作者感谢李潘文凯针, 头 & 资深顾问, 儿童急诊, KK 妇女和儿童医院, 100 武吉天然路, 新加坡为行政支持与视频文章制作。在 "儿科紧急情况下金属异物摄取手持金属探测器" 许可下转载的文章;2018年8月:85 (8): 618–624;Hazwani Binte 哈姆扎王子、守夜詹姆斯、苏拉吉马尼卡姆、Sashikumar Ganapathy: 版权所有印度儿科杂志。

该议定书遵循机构人类研究伦理委员会的准则。中央机构审查委员会为这项研究提供了道德上的批准。
1. 参加者筛选
<ol><li>招募向儿科急诊科的病人提供异物摄取病史。</li>
	<li>排除呼吸窘迫的病人, 并植入了类似于速度器的设备。</li>
</ol>2. 准备病人
<ol><li>给病人穿上医院礼服。</li>
	<li>删除所有可能包含金属物体, 如衣服, 眼睛眼镜, 耳环, 挂件, 和饰物在检查之前, 以避免干扰 HHMD 筛查。</li>
	<li>把病人放在房间的中心, 远离其他潜在的金属干扰。</li>
</ol>3. 准备考官和照顾者
<ol><li>卸下所有金属配件, 如首饰和手表。</li>
	<li>请护理人员删除所有金属配件, 如首饰和手表。</li>
</ol>4. 金属探测器筛查
<ol><li>使用 HHMD (图 1) 进行筛选。</li>
	<li>通过在金属片上挥舞一个正面的视听信号, 确认 HHMD 的工作状态。</li>
	<li>对较大的孩子进行 HHMD 扫描。</li>
	<li>在婴儿的直立位置进行 HHMD 扫描, 让看护者适当地控制病人。</li>
	<li>握住病人的手, 远离身体。</li>
	<li>将病人的头部放在延伸处以暴露颈部。</li>
	<li>从下巴的水平扫描到髋关节的水平, 从颈部到骨盆覆盖解剖区域。</li>
	<li>在整个身体内以曲折的方式执行扫描, 以确保扫描每个区域。</li>
	<li>扫描前方和向后作为患者的厚度必须考虑。</li>
	<li>仔细注意在扫描期间的正面视听信号 MFBs 存在。</li>
</ol>5. 执行相关的射线照相
<ol><li>根据 MFBs HHMD 筛查的区域, 订购颈部、胸部或腹部的相关射线。</li>
	<li>如果在 HHMD 筛查时没有发现 MFB, 则继续进行所有 X 线观察 (颈部、胸部和腹部), 而对异物摄取的怀疑的临床指标很高。</li>
</ol>6. 记录测试结果
<ol><li>记录根据工作流专门创建的日志 (图 2) 中的测试结果 (图1)。</li>
	<li>通过 palpating 以下突出的骨性地标-胸骨凹槽、xiphisternum、耻骨联合和双侧髂嵴, 将正的视听 HHMD 信号与身体底层的骨性地标相关联。</li>
	<li>在进行 X 光检查之前, 在航海日志的解剖参考提纲上记录地标。</li>
</ol>

<ol>
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作者没有什么可透露的。

金属异物在儿童中摄取是一个常见的问题, 在大多数患者中, 这是保守的管理7,16。单一或重复的射线照相可以暴露敏感的儿科人群的不必要的辐射风险, 而试图本地化的摄取金属异物。一些研究已经描述了使用金属探测器在 MFB 摄取儿童, 灵敏度超过 96%, 特异性大于 80%1,7,17,<sup...

