特别感谢莎拉Kossak酒店，米切尔Guanzon，宋允杨和李郭嘉，为他们的实验动物的内镜检查过程中他们的合作。

1。设立内窥镜系统<ol><li>设置了内窥镜系统麻醉的小鼠前，按照制造商的说明进行操作。使用视频输尿管镜专为人类获得本协议（ 图1）。插入管的外径是9.9，FR（3.3毫米），其远端是8.5，FR（2.8毫米）和仪器通道的内径为6周五（1.2毫米）。 </li><li>虽然使用内窥镜尚未链接到疾病传播[9]，消毒的内窥镜和透镜与孢子形成微生物工作时，用70％乙醇10或者用适当的技术。准备抑菌消毒润滑剂和非刮拭组织饱和内窥镜末端。 </li><li>附上一个3毫升注射器充满空气（或合适的CO 2气体的混合物）的仪器通道的顶部。 </li><li>打开电源，并设置白平衡正常的白光，以及窄带成像（NBI）模式下，当适用。自2001年以来，NBI提供诊断成像的好处，因为它强调精细的血管结构上粘膜，包括溃疡和肿瘤病变，其中有特征的异常血管11。 NBI过滤模式限制捕获的光的波长为390-445 nm和530-550 nm处，其中突出的结构与血红蛋白。 </li><li>打印考核评分表（ 图2）来记录你的发现。熟悉预期的正常和NBI图像（ 图3A）和定义，以及每项准则的成绩在考试前（见第6.1节）。 </li><li>准备在磷酸盐缓冲盐水（PBS）中的着色剂为基础的解决方案，色素​​，如果需要的话。在色素，使用染料作为造影剂，以突出黏膜变化进行了考试。用亚甲基蓝（0.1-1％）和靛蓝胭脂红（0.1％）解决方案（ 图3B）。注该亚甲蓝是一种氧化还原剂可以制作氧化自由基，间接交联氨基酸残基，和DNA降解12，这可能会干扰样品的分析。 </li></ol> 2。鼠标的全身麻醉<ol><li>选择麻醉剂和给药途径。从有关药物与免疫系统和研究目标的潜在干扰动物资源的设施或机构动物护理和使用委员会（IACUC）请教兽医。有些药物可能会影响细胞因子的表达分布在动物，包括啮齿动物13。使用注射三溴乙醇的终端程序或异氟醚生存内镜，以及适当的控制。 </li><li>用于感应，辖异氟醚4-5％在100％的氧气以0.2-0.5升/分钟的速率;使用0.5-2％，进行维护。这个协议是不适合于所有的动物。请咨询您的兽医机构进行适当的指导和培训。通过有关建议IACUC遵守，以实现动物福利的最高标准。确保鼠标，如果完全麻醉。 </li><li> &#39;带&#39;麻醉鼠标在使用实验室胶带背部或腹部位置的支撑面表。当心你的内窥镜屏幕上的解剖定位。确保其下肢和尾巴。磁带的带也可放置在胸前，以防止运动远离呼吸器，但要避免限制胸廓呼吸运动。 </li></ol> 3。详细内镜检查<ol><li>寻找肛周病变插入内窥镜伸入直肠前，然后经肛门推进范围。 </li><li>大衣与无菌手术润滑剂内窥镜。千万不要涂在CCD（电荷耦合器件）传感器的表面。 </li><li>插入内窥镜，并开始录制。录制的视频是米有用内镜检查，如果需要检查后eticulous评价。尽量在时间，以减少肠/粘膜有刺激性尽可能少的长度进行胃镜检查。 </li><li>通过使用注射器慢慢地分开设置肠壁施加空气。根据鼠标的大小，使用1-2毫升的空气。避免过多的空气吹入，它可能会导致麻醉小鼠的呼吸窘迫和死亡。 </li><li>推进内窥镜前进你开道用空气或范围的轻柔旋转。没有前进的内窥镜，如果你没有看到肠腔。穿孔或直肠/结肠眼泪可以发生在这个阶段，尤其是在年轻小鼠（&lt;8-10周龄）。拍照是必要的。 </li><li>随着范围的进步，评估是否有自发性出血（定义为自然发生粘膜出血与外伤性内镜相关的）。自发出血可作为内窥镜进入直肠和结肠被观察到。 </li><li>如SESS结肠壁的透明度。透明度定义为可视化的壁间血管在结肠和其他周围的内脏（ 图4）的能力。范围温柔的动作（向后转，顺时针，逆时针）将有助于确定血管可以在其他脏器的角度来看待。肠系膜脂肪组织也可见到。注意如果盆腔内直肠似乎比结肠壁在腹腔透明度较低。 </li><li>检查，以确定病灶（水肿区，糜烂，溃疡，肿瘤质量）的类型和数量。这将使用结肠炎得分，并已开发出的小数标识符被记录在计分的形式。 </li><li>一旦其中结肠尾端弯曲成近端结肠的面积达到了，撤内窥镜慢慢寻找接触性出血（的发生是由于后粘膜脆性的存在内窥镜前进或删除）。它没有必要故意刮结肠表面。经过适当的内镜检查健康的大肠黏膜不流血。 </li></ol> 4。活检<ol><li>当需要活检，通过专用仪器内窥镜通道插入活检钳。监控内窥镜屏幕上的镊子的进步，以防止肠穿孔。 </li><li>打开钳活检杯到约60-80℃杯对杯角，并指导他们感兴趣的区域。过度打开钳子或压力可以把握肠组织，这会导致健康透明肠壁活检引起的穿孔的额外层。如果发生穿孔，麻醉期间安乐死的动物按照你IACUC建议。穿孔导致内毒素血症和腹膜炎。 </li><li>关闭活检杯，同时保持钳闭拉出。 </li><li>圣雷莫通过冲洗组织一个解决方案，是温和的组织和钳子已经从钳子试样。后来的RNA和PBS是共同的工作方案，如果需要，使用针撞出样品。避免侵蚀着钳子杯或切削刃。 </li></ol> 5。麻醉恢复期<ol><li>待考试结束，提供一个温馨的环境，鼠标，直到它恢复。使用任一种电光源，或温暖垫定在37°C避免过热;周围血管扩张危及动物的恢复。密切监视鼠标，直到完全康复。 </li></ol> 6。大肠健康与十进制标识符综合内镜评分系统<ol><li>使用评分表（ 图2）及以下的参数定义（ 小鼠大肠炎症和肿瘤的内镜评估使用十进制加权计分法：参数定义和协议 ）实例和可选的&#39;远端近端结肠炎内镜评分系统“（D-PECS），作为无偏差评估和详细记录保存的指引。 </li><li>分数的计分制的形式（ 图2）中所述的四个参数的大肠炎症。精心评估肛周区域，肠透明度，肠出血的类型（如果存在），以及存在和病灶的数量（糜烂，溃疡和/或肿瘤细胞）。 </li><li>在评估肿瘤为研究目的，研究的数量，位置和大小，并使用结肠图和计分表上条形图描述它们。得到的分数和病变的数目（当适用时）的数值总和将确定背景结肠的炎症，最主导型病变，以及病变的数目。 </li></ol>

