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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+and+chronic+nephropathy:+a+systematic+review+of+the+literature.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplantation+in+the+rat.">Kidney transplantation in the rat.</a> Nature. 194, 309-310 (1962).</li><li>Reuter, S., Schn&#246;ckel, U., Schr&#246;ter, R., Schober, O., Pavenst&#228;dt, H., Sch&#228;fers, M., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+imaging+of+acute+renal+allograft+rejection+in+rats+using+small+animal+F-FDG-PET.">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., Schn&#246;ckel, U., Edemir, B., Schr&#246;ter, R., Kentrup, D., Pavenst&#228;dt, H., Schober, O., Schlatter, E., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+of+noninvasive+serial+assessment+of+acute+renal+allograft+rejection+by+18F-FDG+PET+to+monitor+treatment+efficiency.">Potential of noninvasive serial assessment of acute renal allograft rejection by 18F-FDG PET to monitor treatment efficiency.</a> J. Nucl. Med. 51, 1644-1652 (2010).</li><li>Schn&#246;ckel, U., Reuter, S., Stegger, L., Schlatter, E., Sch&#228;fers, K. P., Hermann, S., Schober, O., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+18F-fluoride+small+animal+PET+to+noninvasively+assess+renal+function+in+rats.">Dynamic 18F-fluoride small animal PET to noninvasively assess renal function in rats.</a> Eur. J. Nucl. Med. Mol. Imaging. 35, 2267-2274 (2008).</li><li>Grabner, A., Schn&#246;ckel, U., Kentrup, D., Sch&#228;fers, M., Reuter, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+for+Non-Invasive+Molecular+Imaging+of+Acute+Allograft+Rejection+by+Gamma+Scintigraphy+and+Positron+Emission+Tomography.">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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Banff+97+working+classification+of+renal+allograft+pathology.">The Banff 97 working classification of renal allograft pathology.</a> Kidney Int. 55, 713-723 (1999).</li><li>Edemir, B., Reuter, S., Borgulya, R., Schr&#246;ter, R., Neugebauer, U., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+modulates+gene+expression+in+the+collecting+duct.">Acute rejection modulates gene expression in the collecting duct.</a> J. Am. Soc. Nephrol. 19, 538-546 (2008).</li><li>Velic, A., Gabri&#235;ls, G., Hirsch, J. R., Schr&#246;ter, R., Edemir, B., Paasche, S., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+after+rat+renal+transplantation+leads+to+downregulation+of+Na++and+water+channels+in+the+collecting+duct.">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><li>Reuter, S., Velic, A., Edemir, B., Schr&#246;ter, R., Pavenst&#228;dt, H., Gabri&#235;ls, G., Bleich, M., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+role+of+NHE-3+inhibition+in+rat+renal+transplantation+undergoing+acute+rejection.">Protective role of NHE-3 inhibition in rat renal transplantation undergoing acute rejection.</a> Pflugers Arch. 456, 1075-1084 (2008).</li></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 border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</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></td>
 <td></td>
 <td></td>
 <td>Reagent</td>
 </tr>
 <tr>
 <td>Isoflurane (e.g. Forene 100% v/v)</td>
 <td>Abott</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>cutane antiseptic (e.g. Octeniderm) </td>
 <td>Sch&uuml;lke</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Povidone Iodine (e.g. Betaisodona)</td>
 <td>Mundipharma</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>ophthalmic ointment (e.g. Bepanthen)</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprenorphin (e.g. Temgesic)</td>
 <td>RB Pharmaceuticals</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>HTK perfusion solution (e.g. CUSTODIOL HTK)</td>
 <td>Dr. Franz K&ouml;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&ouml;mel</td>
 <td>14739</td>
 <td></td>
 </tr>
 <tr>
 <td>Canula (e.g. Microlance 3, 27G &frac34;)</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