由于婴儿和儿童的自然好奇性, 通过品尝和吞咽1,2来探索物体, 异物摄取在儿童急诊室中是一个普遍存在的问题。以前的研究报告, 金属物体构成85% 的所有异物 ingestions 在他们的队列的病人3,4。本议定书的目标是评估使用手持金属探测器检测儿童摄取的金属异物的准确性和可行性。我们建议, 通过在对金属异物摄取高度怀疑的儿童的分流过程中及早引入手持金属检测器筛选协议, 被下令定位金属异物的射线照相数量可以减少了这种对无线电敏感的人群。
手持金属探测器 (HHMD) 是一种价格低廉、易于获取的辅助辅料, 可帮助加快对摄取金属异物的管理。金属探测器是一种非常低频的装置, 它有一个接收器和一个发射机, 它能检测到在其附近3的金属物体的存在所引起的电磁场的变化。如果金属物体在磁场中, 电磁场就会中断, 传感器将在设备4上设置视听报警。金属探测器检查可以在不到2分5分钟内彻底完成。因此, HHMD 筛查可以在早期的诊断儿童的分流过程中高怀疑金属异物摄取的地形定位。
硬币是最常见的吞食金属异物1,4,6,7。由于缺乏锋利的边缘和一般无毒的性质, 大多数硬币 ingestions 管理保守。一般摄取的其他类型的材料包括磁铁、纽扣电池、小玩具、花生、首饰、纽扣、骨头、种子和爆米花8、9。异物的冲击最常见发生在食管三处, 即上食管括约肌, 中食管在主动脉弓水平, 下食管括约肌10。金属异物 (MFBs) 位于食管可能是无症状的40% 患者11,12 , 其长期的撞击可能导致各种并发症, 如食管穿孔, 纵隔,气管食管瘘, 气管狭窄, 主动脉食管瘘1,13,14,15。其他部位的异物嵌塞包括幽门, 十二指肠和 ileo-盲肠交界10。
执行颈部、胸部和腹部的射线照相来诊断疑似异物摄取是5紧急部门的普遍做法。在所有被摄取的异物中, 只有10% 是无线电不透明的10。放射检查是耗时的, 并有危险的辐射暴露在儿童。精确的地形定位的摄取金属物体是非常重要的指导治疗。在分流过程早期引入 HHMD 筛查可以通过地形对摄取的金属异物进行定位, 帮助减轻放射摄影的外部排列。
本研究评价了 HHMD 在系统地形学中定位 MFBs 的准确性, 确定了 HHMD 在当地儿科人群中的有效性和可行性。我们还研究了这种辐射免费筛查工具是如何良好的病人和家长接受。在可能和2016年7月之间, 所有连续的患者递交到儿科急诊科以外国身体摄取的历史被包括在研究中。知情同意是从父母或法定监护人那里获得的。使用 HHMD 对指称异物摄取的患者进行系统扫描。对摄取金属异物进行鉴定和定位的金标准是对宫颈区、胸腔和腹部的普通影像。为便于进行研究, 创建了一个标准化的工作流 (图 1)。根据工作流专门创建的日志簿用于收集初始数据, 随后将其转录到电子表格 (图 2)。结果部分中的数据被描述为按百分比显示的事例数。

<table>
	<tbody>
		<tr>
			<td>Hand held metal detector</td>
			<td>Garrett Super Scanner V</td>
			<td>NA</td>
			<td>NA</td>
		</tr>
	</tbody>
</table>

handheld metal detector screening for metallic foreign body ingestion in children

硬币是儿童中最常见的摄取金属异物。本议定书的目标是评估使用手持金属探测器检测儿童摄取的金属异物的准确性和可行性。我们建议, 通过引进手持金属探测器筛查早期的儿童的分流过程中, 高怀疑金属异物摄取, 可以减少的射线照相的数量, 以本地化金属异物可在此无线电敏感的人口。这项研究的议定书要求对参加者进行检查, 以了解外国身体摄取和排除呼吸窘迫或金属植入物患者的病史。病人改变了医院礼服和物品, 可能含有金属如眼镜, 耳环, 挂件, 和装饰品被删除。病人被安置在房间的中心远离其他金属干扰。手持式金属探测器的工作状态首先通过引出一个正面的视听信号来确认。然后, 筛查是在一个直立的位置, 头部延伸, 以揭露颈部, 从下巴水平到髋关节水平, 以前方和向后的曲折方式覆盖解剖区域从颈部到骨盆。在扫描时仔细注意到有金属异物存在的正面视听信号。有关的射线照相根据检测到的金属探测器筛查的区域排序。手持金属探测器能够精确地识别我们研究中摄取的金属异物中的所有硬币。手持式金属探测器不能始终如一地检测非硬币金属异物。该协议说明手持金属探测器在识别和定位硬币和硬币等金属异物方面的准确性。