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没有财务或披露利益冲突。

有关于修改内窥镜技术在小鼠和故障排除几方面的考虑。需要使用灵活的内镜检查过程中掌握了过程的关键方面包括需要防止腹胀和鼠标的呼吸妥协的风量的调节，以及视图控件的细微的技术方面与协调的角度和转矩控制。转矩控制是很重要的，以减少肠穿孔的危险。如果处理不当，鼠灵活的内窥镜成为一个非常安全的，快速的程序，可以直接可视化和感兴趣的研究领域肠活检取样。 <p class="j...

鼠内镜检查已经完成作为一个有用的研究工具超过十年1-3。到目前为止，大多数研究采用小鼠内窥镜使用了硬性内窥镜，虽然有些也使用软式乙状结肠镜。鼠胃镜提供立竿见影的效果和肠道正常，炎症的严重程度和肿瘤进展比较间接的措施，比如形体消瘦，腹泻，和组织学（这是次优的，当病灶呈斑片状）的程度更客观的估计数字，而提供对结肠的整体健康状况的见解。最值得注意的是，内窥镜允许的实验室动物模型随时间的反复评价，相对于传统的组织学检查，需要被实施安乐死，并收获用于分析1冒号的动物。尽管有这些优点，利用鼠内窥镜技术还没有广泛和由缺乏implementati标准化协议的限制在和病理结果的得分。正确实施，鼠胃镜拥有巨大潜力，以进一步促进我们的理解和多种慢性胃肠疾病，包括炎症性肠道疾病，结肠炎，大肠肿瘤动物模型的表征。 小鼠内镜技术的效用有赖于其可重复性和客观性，这就需要标准化的审查程序的存在和一致的，非冗余，可靠的评分系统肠道病变。至少三个描述性的评分系统对结肠炎4-6的内窥镜的评估，以及用于结肠肿瘤的小鼠4,7,8已经出版。然而，这些报道办法和评分系统是不能跨的研究容易比较。在许多情况下，有一种用于病变的分类标准没有明确的定义，和标准给出时，它们有很大的不同。此外，没有报告的评分系统是整合双方结肠炎和肿瘤，二是可以同时发生，对结果的互动效应最常见的结肠病理，到一个单一的测量工具的评估。最后，确实存在对结肠炎的内镜评分系统往往有炎症个类别，有限制的歧视（ 即 ，在短整型尺度窄的得分选择，往往1-4），以正确地表示疾病进展的情况，并能够使用参数统计分析。 在本文中，我们描述了使用柔性内窥镜的评估鼠结肠炎和结肠肿瘤的严重程度和描述的标准化协议，用于采用该技术的肛周区域，直肠和结肠的解剖评估。我们在内窥镜说明有效的故障排除，以减少创伤和图像伪影，和我们描述了一个可重复的评分系统基于验证的已发表的临床参数。该评分系统集成使用的有12个可能的等级炎症，肿瘤测绘和绘图选项，以及一个新的十进制单位制（即小数点标识符）歧视高水平突出与特定的诊断价值发现两者肠道炎症和肿瘤的评价（ 即肿瘤的发展，内镜检查等过程中的并发症）。使用十进制标识符允许对相关的数据集进行进一步的下游分析的快速跟踪。最后，我们在肠炎和结肠炎相关癌（葡聚糖硫酸钠（DSS）诱导的结肠炎， 难辨梭状芽孢杆菌感染，氧化偶氮甲烷/ DSS诱导结肠癌）的多种小鼠模型进行可靠性和评分系统的有效性测试。

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			<td height="21" style="height:21px;width:240px;">Isoflurane, USP</td>
			<td style="width:160px;">Webster Veterinary&#160;</td>
			<td style="width:217px;"></td>
			<td style="width:79px;"></td>
		</tr>
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			<td height="20" style="height:20px;width:240px;">Surgical lubricant</td>
			<td style="width:160px;">Savage laboratories</td>
			<td style="width:217px;">surgilube, 0281-0205-45</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:240px;">Phosphate Buffered Saline</td>
			<td style="width:160px;">Thermo Scientific&#160;</td>
			<td style="width:217px;">SH30256</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:240px;">RNAlater</td>
			<td style="width:160px;">Ambion</td>
			<td style="width:217px;">AM7021</td>
			<td style="width:79px;"></td>
		</tr>
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			<td height="20" style="height:20px;width:240px;">Methylene Blue 1% Aqueous Solution</td>
			<td style="width:160px;">Fisher Science Education</td>
			<td style="width:217px;">S96393</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:240px;"></td>
			<td style="width:160px;"></td>
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			<td height="21" style="height:21px;width:240px;">Flexible digital ureteroscope</td>
			<td style="width:160px;">Olympus America</td>
			<td style="width:217px;">URF-V</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Video system center</td>
			<td style="width:160px;">Olympus America</td>
			<td style="width:217px;">VISERA Pro OTV-S7 Pro</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Xenon light source</td>
			<td style="width:160px;">Olympus America</td>
			<td style="width:217px;">VISERA Pro CLV-S40 Pro</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Video recorder</td>
			<td style="width:160px;">MediCapture, Inc.</td>
			<td style="width:217px;">MediCap USB200</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Flexible biopsy cup forceps</td>
			<td style="width:160px;">Olympus America</td>
			<td style="width:217px;">FBC-3115</td>
			<td style="width:79px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Anesthesia machine</td>
			<td style="width:160px;">Euthanex Corporation</td>
			<td style="width:217px;">EZ-7000 Classic System</td>
			<td style="width:79px;"></td>
		</tr>
	</tbody>
</table>

