hippocampus, striatum, and cerebellum). A quantitative measure of 11C-AcAc and 18F-FDG brain uptake (cerebral metabolic rate; &#956;mol/100 g/min) is determined by kinetic modeling using the image-derived input function (IDIF) method. Our new dual tracer PET protocol is robust and flexible; the two tracers used can be replaced by different radiotracers to evaluate other processes in the brain. Moreover, our protocol is applicable to the study of brain fuel supply in multiple conditions such as normal aging and neurodegenerative pathologies such as Alzheimer&#39;s and Parkinson&#39;s diseases.

Small-animal positron emission tomography enables the assessment of the brain&#39;s two main energy substrates:&#160;glucose and ketones. In the present method, 11C-acetoacetate and 18F-fluorodeoxyglucose are injected sequentially in each animal, and their uptake is measured quantitatively in specific brain regions determined from the magnetic resonance images.

We present a method for comparing the uptake of the brain&#39;s two key energy substrates: glucose and ketones (acetoacetate [AcAc] in this case) in the ...

神经科学

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to return to normal in a large majority of patients2, which is mainly due to current lack of clinical treatment for acute stroke. This necessitates understanding the physiological effects of cerebral ischemia on brain tissue over time and is a major area of active research. Towards this end, experimental progress has been made using rats as a preclinical model for stroke, particularly, using non-invasive methods such as 18F-fluorodeoxyglucose (FDG) coupled with Positron Emission Tomography (PET) imaging3,10,17. Here we present a strategy for inducing cerebral ischemia in rats by middle cerebral artery occlusion (MCAO) that mimics focal cerebral ischemia in humans, and imaging its effects over 24 hr using FDG-PET coupled with X-ray computed tomography (CT) with an Albira PET-CT instrument. A VOI template atlas was subsequently fused to the cerebral rat data to enable a unbiased analysis of the brain and its sub-regions4. In addition, a method for 3D visualization of the FDG-PET-CT time course is presented. In summary, we present a detailed protocol for initiating, quantifying, and visualizing an induced ischemic stroke event in a living Sprague-Dawley rat in three dimensions using FDG-PET.

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Brain damage resulting from cerebral ischemia may be non-invasively imaged and studied in rats using pre-clinical positron emission tomography coupled with the injectable radioactive probe, 18F-fluorodeoxyglucose. Further, the use of modern software tools that include volume of interest (VOI) brain templates dramatically increase the quantitative information gleaned from these studies.

无创成像和脑缺血分析生活中的大鼠使用正电子发射断层扫描<sup&gt; 18</sup&gt; F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

医学

Preparing a 68Ga-labeled Arginine Glycine Aspartate (RGD)-peptide for Angiogenesis

The &#945;v&#946;3 integrin is a heterodimeric adhesion molecule involved in tumor cell migration and angiogenesis. The integrin is overexpressed in angiogenic tumor endothelial cells, where it typically has a low concentration. This specific expression of &#945;v&#946;3 makes it a valid biomarker for antiangiogenic and imaging drugs. As a functional imaging modality, positron emission tomography (PET) provides information about biochemical and physiological changes in vivo, due to its unique high sensitivity at the nanomolar scale. Hence, radiometal-based PET radiopharmaceuticals have received great attention for the non-invasive quantification of tumor angiogenesis. This paper provides a systemic protocol to prepare a new radiometal-labeled peptide for the evaluation of angiogenesis. This protocol contains information about radiochemical reliability, lipophilicity, cell uptake, serum stability, and pharmacokinetic properties. The 68Ga-RGD-peptide is one of the representative PET ligands toward &#945;v&#946;3 integrin. Here, we introduce a protocol to prepare a 68Ga-RGD-peptide and the evaluation of its biological efficacy.

Preparing a 68Ga-labeled Arginine Glycine Aspartate RGD-peptide for Angiogenesis

The &#945;v&#946;3 integrin is a type of adhesion protein that is highly expressed on activated endothelial cells undergoing angiogenesis. Thus, evaluating the integrity of the integrin is of great interest in oncology. Here, we introduce a method to prepare 68Ga-labeled radiopeptides and a method to assess its biological effectiveness.