研究人员为这项观察性研究招募了36名病人。这些病人已向儿科急诊科提交了一份外国身体摄取的病史。其中28例患者有金属异物 ingestions (图 3)。其余的8例患者摄取了非金属异物, 如鱼骨、沙爹棒和塑料纽扣。硬币是最常见的异物摄取类型 (表 1), 其中包括30.5% 的病人。其他最常见的异物, 被摄取包括鱼骨, 玩具零件, 纽扣电池, 金属螺丝。...

Watch this Scientific Journal Video about Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children at JoVE.com

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

在这里, 我们提出了一个使用手持金属探测器的协议, 用来筛查那些向儿科急诊医学系提供异物摄取病史的儿童的摄入金属异物。

金属异物在儿童体内摄取的手持金属探测器筛选

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a ...

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10.6K 次观看.  KK women's and Children's Hospital. 在这里, 我们提出了一个使用手持金属探测器的协议, 用来筛查那些向儿科急诊医学系提供异物摄取病史的儿童的摄入金属异物。

视频: Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

High Content Screening in Neurodegenerative Diseases

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be addressed as a matter of great urgency1-4. Multiple complementary 'omics' approaches have provided clues as to the genetic risk factors and pathogenic mechanisms underlying numerous neurodegenerative diseases, but most findings still require functional validation5. For example, a recent genome wide association study for Parkinson's Disease (PD), identified many new loci as risk factors for the disease, but the underlying causative variant(s) or pathogenic mechanism is not known6, 7. As each associated region can contain several genes, the functional evaluation of each of the genes on phenotypes associated with the disease, using traditional cell biology techniques would take too long. 
	There is also a need to understand the molecular networks that link genetic mutations to the phenotypes they cause. It is expected that disease phenotypes are the result of multiple interactions that have been disrupted. Reconstruction of these networks using traditional molecular methods would be time consuming. Moreover, network predictions from independent studies of individual components, the reductionism approach, will probably underestimate the network complexity8. This underestimation could, in part, explain the low success rate of drug approval due to undesirable or toxic side effects. Gaining a network perspective of disease related pathways using HT/HC cellular screening approaches, and identifying key nodes within these pathways, could lead to the identification of targets that are more suited for therapeutic intervention.
	High-throughput screening (HTS) is an ideal methodology to address these issues9-12. but traditional methods were one dimensional whole-well cell assays, that used simplistic readouts for complex biological processes. They were unable to simultaneously quantify the many phenotypes observed in neurodegenerative diseases such as axonal transport deficits or alterations in morphology properties13, 14. This approach could not be used to investigate the dynamic nature of cellular processes or pathogenic events that occur in a subset of cells. To quantify such features one has to move to multi-dimensional phenotypes termed high-content screening (HCS)4, 15-17. HCS is the cell-based quantification of several processes simultaneously, which provides a more detailed representation of the cellular response to various perturbations compared to HTS. 
	HCS has many advantages over HTS18, 19, but conducting a high-throughput (HT)-high-content (HC) screen in neuronal models is problematic due to high cost, environmental variation and human error. In order to detect cellular responses on a 'phenomics' scale using HC imaging one has to reduce variation and error, while increasing sensitivity and reproducibility. 
	Herein we describe a method to accurately and reliably conduct shRNA screens using automated cell culturing20 and HC imaging in neuronal cellular models. We describe how we have used this methodology to identify modulators for one particular protein, DJ1, which when mutated causes autosomal recessive parkinsonism21. 
	Combining the versatility of HC imaging with HT methods, it is possible to accurately quantify a plethora of phenotypes. This could subsequently be utilized to advance our understanding of the genome, the pathways involved in disease pathogenesis as well as identify potential therapeutic targets.

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

在神经退行性疾病的高内涵筛选

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be ...