flexible colonoscopy in mice to evaluate the severity of colitis and colorectal tumors using a validated endoscopic scoring system

为研究目的而利用现代内镜，大大方便了我们的胃肠道病变的认识。特别是，实验内窥镜一直需要在一个单独的实验动物反复评估，如那些评价慢性炎症性肠疾病的机制和结直肠癌的进展的研究是非常有用的。然而，对不同的研究中使用的方法是高度可变的。至少有三个内镜评分系统已经出版的小鼠结肠炎和公布结直肠肿瘤的评估，协议未能解决伴随结肠炎症的存在。本研究开发和验证一个可重复的内窥镜计分制度，同时集成了炎症和肿瘤的评价。这种新颖的评分系统由三大部分组成：1）评估大肠炎症的程度和严重性（基于肛周发现，沃尔玛的透明度升，粘膜出血，和局灶性病变），2）肿瘤性病变（格图和条形图），和临床病例通过病理和研究的相关基于十进制单位分配的类别观察病变3）数值排序的定量记录和内镜并发症（十进制标识符）。视频和手稿本文中所呈现的制备，以下IACUC批准的协议，以允许研究者使用经过验证和高度可重复的方法内窥镜得分自己的实验小鼠中，与该系统选项从近端内窥镜结肠炎（D-PECS区分远侧）。

该协议和图片在本研究中提出的制备用Olympus URF-V的柔性内窥镜14和NBI 11,15。视频图像被保存为MPEG-2格式和JPG，TIF，PNG或最大1280×1024像素，可转让的，以标准的USB闪存便携式记忆DICOM格式的图片。 NBI增强含血红蛋白的组织的视觉对比度。正常光线和NBI成像的关键特征，说明大肠黏膜的正常解剖及溃疡和肿瘤的发展（ 图3A和6A）在正常和异常血管之间的差异?...

Watch this Scientific Journal Video about Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System at JoVE.com

Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System

在过去的几年中，新一代内窥镜已成为重要的诊断艾滋病的研究评估小鼠结肠炎和结肠肿瘤。我们在此提出一个详细的协议，用于炎症和大肠肿瘤在小鼠体内的内镜检查，以及使用十进制标识符记录结肠炎和结肠直肠肿瘤的内镜严重性一种新的评分系统。

灵活的结肠镜检查在小鼠使用经过验证的内镜评分系统来评估结肠炎和结肠直肠肿瘤的严重程度

The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental ...

flexible-colonoscopy-mice-to-evaluate-severity-colitis-colorectal

Research

JoVE Journal

Medicine

31.5K 次观看.  Case Western Reserve University School of Medicine, Cleveland. 在过去的几年中，新一代内窥镜已成为重要的诊断艾滋病的研究评估小鼠结肠炎和结肠肿瘤。我们在此提出一个详细的协议，用于炎症和大肠肿瘤在小鼠体内的内镜检查，以及使用十进制标识符记录结肠炎和结肠直肠肿瘤的内镜严重性一种新的评分系统。 

视频: Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System

Establishment and Propagation of Human Retinoblastoma Tumors in Immune Deficient Mice

