Name: non-invasive imaging of acute allograft rejection after rat renal transplantation using 18f-fdg pet
Uploaded: 2013-04-28T14:00:00.000+00:00
Duration: 17 min 13 s
Description: Watch this Scientific Journal Video about Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET at JoVE.com

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>

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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 ...

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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

Machine Learning Algorithms for Early Detection of Bone Metastases in an Experimental Rat Model

Machine learning (ML) algorithms permit the integration of different features into a model to perform classification or regression tasks with an accuracy exceeding its constituents. This protocol describes the development of an ML algorithm to predict the growth of breast cancer bone macrometastases in a rat model before any abnormalities are observable with standard imaging methods. Such an algorithm can facilitate the detection of early metastatic disease (i.e., micrometastasis) that is regularly missed during staging examinations.
The applied metastasis model is site-specific, meaning that the rats develop metastases exclusively in their right hind leg. The model&#8217;s tumor-take rate is 60%&#8211;80%, with macrometastases becoming visible in magnetic resonance imaging (MRI) and positron emission tomography/computed tomography (PET/CT) in a subset of animals 30 days after induction, whereas a second subset of animals exhibit no tumor growth.
Starting from image examinations acquired at an earlier time point, this protocol describes the extraction of features that indicate tissue vascularization detected by MRI, glucose metabolism by PET/CT, and the subsequent determination of the most relevant features for the prediction of macrometastatic disease. These features are then fed into a model-averaged neural network (avNNet) to classify the animals into one of two groups: one that will develop metastases and the other that will not develop any tumors. The protocol also describes the calculation of standard diagnostic parameters, such as overall accuracy, sensitivity, specificity, negative/positive predictive values, likelihood ratios, and the development of a receiver operating characteristic. An advantage of the proposed protocol is its flexibility, as it can be easily adapted to train a plethora of different ML algorithms with adjustable combinations of an unlimited number of features. Moreover, it can be used to analyze different problems in oncology, infection, and inflammation.

This protocol was designed to train a machine learning algorithm to use a combination of imaging parameters derived from magnetic resonance imaging (MRI) and positron emission tomography/computed tomography (PET/CT) in a rat model of breast cancer bone metastases to detect early metastatic disease and predict subsequent progression to macrometastases.

实验鼠模型中骨代谢的早期检测机器学习算法

Machine learning (ML) algorithms permit the integration of different features into a model to perform classification or regression tasks with an accuracy ...

科研

癌症研究

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 ...

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

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 ...

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 ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

大鼠原位肾移植: 一种新的简化的手术方法

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

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 ...

Liver Transplantation in Rats and Analyzing Allograft Patency

Heterotopic Auxiliary Whole Liver Rat Transplant Model Utilizing a Hepaticoureterostomy for Allograft Rejection Studies

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat liver transplantation (LTx) protocols typically use an orthotopic model where the recipients&#39; native liver is removed and replaced with a donor liver. This technically demanding surgical procedure requires advanced micro-surgical skills and is further complicated by lengthy anhepatic and lower body ischemia times. This prompted the development of a less complicated heterotopic method that can be performed faster with no anhepatic or lower body ischemia time, reducing post-surgery stress for the recipient animal.
This heterotopic LTx protocol includes two main steps: excising the liver from the donor rat and transplanting the whole liver into the recipient rat. During the excision of the donor liver, the surgeon ligates the supra-hepatic vena cava (SHVC) and hepatic artery (HA). On the recipient side, the surgeon removes the left kidney and positions the donor liver with the portal vein (PV), infra-hepatic vena cava (IHVC), and bile duct facing the renal vessels. Further, the surgeon anastomoses the recipient&#39;s renal vein end to end with the IHVC of the liver and arterializes the PV with the renal artery using a stent. A hepaticoureterostomy is utilized for biliary drainage by anastomosing the bile duct to the recipient&#39;s ureter, permitting the discharge of bile via the bladder.
The average duration of the transplantation was 130 min, cold ischemia duration was around 35 min, and warm ischemia duration was less than 25 min. Hematoxylin and eosin histology of the auxiliary liver from syngeneic transplants showed normal hepatocyte structure with no significant parenchymal alterations 30 days post-transplant. In contrast, 8-day post-transplant allogeneic graft specimens demonstrated extensive lymphocytic infiltration with a Banff Schema rejection activity index score of 9. Therefore, this LTx method facilitates a low morbidity rejection model alternative to orthotopic LTx.

The rat heterotopic auxiliary liver transplant protocol described here offers a practical investigational tool for exploring mechanisms of hepatic allograft rejection. This model helps to alleviate the surgical hurdles and animal stress of orthotopic liver transplantation in rats.

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat ...

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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