科研

医学

Eye Tracking Young Children with Autism

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a direct, detailed and objective measure of gaze behavior, eye-tracking has become a valuable tool for examining abnormal perceptual strategies in clinical populations and has been used to identify disorder-specific characteristics1, promote early identification2, and inform treatment3. In particular, investigators of autism spectrum disorders (ASD) have benefited from integrating eye-tracking into their research paradigms4-7. Eye-tracking has largely been used in these studies to reveal mechanisms underlying impaired task performance8 and abnormal brain functioning9, particularly during the processing of social information1,10-11. While older children and adults with ASD comprise the preponderance of research in this area, eye-tracking may be especially useful for studying young children with the disorder as it offers a non-invasive tool for assessing and quantifying early-emerging developmental abnormalities2,12-13. Implementing eye-tracking with young children with ASD, however, is associated with a number of unique challenges, including issues with compliant behavior resulting from specific task demands and disorder-related psychosocial considerations. In this protocol, we detail methodological considerations for optimizing research design, data acquisition and psychometric analysis while eye-tracking young children with ASD. The provided recommendations are also designed to be more broadly applicable for eye-tracking children with other developmental disabilities. By offering guidelines for best practices in these areas based upon lessons derived from our own work, we hope to help other investigators make sound research design and analysis choices while avoiding common pitfalls that can compromise data acquisition while eye-tracking young children with ASD or other developmental difficulties.

Eye tracking has long been used to study gaze patterns in typically-developing individuals, but recent technological advancements have made its use with clinical populations, including autism, more feasible. While eye-tracking young children with autism can offer insight into early symptom manifestations, it involves methodological challenges. Suggestions for best practices are provided.

自闭症幼儿的眼动追踪

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

持续时间较长的小鼠全身热疗协议

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory rate, salivation, perspiration and renal function. The ANS has two main branches: the sympathetic nervous system, preparing the human body for action in times of danger and stress, and the parasympathetic nervous system, which regulates the resting state of the body.
ANS activity can be measured invasively, for instance by radiotracer techniques or microelectrode recording from superficial nerves, or it can be measured non-invasively by using changes in an organ's response as a proxy for changes in ANS activity, for instance of the sweat glands or the heart. Invasive measurements have the highest validity but are very poorly feasible in large scale samples where non-invasive measures are the preferred approach. Autonomic effects on the heart can be reliably quantified by the recording of the electrocardiogram (ECG) in combination with the impedance cardiogram (ICG), which reflects the changes in thorax impedance in response to respiration and the ejection of blood from the ventricle into the aorta. From the respiration and ECG signals, respiratory sinus arrhythmia can be extracted as a measure of cardiac parasympathetic control. From the ECG and the left ventricular ejection signals, the preejection period can be extracted as a measure of cardiac sympathetic control. ECG and ICG recording is mostly done in laboratory settings. However, having the subjects report to a laboratory greatly reduces ecological validity, is not always doable in large scale epidemiological studies, and can be intimidating for young children. An ambulatory device for ECG and ICG simultaneously resolves these three problems.
Here, we present a study design for a minimally invasive and rapid assessment of cardiac autonomic control in children, using a validated ambulatory device 1-5, the VU University Ambulatory Monitoring System (VU-AMS, Amsterdam, the Netherlands,<a href="http://www.vu-ams.nl" target="_blank"> www.vu-ams.nl</a>).

Measuring Cardiac Autonomic Nervous System ANS Activity in Children

Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children.

测量心脏自主神经系统（ANS）儿童活动

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

对黑色素瘤修饰符一个斑马鱼原地肿瘤模型的筛选

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

一种新型的高分辨率<em&gt;在体内</em&gt;成像技术研究肿瘤生长及治疗颅内结构的动态响应

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

一种新型的<em&gt;在体内</em&gt;基因转移技术和<em&gt;在体外</em&gt;细胞分析为肌肉骨骼疾病骨质流失的研究

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

常温<em&gt;离体</em&gt;肾脏的灌注肾的移植保护移植前

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

异位自体肾移植的猪模型:有步骤，分步协议

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

在女小鼠害保留神经的中期尿道梗阻（NEMO）

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...