68加标记精氨酸甘氨酸 (rgd)-肽的血管生成

The &#945;v&#946;3 integrin is a heterodimeric adhesion molecule involved in tumor cell migration and angiogenesis. The integrin is overexpressed in ...

A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection

Renal allograft rejection limits the long-term survival of patients after renal transplantation. Rat orthotopic renal transplantation is an essential model to investigate the mechanism of renal allograft rejection in pre-clinical studies and could aid in the development of novel approaches to improve the long-term survival of renal allografts. Donor kidney implantation in rat orthotopic renal transplantation is commonly performed by end-to-side anastomosis to recipients' aorta and inferior vena cava. In this model, the donor's kidney was implanted using end-to-end anastomosis to the recipients' renal artery and renal vein. The donor's ureter was anastomosed to the recipient's bladder in an end-to-side 'tunnel' method. This model contributes to better healing of ureter-bladder anastomosis and increases the recipients' survival by avoiding interference with blood supply and venous reflux of the lower body. This model can be used to investigate the mechanisms of acute and chronic immune and pathologic rejection of renal allografts. Here, the study describes the detailed protocols of this orthotopic renal transplantation between rats.

The rat orthotopic renal transplantation model contributes to investigating the mechanism of renal allograft rejection. The current model increases the recipients' survival without interference with blood supply and venous reflux of the lower body using an end-to-end anastomosis of kidney implantation and an end-to-side "tunnel" method of ureter-bladder anastomosis.

一种用于肾同种异体排斥的大鼠原位肾移植模型

Renal allograft rejection limits the long-term survival of patients after renal transplantation. Rat orthotopic renal transplantation is an essential ...

High-Resolution Cardiac Positron Emission Tomography/Computed Tomography for Small Animals

Positron emission tomography (PET) and computed tomography (CT) are among the most employed diagnostic imaging techniques, and both serve in understanding cardiac function and metabolism. In preclinical research, dedicated scanners with high sensitivity and high spatio-temporal resolution are employed, designed to cope with the demanding technological requirements posed by the small heart size and very high heart rates of mice and rats. In this paper, a bimodal cardiac PET/CT imaging protocol for experimental mouse and/or rat models of cardiac diseases is described, from animal preparation and image acquisition and reconstruction to image processing and visualization.
In particular, the 18F-labeled fluorodeoxyglucose ([18F]FDG)-PET scan allows for the measurement and visualization of glucose metabolism in the different segments of the left ventricle (LV). Polar maps are convenient tools to display this information. The CT part consists of a time-resolved 3D reconstruction of the entire heart (4D-CT) using retrospective gating without electrocardiography (ECG) leads, allowing the morphofunctional evaluation of the LV and the subsequent quantification of the most important cardiac function parameters, such as ejection fraction (EF) and stroke volume (SV). Using an integrated PET/CT scanner, this protocol can be executed within the same anesthesia induction without the need to reposition the animal between different scanners. Hence, PET/CT can be seen as a comprehensive tool for the morphofunctional and metabolic evaluation of the heart in several small animal models of cardiac diseases.

Here, we present an experimental imaging protocol for the quantification of cardiac function and morphology using high-resolution positron emission tomography/computed tomography for small animals. Both mice and rats are considered, discussing the different requirements of computed tomography contrast agents for the two species.

小动物高分辨率心脏正电子发射断层扫描/计算机断层扫描

Positron emission tomography (PET) and computed tomography (CT) are among the most employed diagnostic imaging techniques, and both serve in understanding ...

FDG Based PET/CT Scan: A Method of Using Radiotracer for Non-invasive Imaging of Tumor Metabolism

Source: Momcilovic M. et al., <a href="https://www.jove.com/v/57167/utilizing-18f-fdg-petct-imaging-quantitative-histology-to-measure">Utilizing&#160;18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer.</a> J. Vis. Exp. (2018)
This article describes multimodality imaging of cancerous tissues with PET and CT diagnostic imaging by using a radiotracer 18F-FDG. Cancerous tissue with high glycolytic activity uptakes more 18F-FDG.

基于 FDG 的 PET/CT 扫描:一种使用放射性示踪剂进行肿瘤代谢无创成像的方法

Source: Momcilovic M. et al., Utilizing&#160;18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in ...

大鼠肾移植后急性排斥反应的非侵入性的成像使用

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