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，AR AR使用FDG-PET 3。由于FDG-PET成像需要相对少量的活性和成像要么移植患者和/或肾功能受损的患者不伴有严重并发症的风险增加，这种方法可以很容易地转让融入日常临床常规。 然而，一个人要记住，FDG是相当非特异性示踪剂评估区域代谢活动。因此，移植可能导致感染或肿瘤以及假阳性结果。在肾实质评估FDG摄取FDG进入肾盂引流可能是一个问题。因此，我们选择了后期采集时间3小时后注射，以减少在肾脏引起的肾脏排泄FDG示踪剂积累。此外，肾盂量化肾FDG摄取时，必须谨慎地排除。根据本协议PET可用于非侵入性检测AR，以区别于ATN和CSA，并执行串行调查为后续处理效率3，4，6或评价。 PET使用18 F-氟化物，也是有用的，以评估（分体）肾功能由计算的氟化肾间隙公布BEF矿石5。 同种异体肾移植模型使用LBN F1捐助者和受体鼠刘易斯是调查急性细胞移植排斥反应的理想模型。在没有免疫抑制肾脏同种异体移植物的发展。根据至BANFF的分类7的AR的典型的组织学标志。此外，根据所选择的方式（单向与binephrectomized的移植3 8-10）代谢数据可以评价为监测移植肾功能。由于免疫抑制治疗的情况下，伤口或全身性感染的老鼠是极其罕见的。这种模式常见的手术并发症包括血管狭窄，大多出现在进入主动脉或ICV收件人移植血管的插入点。这可能会导致缺血或接枝血栓形成。人们可以使用更长的切口主动脉和下腔静脉，避免这种并发症。有时发现尿毒症，由于移植失败或ureter泄漏造成输尿管膀胱输尿管坏死或断开。如果出现尿毒症， 如淡漠，丧失食欲或体重增加自发的迹象，应立即处死动物。

<table border="1">
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 <td>Equipment</td>
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 <td>Mathieu Needle Holder - 14 cm</td>
 <td>Fine Science Tools</td>
 <td>12010-14</td>
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 <td>Castroviejo Micro Needle Holder - 9 cm</td>
 <td>Fine Science Tools</td>
 <td>12061-01</td>
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 <td>Surgical Scissors - Sharp_Blunt</td>
 <td>Fine Science Tools</td>
 <td>14001-12</td>
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 </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大量的葡萄糖代谢活跃的细胞。但是，如果是后者取代的葡萄糖，这将堆积在细胞内，可以测量和量化PET。 PET图像代表的PET图像的动态全身人一系列的收购logeneically移植大鼠尾静脉注射后30活度FDG（最大后验地图投影，180分钟PI）（ 图2）。发现一个典型的FDG分布在大脑，心脏，骨髓和哈氏腺具有独特的生理积累。此外，免费滤FDG积聚在尿路。在接受肾移植AR（黄色圆圈，左肾实质）高度积累FDG一个最大POD4，而原生肾（绿色圆圈，右肾）不显示任何积累。由于肾盂可以包含免费淘汰FDG，它被排除在进一步的测量。图3。 定量评价对于定量评价图像重建，感兴趣体积肾脏周围的手动追溯到18 F-氟灌注FDG图像投射。排除后肾聚乙烯的LVIS意味着在肾实质FDG摄取量（％ID±SEM）总数量的比例计算。肾开发AR显着增加FDG积聚POD4（0.8±0.06％）相比，本地对照组（0.2±0.02％），或syngeneically移植肾脏（0.37±0.04％）。此外，阿肯色州两个主要的鉴别诊断，即急性肾小管坏死缺血/再灌注损伤（IRI）（0.31±0.02％）和急性毒性calcineurin抑制剂（CSA）（0.16±0.01％）一样，没有表现出升高FDG积聚因此，可以区分从AR（取自3的 图3）。 <img src="/files/ftp_upload/4240/4240fig1.jpg" alt="图1"/> 图1。组织学检查。迹象急性排斥反应，即肾小球肾炎，小管炎，内皮炎，和移植物浸润，被发现在移植组（ATX）和完全控制肾脏（CTR）缺席。 <img src="/files/ftp_upload/4240/4240fig2.jpg" alt="图2"/> 图2。 FDG-PET图像，代表PET图像的动态全身的一系列收购一个allogeneically大鼠移植。控制肾脏（绿色圆圈）相比，肾移植（黄色圆圈）FDG最大POD4积累的实质。由于肾盂可以包含自由淘汰FDG排除测量。图3。 <img src="/files/ftp_upload/4240/4240fig3.jpg" alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/4240/4240fig3highres.jpg"/> 图3。定量评价。急性排斥反应的检测通过测量葡萄糖的％ID。肾移植（ATX）表现出显着较高的FDG积累比对照组肾脏（CTR），syngeneically同种异体移植物（STX），肾脏急性肾小管坏死（ATN）或肾脏急性calcineurin抑制剂毒性（CSA）与最大POD4（ATX:0.8±0.06％的点击率:0.2±0.02 ％，STX:0.37±0.04％，ATN:0.31±0.02％，CSA:0.16±0.01％）。图3。

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

Nanyang Technological University

Research

Education

JoVE Journal

JoVE Core

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

Non-invasive Assessment of Microvascular and Endothelial Function

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of cardiovascular disease. Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. Percent capillary recruitment is assessed by dividing the increase in capillary density induced by postocclusive reactive hyperemia (postocclusive reactive hyperemia capillary density minus baseline capillary density), by the maximal capillary density (observed during passive venous occlusion). Percent perfused capillaries represents the proportion of all capillaries present that are perfused (functionally active), and is calculated by dividing postocclusive reactive hyperemia capillary density by the maximal capillary density. Both percent capillary recruitment and percent perfused capillaries reflect the number of functional capillaries. The forearm blood flow (FBF) technique provides accepted non-invasive measures of endothelial function: The ratio FBFmax/FBFbase is computed as an estimate of vasodilation, by dividing the mean of the four FBFmax values by the mean of the four FBFbase values. Forearm vascular resistance at maximal vasodilation (FVRmax) is calculated as the mean arterial pressure (MAP) divided by FBFmax. Both the capillaroscopy and forearm techniques are readily acceptable to patients and can be learned quickly.
The microvascular and endothelial function measures obtained using the methodologies described in this paper may have future utility in clinical patient cardiovascular risk-reduction strategies. As we have published reports demonstrating that microvascular and endothelial dysfunction are found in initial stages of hypertension including prehypertension, microvascular and endothelial function measures may eventually aid in early identification, risk-stratification and prevention of end-stage vascular pathology, with its potentially fatal consequences.

Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. The forearm blood flow technique provides accepted non-invasive measures of endothelial function.

非侵入性的评估微血管和内皮功能

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of ...

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 ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

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 ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

新的治疗肠道病变的疗效，使用活体小鼠的串行内镜成像的非侵入性评估

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

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 ...

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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