作者感谢捐助者的家庭和组织的专家协助参与这项研究和玛丽亚·马蒂诺和伊琳娜Kusmartseva器官采购。这项工作是由青少年糖尿病研究基金会（MC-T.，马萨诸塞州）在对糖尿病患者的胰腺器官捐赠者的网络支持。

1。设置<ol><li>石蜡块，用铅笔或自动录像带打印机手动标签的录像带。包括捐赠标识符（CaseID），样本类型，等分数目，以及任何额外的唯一标识（案例表， <a href="http://jove.com/files/ftp_upload/4039/4039appendix1.doc" target="_blank">附录1</a> ）。 </li><li>标签华侨城模具为永久性标记录音带。剪下铝箔5-10厘米的长方形。 </li><li>打印或手工标签永久性标记所有小瓶和管。包括足够瓶碎单元冷冻样品和新鲜出货的RPMI媒体管。为冷冻小瓶与RNAlater单元的，加1毫升RNAlater每个离心管。 </li><li>在生物安全罩和柜台设置清扫与消毒清扫工具（镊子，手术刀，刀片，纱布垫（4×4））以及磁带和华侨城模具板在使用顺序排列。 </li><li>通过把准备冻结浴干冰在一个中型泡沫塑料箱（10×10厘米）至少有3厘米的深度，并添加异戊烷，直到完全覆盖干冰。 </li><li>可选:填写一个小杜瓦液氮容器。 </li><li>如果需要电子显微镜（EM）胰腺，准备加入1.25毫升16％多聚甲醛，8毫升1.25％戊二醛，7.5毫升0.1M PBS TAAB固定液。准备加入2.5毫升16％多聚甲醛7.5毫升0.1M PBS Lowicryl固定液。 </li><li>准备用于分离细胞的解决方案。以500毫升瓶装的DMEF（谷氨酰胺），遵循无菌技术，同时添加以下: <ol><li>胎牛血清（FBS），附加50毫升，最后的10％。 </li><li>青霉素/链霉素（PEN /链球菌，1万U /毫升/ 10000微克/毫升股票） - 添加5毫升笔/链球菌股票的的RPMI终浓度为100 U / ml的/ 100ug/ml的。还可以添加5毫升至500毫升瓶举行淋巴结或脾样本使用时DPBS。 </li></ol></li></ol> 2。人员<oL&gt; <li>三名工作人员，但该程序可以有一个训练有素的个人进行过程优化完成。两个人一般可以在90分钟内完成的过程。实际工期是依赖于接受组织的数量和减轻胰腺淋巴结的发现。细胞隔离的样品在4日举行°C，直到完成组织剥离，而并行处理的血清标本。 </li><li>病理学家，病理学助理，或相当于一个经验丰富的工作人员，导致了组织处理和执行主要包括胰头部（S）和收获胰腺淋巴结清扫。最初的努力是针对胰腺检查胰周淋巴结的脂肪。 </li><li>实验室技师或相当于工作人员培训，组织修剪，其主要职责是衡量固定及冷冻块和单元冷冻至五胰腺和地区，修剪组织IALS。接收从病理学助理十二指肠和脾脏和处理，而这些器官的病理学助理处理胰腺。 </li><li>实验室助理工作人员，帮助别人，包括块，小瓶标签，样品切碎的冷冻管，血清处理。 </li></ol> 3。胰腺解剖<ol><li>取出十二指肠和脾脏，用钝，锐性剥离，清除多余的脂肪，血管和神经，胰腺</li><li>将清洁用纸巾覆盖在解剖板胰腺。胰腺的前表面上沾了蓝墨水和蔓延的棉签。吸干多余的。 </li><li>可选:沾在黑色墨水和传播新的胰腺后表面撒施。与前表面面临解剖员返回胰腺正常的方向。 </li><li>切成3个地区的胰腺头部，身体，尾巴（ 图1）。削减HEA之间的交界处d和身体，然后分为身体和尾巴〜等份的剩余部分。 </li><li>权衡各地区胰腺和记录重量（案例相关检验表（ <a href="http://jove.com/files/ftp_upload/4039/4039appendix1.doc" target="_blank">附录1</a> ））。 </li><li>胰腺中的一横的"面包"的方式（ 图1）各地区组（约0.5厘米）。 </li><li>使用碎样本地区之间的两个路口的部分。根据胰腺大小，从体尾交界地区的样本可能很难获得石蜡块的数量，在这种情况下，可以降低获得碎样品或从胰头体地区唯一的样品，可以得到。标签冷冻管头部或身体来表示近端地区的交界处。 <ol><li>百果片和冷冻管（≥1克每瓶）之间均匀地划分。迅速冻结在液氮或干冰RNALater小瓶 - 异戊烷浆，​​然后转移ţOA -80°C冰箱。 </li><li>保持与RNALater小瓶30分钟，在室温下，以便平衡，混音，并迅速冻结在液氮或干冰戊烷浆，然后转移到-80℃冰箱。 </li></ol></li><li>删除石蜡和OCT块石蜡（ 图1）开始交替部分。 </li><li>瘦到正确的横截面。 </li><li> 〜1.5×1.5×0.5厘米的录音带的部分地方。如果横截面过大，减少了一半（ 图2b）。蓝色标签为"A"和后半段为"B"的前半录音带。 </li><li>如果件仍然过大，每节垂直削减前切（ 图2c）。公元标签录音带以顺时针的方式。修剪进一步需要，以适应在录音带的片断。 </li><li>数块的顺序，在区域范围内，最内侧的部分开始。 </li><li>对于石蜡块，位置C所以标签assettes是左边和地方组织成磁带，保持其方向。关闭录像带盖和转移到一个容器〜500毫升10％的中性福尔马林（NBF）。开始计时当最后录像带被放置在固定液固定。 </li><li>十月块，倒入一层薄薄的华侨城媒体prelabeled模具1模具一定要保持方向，并覆盖组织与华侨城奠定了组织。迅速沉浸在干冰华侨城模具 - 戊烷浆〜1分钟。从泥浆和删除临时存储干冰或-80°C下长期储存的冷冻。 </li><li>获得头体交界处，从电子显微镜（EM）〜0.5×0.5厘米的薄片。片在TAAB固定液和Lowicryl固定液的其他片的地方之一。储存在4°C两种类型的固定液与样品至少48小时和/或无限期直至EM进一步处理。 </li></ol> 4。脾，淋巴结节点Dissections和十二指肠黏膜夹层<ol><li>从脂肪中分离胰腺癌淋巴结（兹罗提）和结缔组织修剪所有的节点舱。在无菌细胞培养DMEF菜的地方。根据分离的总数，分为新鲜的出货量，冻存细胞隔离，石蜡和/或十月块，小瓶分装兹罗提。剁碎切成不超过0.5厘米的大节点。 </li><li>剁碎成小立方体（约0.5厘米）的非胰腺淋巴结和脾脏。新鲜样品供货或冻存细胞隔离，根据起始原料的金额固定，华侨城媒体，或培养液中的地方。 </li><li>打开十二指肠，轻轻擦拭干净，用湿纱布粘膜，十二指肠粘膜。削减部分的黏膜石蜡和/或冷冻十月块和碎小瓶样品，与RNAlater。冻结为其他样品。 </li></ol> 5。 WRAP十月预标记的铝箔和地方十月块和块<html>在长期储存箱小瓶在-80°C。 6。提交固定标本，石蜡处理<ol><li>使用自动的石蜡处理器或使用手动计时，修复组织了16个小时（范围为20±4小时）。工艺蜡块，用自动处理器（ <a href="http://jove.com/files/ftp_upload/4039/4039appendix2.doc" target="_blank">附录2</a> ）。 </li><li>录像带标签左侧的解剖员放置在准确定位的嵌入部分。嵌入与十二指肠黏膜切面允许检查黏膜垂直粘膜下层。 </li></ol> 7。清洁夹层领域和消毒。洗清扫工具，并重新消毒。生物医学废物贮存，放置在福尔马林固定液残留的人体组织。根据当地法规丢弃在一个月内的生物医学废物。 8。更新的样品库存系统ve_title"&gt; 9代表结果。 此过程将处理整个器官，包括三大区域，即，头部，身体和尾部2小时内划分的抽样代表一个完整的人类胰腺。钩突的地区，发现在后头部区域，包括在头部解剖。在没有糖尿病的器官捐赠者的平均胰腺重量为82.4克（52.7 - 139.0克）。胰腺区域重分别为约等于（ 图3）。 这个过程也可以让重建整个胰腺利用连续石蜡和华侨城块的彩色幻灯片。多种格式允许的最大利用率，为当前和未来的技术是可行的。当前nPOD情况表（ <a href="http://jove.com/files/ftp_upload/4039/4039appendix1.doc" target="_blank">附录1</a> ），用于记录回收样品和随后的PROCessing由等分的种类和数量。 <img alt="图1" src="/files/ftp_upload/4039/4039fig1.jpg" /> 图1。程序的总体方案。整个胰腺进行处理，同时保持解剖方向。头地区是在优越的上下轴，由于腹胰叶后的地区，其中包括钩突地区的存在。孵化面积在路口表示用于碎样本在冷冻管的地区。磷，石蜡，O，华侨城。 <img alt="图2" src="/files/ftp_upload/4039/4039fig2.jpg" /> 图2。刻字胰腺部分计划。横向胰腺切片可进一步分为两半或季度字母以顺时针的方式。 <img alt="图3" src="/files/ftp_upload/4039/4039fig3.jpg" /> 图3。胰腺重量从没有糖尿病的器官捐赠者。从成年捐赠者（&gt; 17岁）的完整胰腺称重，然后分为区域和区域的权重获得。数据是手段±标准差（n = 15）。