Research

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JoVE Journal

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Biology

Medicine

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

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

婴幼儿的脑代谢和血流动力学的非侵入式光学测量

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

科研

医学

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Intravital imaging has emerged as an indispensable tool in biological research. In the process, many imaging techniques have been developed to study different biological processes in animals non-invasively. However, a major technical limitation in existing intravital imaging modalities is the inability to combine non-invasive, longitudinal imaging with single-cell resolution capabilities. We show here how transplantation into the anterior chamber of the eye circumvents such significant limitation offering a versatile experimental platform that enables non-invasive, longitudinal imaging with cellular resolution in vivo. We demonstrate the transplantation procedure in the mouse and provide representative results using a model with clinical relevance, namely pancreatic islet transplantation. In addition to enabling direct visualization in a variety of tissues transplanted into the anterior chamber of the eye, this approach provides a platform to screen drugs by performing long-term follow up and monitoring in target tissues. Because of its versatility, tissue/cell transplantation into the anterior chamber of the eye not only benefits transplantation therapies, it extends to other in vivo applications to study physiological and pathophysiological processes such as signal transduction and cancer or autoimmune disease development.

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

A new approach combining intraocular transplantation and confocal microscopy enables longitudinal, non-invasive real-time imaging with single-cell resolution within grafted tissues in vivo. We demonstrate how to transplant pancreatic islets into the anterior chamber of the mouse eye.

纵向，非侵入性的移植到眼睛的前房<em在体内</em&gt;单细胞分辨率的实时成像

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

MRI and PET in Mouse Models of Myocardial Infarction

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ideal model for testing novel therapeutic strategies.
In vivo magnetic resonance imaging (MRI) gives excellent views of the heart noninvasively with clear anatomical detail, which can be used for accurate functional assessment. Contrast agents can provide basic measures of tissue viability but these are nonspecific. Positron emission tomography (PET) is a complementary technique that is highly specific for molecular imaging, but lacks the anatomical detail of MRI. Used together, these techniques offer a sensitive, specific and quantitative tool for the assessment of the heart in disease and recovery following treatment.
In this paper we explain how these methods are carried out in mouse models of acute myocardial infarction. The procedures described here were designed for the assessment of putative protective drug treatments. We used MRI to measure systolic function and infarct size with late gadolinium enhancement, and PET with fluorodeoxyglucose (FDG) to assess metabolic function in the infarcted region. The paper focuses on practical aspects such as slice planning, accurate gating, drug delivery, segmentation of images, and multimodal coregistration. The methods presented here achieve good repeatability and accuracy maintaining a high throughput.

We describe how to perform MRI and PET imaging of the mouse heart. The protocol is tailored to assess treatment efficacy in models of myocardial infarction and heart failure.

MRI和PET心肌梗死小鼠模型

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ...

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 ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

小鼠模型的非侵入性成像检测和监测卵巢癌复发

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

鼠肾移植:移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

确定葡萄糖代谢动力学研究<sup&gt; 18</sup&gt; F-FDG微PET / CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

脂肪组织干细胞移植对大鼠部分结肠切除术后渗漏的减少

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of disorders of consciousness, disrupted higher brain function, and verbal disturbances. As a consequence of an insufficient ability to communicate, objective evaluations are needed from family members, medical staff, and caregivers. One such evaluation is the assessment of functioning brain areas. Recently, multimodal brain imaging has been used to explore the function of damaged brain areas. [18F]-fluorodeoxyglucose positron emission tomography-computed tomography ([18F]FDG-PET/CT) is a successful tool for examining brain function. However, the assessment of brain glucose metabolism based on [18F]FDG-PET/CT is not standardized and depends on several varying parameters, as well as the patient&#39;s condition. Here, we describe a series of semiquantitative assessment protocols for a region-of-interest (ROI) image analysis using self-produced [18F]FDG tracers in patients with sTBI. The protocol focuses on screening the participants, preparing the [18F]FDG tracer in the hot lab, scheduling the acquisition of [18F]FDG-PET/CT brain images, and measuring glucose metabolism using the ROI analysis from a targeted brain area.

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

[18F]-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography is useful for studying glucose metabolism related to brain function. Here, we present a protocol for an [18F]FDG tracer set-up and semiquantitative assessment of the region-of-interest analysis for targeted brain areas associated with clinical manifestations in patients with severe traumatic brain injury.

[18f] fdg 示踪剂在重型颅脑损伤患者中的半定量评价

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...