Culturing retinoblastoma tumor cells in defined stem cell media gives rise to primary tumorspheres that can be grown and maintained for only a limited time. These cultured tumorspheres may exhibit markedly different cellular phenotypes when compared to the original tumors. Demonstration that cultured cells have the capability of forming new tumors is important to ensure that cultured cells model the biology of the original tumor. 
Here we present a protocol for propagating human retinoblastoma tumors in vivo using Rag2-/- immune deficient mice. Cultured human retinoblastoma tumorspheres of low passage or cells obtained from freshly harvested human retinoblastoma tumors injected directly into the vitreous cavity of murine eyes form tumors within 2-4 weeks. These tumors can be harvested and either further passaged into murine eyes in vivo or grown as tumorspheres in vitro. Propagation has been successfully carried out for at least three passages thus establishing a continuing source of human retinoblastoma tissue for further experimentation. 
Wesley S. Bond and Lalita Wadhwa are co-first authors.

A method is described to propagate human retinoblastoma tumors in mice. Tumor cells are directly injected into the eyes of immune deficient mice. Secondary tumors have been successfully established using both cells directly harvested from human tumors and cultured tumorspheres.

在免疫缺陷小鼠的建立和传播人类视网膜母细胞瘤肿瘤

Culturing retinoblastoma tumor cells in defined stem cell media gives rise to primary tumorspheres that can be grown and maintained for only a limited ...

科研

医学

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

脓毒症动物模型，用于评估新型草药疗法

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

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 ...

Dry Powder and Nebulized Aerosol Inhalation of Pharmaceuticals Delivered to Mice Using a Nose-only Exposure System

Obstructive respiratory diseases like asthma and chronic obstructive pulmonary disease (COPD) are currently treated by inhaled anti-inflammatory and bronchodilator drugs. Despite the availability of multiple treatments, both diseases are growing public health concerns. The majority of asthma patients are well controlled on current inhaled therapies but a substantial number of patients with severe asthma are not. Asthma affects an estimated 300 million people worldwide and approximately 20 percent have a severe form of the disease. In contrast to asthma, there are few effective therapies for COPD. An estimated 10% of the population has COPD and the trend in death rates is increasing for COPD while decreasing for other major diseases. Although developing drugs for inhaled delivery is challenging, the nose-only inhalation unit enables direct delivery of novel drugs to the lung of rodents for pre-clinical efficacy and safety/toxicology studies. Inhaled drug delivery has multiple advantages for respiratory diseases, where high concentration in the lung improves efficacy and minimizes systemic side effects. Inhaled corticosteroids and bronchodilators benefit from these advantages and inhaled delivery may also hold potential for future biologic therapies. The inhalation unit described herein can generate, sample for characterization, and uniformly deposit a drug aerosol in the lungs of rodents. This enables the pre-clinical determination of the efficacy and safety of drug doses deposited in the lungs of rodents, key data required before initiating clinical development.

The inhalation unit described herein can generate, sample for characterization, and uniformly deposit a drug aerosol in the lungs of rodents. This enables the pre-clinical determination of the efficacy and safety of drug doses deposited in the lungs; key data enabling clinical inhaled drug development.

干粉和药品的雾化吸入气雾剂递送小鼠使用鼻只曝光系统

Obstructive respiratory diseases like asthma and chronic obstructive pulmonary disease (COPD) are currently treated by inhaled anti-inflammatory and ...

Taste Exam: A Brief and Validated Test

The emerging importance of taste in medicine and biomedical research, and new knowledge about its genetic underpinnings, has motivated us to supplement classic taste-testing methods in two ways. First, we explain how to do a brief assessment of the mouth, including the tongue, to ensure that taste papillae are present and to note evidence of relevant disease. Second, we draw on genetics to validate taste test data by comparing reports of perceived bitterness intensity and inborn receptor genotypes. Discordance between objective measures of genotype and subjective reports of taste experience can identify data collection errors, distracted subjects or those who have not understood or followed instructions. Our expectation is that fast and valid taste tests may persuade researchers and clinicians to assess taste regularly, making taste testing as common as testing for hearing and vision. Finally, because many tissues of the body express taste receptors, taste responses may provide a proxy for tissue sensitivity elsewhere in the body and, thereby, serve as a rapid, point-of-care test to guide diagnosis and a research tool to evaluate taste receptor protein function.