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没有利益冲突的声明。

此过程的目标是定义一个标准的胰腺处理，可以跨多个收集点统一，从而使不同群体得到的结果进行比较。剥离的方法是基于标准的注释主要胰腺区域内细分区域的额外步骤与手术病理做法。标准化的biospecimen处理方法是必需的，以便收集高品质的最佳做法biospecimen收集出版由国家组织，如国家癌症研究所6 biobanking体现样品。从捐助者代表糖尿病和适当的控制人口的各个阶段的人类胰腺样?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>清扫工具，包括22个刀片的手术刀</td><td>各个</td><td>各个</td><td></td></tr><tr><td>离心管（15和50毫升） </td><td> Fisher Scientific则</td><td> 05-539-5，430828 </td><td>不育</td></tr><tr><td> cryotubes（1.8毫升） </td><td>厂商VWR北美国</td><td> 89004-300 </td><td>无菌，条形码可选</td></tr><tr><td> RNAlater </td><td> Ambion公司</td><td> AM7021 </td><td></td></tr><tr><td>外科病理学油墨</td><td></td><td></td><td></td></tr><tr><td>录音带</td><td>奔驰医学</td><td>中科院ECO109 </td><td></td></tr><tr><td> 10％的中性福尔马林</td><td> Fisher Scientific则</td><td> 23-245-685 </td><td></td></tr><tr><td>华侨城冻结媒体</td><td> TissueTek </td><td> 4583 </td><td></td></tr><tr><td>模具</td><td> ThermoScientific </td><td> 58952 </td><td>各种规格</td></tr><tr><td>异戊烷</td><td> Fisher Scientific则</td><td> O3551-4 </td><td>华侨城低温浴</td></tr><tr><td>干冰</td><td></td><td></td><td>华侨城低温浴</td></tr><tr><td>液氮杜瓦瓶</td><td></td><td></td><td>抢购冷冻冷冻管</td></tr><tr><td>细胞培养皿（100毫米） </td><td>康宁</td><td> 430167 </td><td>不育</td></tr><tr><td>胎牛血清DMEF </td><td> ThermoScientific </td><td> SH30023.01 </td><td>谷氨酰胺包括无菌，储存冷藏， </td></tr><tr> &lt;TD&gt;胎牛血清</td><td> cellgro </td><td> 35-010-CV </td><td>无菌，冷冻分装店</td></tr><tr><td>青霉素/ /真菌</td><td> GIBCO公司</td><td> 15240-062 </td><td>无菌，冷冻分装店</td></tr><tr><td>键入9石蜡</td><td>理查德 - 阿兰科学</td><td> 8337 </td><td></td></tr><tr><td> 1XÐ-PBS </td><td> GIBCO公司</td><td> 14190-144 </td><td>不育</td></tr><tr><td> 16％多聚甲醛</td><td>新兴科学</td><td> 15710 </td><td>新鲜，商店冷藏</td></tr><tr><td> 8％的戊二醛</td><td>新兴科学</td><td> 16019 </td><td>新鲜，商店冷藏</td></tr><tr><td>生物安全等级2罩</td><td>各个</td><td></td><td></td></tr><tr><td>超低冰柜</td><td>各个</td><td></td><td></td></tr><tr><td>樱花纸盒打印机</td><td>樱花</td><td></td><td></td></tr><tr><td>樱花VIP300 </td><td>樱花</td><td></td><td>自动石蜡处理器的</td></tr><tr><td>石蜡包埋站</td><td> ThermoFisher </td><td></td><td></td></tr></tbody></table> 表1。特定的试剂和设备。

collection protocol for human pancreas

这解剖和取样程序开发为糖尿病（nPOD）的计划，以规范准备恢复从尸体器官捐献者的胰腺胰腺器官捐赠者的网络。胰腺分为3个主要地区整个横向轴内侧串行的横截面（头，身，尾）。交替部分用于固定石蜡和鲜冻块和残留样品剁碎单元冷冻样品准备，无论有或无RNase抑制剂，DNA，RNA或蛋白质隔离。胰腺的解剖过程的总体目标是保持解剖定位的同时，品尝整个胰腺。 在小岛组成，大小和数字方面的内分泌细胞的异质性是人类胰岛的报告相比，啮齿动物胰岛1。大多数人类胰岛胰头，身体和尾巴地区组成的胰岛素containinGβ-细胞的比例较低，其次是胰高血糖素的α细胞和生长抑素的δ-细胞。胰多肽含PP细胞生长素含小量细胞也存在，但在小数字。相比之下，钩突地区包含主要胰岛胰多肽含PP细胞组成。这些区域的胰岛变化，从发展的差异产生。胰腺发展的前肠和胃和十二指肠，腹侧叶动作和保险丝背3旋转后的腹侧和背侧胰芽。腹侧叶形成的头后部，包括钩突背叶而产生的其他器官。另据报道，尾部区域，具有较高的胰岛细胞密度比其他地区和背叶源性成分，在1型糖尿病的发生选择性萎缩区域胰腺癌的变化<s高达&gt; 4,5。 其他器官和组织往往是恢复从器官捐赠者和包括胰腺淋巴结，脾脏和非胰腺淋巴结。这些样品被收回冻存细胞的分离此外胰腺类似的格式。当近端十二指肠与胰腺，十二指肠粘膜可收集石蜡和冰冻块和碎单元冷冻的筹备工作。

Watch this Scientific Journal Video about Collection Protocol for Human Pancreas at JoVE.com

Collection Protocol for Human Pancreas

该视频演示了加工成多种存储格式的人类胰腺的解剖过程。解剖方向，保持整个胰腺地区允许区域胰岛成分和密度的定义。

人体胰腺的收集协议

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

Research

JoVE Journal

Medicine

14.7K 次观看.  University of Florida. 该视频演示了加工成多种存储格式的人类胰腺的解剖过程。解剖方向，保持整个胰腺地区允许区域胰岛成分和密度的定义。