This protocol measures human taste responses and includes a brief anatomical assessment, a short taste test, and a validation method using the subject&#39;s reported sensation and taste receptor genotype.

味觉考试: 一个简短的验证测试

The emerging importance of taste in medicine and biomedical research, and new knowledge about its genetic underpinnings, has motivated us to supplement ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

小鼠颞叶癫痫的 Radiotelemetry 模型及脑电图监测

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ROM is called a joint contracture and results in increased morbidity. Due to the difficulty of reversing established knee contractures, early detection is important, and hence, knowing risk factors for their development is essential. The rat represents a good model with which the effect of an intervention can be studied due to the similarity of rat knee anatomy to that of humans, the rat's ability to tolerate long durations of knee immobilization in flexion, and because mechanical data can be correlated with histologic and biochemical analysis of knee tissue.
Using an automated arthrometer, we demonstrate a validated, precise, reproducible, user-independent method of measuring the extension ROM of the rat knee joint at specific torques. This arthrometer can be used to determine the effects of interventions on knee joint ROM in the rat.

The goal of the protocol is to measure the extension range of motion of the rat knee. The effects of various diseases that increase the stiffness of the knee joint and the effectiveness of treatments can be quantified.

用膝关节关节仪评价大鼠膝关节屈曲收缩的特定组织贡献

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ...

Murine Appendectomy Model of Chronic Colitis Associated Colorectal Cancer by Precise Localization of Caecal Patch

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal cancer, inflammatory bowel disease, and Parkinson&#8217;s disease. To study the function of the appendix, a gut disease-associated murine appendectomy model has been established and its step-by-step protocol is described here. This report introduces a facile protocol for caecal patch removal in mice followed by the chemical induction of chronic colitis-associated colorectal cancer using a combination of dextran sulfate sodium (DSS) and azoxymethane (AOM). IgA specific cells and IgA concentration were significantly reduced upon removal of the caecal patch in male C57BL/6 mice&#160;compared to those in the sham group. Simultaneously administering 2% DSS and AOM resulted in nearly 80% mice survival in both sham and appendectomy groups without significant body weight loss. Histological results confirmed colonic inflammation and different degrees of adenocarcinoma. This model can be used for the study of the functional role of the appendix in maintaining gut microbiota homeostasis and pathogenesis of gut colitis and malignancies, as well as for the potential development of drug targeting therapies.

The presented protocol describes a facile surgical removal of the appendix (caecal patch) in a mouse followed by the induction of inflammatory bowel disease-associated colorectal cancer. This murine appendectomy model enables investigation of the biological role of the appendix in the pathogenesis of human gastrointestinal disease.

慢性结肠炎相关结肠直肠癌的肿瘤切除术模型，通过卡卡贴片的精确定位

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal ...

Establishing In Situ Closed Circuit Perfusion of Lower Abdominal Organs and Hind Limbs in Mice

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small size of mouse organs, ex vivo perfusion of bone, bladder, skin, prostate, and reproductive organs is challenging or not feasible. Here, we report for the first time an in situ lower body perfusion circuit in mice that includes the above tissues, but bypasses the main clearance organs (kidney, liver, and spleen). The circuit is established by cannulating the abdominal aorta and inferior vena cava above the iliac artery and vein and cauterizing peripheral blood vessels. Perfusion is performed via a peristaltic pump with perfusate flow maintained for up to 2 h. In situ staining with fluorescent lectin and Hoechst solution confirmed that the microvasculature was successfully perfused. This mouse model can be a very useful tool for studying pathological processes as well as mechanisms of drug delivery, migration/metastasis of circulating tumor cells into/from the tumor, and interactions of immune system with perfused organs and tissues.

A protocol is described for in situ perfusion of the mouse lower body, including the bladder, the prostate, sex organs, bone, muscle and foot skin.

在小鼠中建立下腹部器官和后肢的原点闭路灌注

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small ...