视频: Collection Protocol for Human Pancreas

Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma 1,2. The dissected cells can be used for a variety of investigations, such as transcriptomic or proteomic studies, DNA assessment or chromosomal analysis 2,3. An especially challenging application of LMD is transcriptome analysis, which, due to the lability of RNA 4, can be particularly prominent when cells are dissected from tissues that are rich of RNases, such as the pancreas. A microdissection protocol that enables fast identification and collection of target cells is essential in this setting in order to shorten the tissue handling time and, consequently, to ensure RNA preservation. 
Here we describe a protocol for acquiring human pancreatic beta cells from surgical specimens to be used for transcriptomic studies 5. Small pieces of pancreas of about 0.5-1 cm3 were cut from the healthy appearing margins of resected pancreas specimens, embedded in Tissue-Tek O.C.T. Compound, immediately frozen in chilled 2-Methylbutane, and stored at -80 &deg;C until sectioning. Forty serial sections of 10 &mu;m thickness were cut on a cryostat under a -20 &deg;C setting, transferred individually to glass slides, dried inside the cryostat for 1-2 min, and stored at -80 &deg;C. 
Immediately before the laser microdissection procedure, sections were fixed in ice cold, freshly prepared 70% ethanol for 30 sec, washed by 5-6 dips in ice cold DEPC-treated water, and dehydrated by two one-minute incubations in ice cold 100% ethanol followed by xylene (which is used for tissue dehydration) for 4 min; tissue sections were then air-dried afterwards for 3-5 min. Importantly, all steps, except the incubation in xylene, were performed using ice-cold reagents - a modification over a previously described protocol 6. utilization of ice cold reagents resulted in a pronounced increase of the intrinsic autofluorescence of beta cells, and facilitated their recognition. For microdissection, four sections were dehydrated each time: two were placed into a foil-wrapped 50 ml tube, to protect the tissue from moisture and bleaching; the remaining two were immediately microdissected. This procedure was performed using a PALM MicroBeam instrument (Zeiss) employing the Auto Laser Pressure Catapulting (AutoLPC) mode. The completion of beta cell/islet dissection from four cryosections required no longer than 40-60 min. Cells were collected into one AdhesiveCap and lysed with 10 &mu;l lysis buffer. Each single RNA specimen for transcriptomic analysis was obtained by combining 10 cell microdissected samples, followed by RNA extraction using the Pico Pure RNA Isolation Kit (Arcturus). This protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their rapid and accurate recognition and collection. Further improvement of this procedure could enable the dissection of phenotypically different beta cells, with possible implications for better understanding the changes associated with type 2 diabetes.

Laser microdissection is a technique that allows the recovery of selected cells from minute amounts of parenchyma. Here we describe a protocol for acquiring human pancreatic islets from surgical specimens to be used for transcriptomic studies. Our protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their collection.

从手术标本的人类胰岛细胞激光显微切割的改进方法

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma 1,2. The dissected ...

科研

医学

Collection and Extraction of Saliva DNA for Next Generation Sequencing

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality DNA. However, DNA extraction from saliva can yield high quality DNA with little to no degradation/fragmentation that is suitable for a variety of DNA assays without the expense of a phlebotomist and can even be acquired through the mail. However, at present, no saliva DNA collection/extraction protocols for next generation sequencing have been presented in the literature. This protocol optimizes parameters of saliva collection/storage and DNA extraction to be of sufficient quality and quantity for DNA assays with the highest standards, including microarray genotyping and next generation sequencing.

DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.

收集和唾液DNA提取用于新一代测序

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality ...

Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina.

The use of cytobrush sampling to collect lymphocytes and monocytes from the endocervix is a minimally invasive technique that provides samples for analysis of female genital tract immunity. In this protocol, we describe the collection of cytobrush samples and immune cell isolation for flow cytometry assays.

收集，分离，和人类宫颈样品的流式细胞仪分析

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female ...

A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the patients are unaware of such disorders or are having difficulty expressing their symptoms. Until now, several devices for perspiration monitoring have been developed; however, such devices tend to have a relatively large exterior, considerable power consumption, and/or less sensitivity.
Recently, we developed a small, wireless device for perspiration monitoring. The device consists of a temperature/relative humidity (T/RH) sensor, battery-driven small data logger, and silica gel as a desiccant in a small cylindrical exterior. The T/RH sensor is placed between the detection windows (through which the water vapor from the skin enters) and the silica gel. The underlying principle of the perspiration monitoring device is based on Fick&#39;s law of diffusion, which means that water vapor flux from the skin to the silica gel (i.e.&#160;transepidermal water loss and perspiration) can be captured by change in humidity at the T/RH sensor. In addition, a baseline subtraction method was adopted to distinguish perspiration and transepidermal water loss.
As shown in the previous report, the developed device can monitor the perspiration at any sites of the body in an easy, wireless manner. However, detailed methods of how to use the device have not been disclosed yet. In this article, therefore, we would like to show the point-by-point tutorials of how to use the device for perspiration monitoring, by showing the sympathetic activity test with the sympathetic skin response monitoring as an example.

Recently, we developed a small wireless device for perspiration monitoring. In this article, we present detailed protocols on how to use the device for perspiration monitoring with an example of the sympathetic activity test.

详细的汗渍监控协议使用一种新的，小的，无线设备

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

解剖的小鼠胰腺的组织学分析与代谢谱

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Robotic Lateral Pancreaticojejunostomy for Chronic Pancreatitis

Lateral pancreaticojejunostomy (LPJ) has shown good postoperative outcomes in patients with painful, morphine dependent, chronic pancreatitis (CP). The recent rise of robotic and laparoscopic pancreatic surgery has found benefits such as reduced time to functional recovery. Few studies have reported on the feasibility, technique and outcome of robotic LPJ, especially including transection of the gastroduodenal artery. The present study describes the main steps for robotic LPJ in a patient with painful chronic pancreatitis with a dilated main pancreatic duct. The patient underwent robotic LPJ. The LPJ anastomosis is performed using a running suture technique in a longitudinal side-to-side manner. Routinely, the gastroduodenal artery is transected to drain the entire length of the main pancreatic duct. The patient is in French position; 7 trocars are placed (4 robotic, 2 laparoscopic assistants, 1 liver-retractor). After docking of the robot system, the omental bursa is opened, and the right gastroepiploic artery and vein are ligated at their base at the lower border of the pancreas. Intraoperative ultrasonography is performed in order to determine trajectory of the dilated main pancreatic duct which is opened for its entire length after the gastroduodenal artery has been suture ligated both cranially and caudally from the main pancreatic duct. A Roux limb is created, and a latero-lateral PJ is fashioned using several 3-0 barbed sutures. A stapled jejuno-jejunostomy is created at sufficient distance from the pancreatic anastomosis, aided by a 50 cm suture. The described technique for robotic LPJ is a complex but feasible operation for patients with treatment refractory CP and a dilated main pancreatic duct. Due to its complexity, implementation in high volume centers with extensive experience with CP surgery may improve outcomes.

Robotic lateral pancreaticojejunostomy (RLPJ) may be used in patients with painful, morphine dependent, chronic pancreatitis and a dilated main pancreatic duct. We describe a standardized and reproducible technique for RLPJ, which includes transection of the gastroduodenal artery.

慢性胰腺炎的机器人侧胰腺肌切除术

Lateral pancreaticojejunostomy (LPJ) has shown good postoperative outcomes in patients with painful, morphine dependent, chronic pancreatitis (CP). The ...

Laparoscopic Radical Left Pancreatectomy for Pancreatic Cancer: Surgical Strategy and Technique Video

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological resection of left-sided pancreatic cancer. Several surgical techniques have been described in recent years, but few were specifically designed for minimally invasive approaches. This study describes and demonstrates a standardized and reproducible technique for an adequate oncological resection of pancreatic cancer: laparoscopic radical left pancreatectomy (LRLP).
A 61-year-old woman presented with an incidental finding of a 3 cm mass in the left pancreas suspect for malignancy. Imaging did not reveal distant metastases, central vascular involvement, or morbid obesity, hence the patient was suitable for LRLP. This study describes the main steps of LRLP for pancreatic cancer. First, the lesser sac is opened by transecting the gastrocolic ligament. The splenic flexure of the colon is mobilized and the inferior border of the pancreas including Gerota&#39;s fascia is dissected down to the inferior border of the spleen. The pancreas is tunneled and hung, including Gerota&#8217;s fascia with a vessel loop. At the pancreatic neck, a tunnel is created between the pancreas and the portal vein, likewise a vessel loop is passed. The pancreas is then transected using the graded compression technique with an endostapler. Both the splenic vein and artery are transected before completing the resection. The entire specimen is extracted in a retrieval bag via a small Pfannenstiel incision.
Duration of the surgery was 210 min with 250 mL blood loss. Pathology revealed a R0-resection (&#62;1 mm) of a well-to-moderately differentiated adenocarcinoma originating from an intraductal papillary mucinous neoplasm. A total of 15 tumor-negative lymph nodes were resected. This is a detailed description of LRLP for left-sided pancreatic cancer as is currently being used within the international, multicenter randomized DIPLOMA (Distal Pancreatectomy Minimally Invasive or Open for PDAC) trial.

Oncologically safe left pancreatectomy requires radical resection (R0), Gerota&#8217;s (perirenal) fascia resection, and adequate lymph node dissection. This study describes the technical details of laparoscopic radical left pancreatectomy (LRLP), used in the first international multicenter randomized trial comparing minimally invasive with open left pancreatectomy for pancreatic cancer, the DIPLOMA trial.

胰腺癌的腹腔镜根左造影切除术:手术策略与技术视频

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological ...

Virtual Reality Tools for Assessing Unilateral Spatial Neglect: A Novel Opportunity for Data Collection

Unilateral spatial neglect (USN) is a syndrome characterized by inattention to or inaction in one side of space and affects between 23-46% of acute stroke survivors. The diagnosis and characterization of these symptoms in individual patients can be challenging and often requires skilled clinical staff. Virtual reality (VR) presents an opportunity to develop novel assessment tools for patients with USN.
We aimed to design and build a VR tool to detect and characterize subtle USN symptoms, and to test the tool on subjects treated with inhibitory repetitive transcranial magnetic stimulation (TMS) of cortical regions associated with USN.
We created three experimental conditions by applying TMS to two distinct regions of cortex associated with visuospatial processing- the superior temporal gyrus (STG) and the supramarginal gyrus (SMG) - and applied sham TMS as a control. We then placed subjects in a virtual reality environment in which they were asked to identify the flowers with lateral asymmetries of flowers distributed across bushes in both hemispaces, with dynamic difficulty adjustment based on each subject's performance. 
We found significant differences in average head yaw between subjects stimulated at the STG and those stimulated at the SMG and marginally significant effects in the average visual axis.
VR technology is becoming more accessible, affordable, and robust, presenting an exciting opportunity to create useful and novel game-like tools. In conjunction with TMS, these tools could be used to study specific, isolated, artificial neurological deficits in healthy subjects, informing the creation of VR-based diagnostic tools for patients with deficits due to acquired brain injury. This study is the first to our knowledge in which artificially generated USN symptoms have been evaluated with a VR task.

The goal was to design, build, and pilot a novel virtual reality task to detect and characterize unilateral spatial neglect, a syndrome affecting 23-46% of acute stroke survivors, expanding the role of virtual reality in the study and management of neurologic disease.

用于评估单边空间忽视的虚拟现实工具:数据收集的新机遇

Unilateral spatial neglect (USN) is a syndrome characterized by inattention to or inaction in one side of space and affects between 23-46% of acute stroke ...

Robot-Assisted Kidney Transplantation

This paper describes robot-assisted kidney transplantation (RAKT) from a living donor. The robot is docked between the parted legs of the patient, placed in the supine Trendelenburg position. Kidney allografts are provided by a living donor. Before vascular anastomosis, the kidney allograft is prepared by inserting a double-J stent in the ureter, and the temperature for the anastomosis is lowered by wrapping it in an ice-packed gauze. A 12 mm or 8 mm port for the robotic camera and three 8 mm ports for robotic arms are placed. A peritoneal pouch is created for the kidney allograft by raising the peritoneal flaps on both sides over the psoas muscle before dissecting the iliac vessels and bladder. A 6 cm Pfannenstiel incision is made to insert the kidney into the peritoneal pouch, lateral to the right iliac vessels. 
After clamping the external iliac vein with Bulldogs clamps, a venotomy is performed, and the graft renal vein is anastomosed to the external iliac vein in an end-to-side continuous manner with a 6/0 polytetrafluoroethylene suture. After clamping the graft renal vein, the iliac vein is declamped. This is followed by clamping of the external iliac artery, arteriotomy, arterial anastomosis with a 6/0 polytetrafluoroethylene suture, clamping of the graft renal artery, and declamping of the external iliac artery. Reperfusion is then carried out, and ureteroneocystostomy is performed using the Lich-Gregoir technique. The peritoneum is closed at a few locations with polymer locking clips, and a closed-suction drain is placed through one of the working ports. After deflating the pneumoperitoneum, all incisions are closed.

This paper provides technical details for robot-assisted kidney transplantation from a living donor.

机器人辅助肾移植

This paper describes robot-assisted kidney transplantation (RAKT) from a living donor. The robot is docked between the parted legs of the patient, placed ...