本研究由中国国家自然科学基金U21A20368（L.Y.）、82101000（H.W.）和82100982（F.L.）以及四川省科技计划2023NSFSC1499（H.W.）资助。所有原始数据和图像均包含在本文中。

本研究的实验程序经四川大学华西口腔医学院伦理委员会（WCHSIRB-D-2021-125）批准。成年C57BL / 6小鼠购自中国成都的Gempharmatech实验动物公司。上颌第一磨牙的整个牙冠在出生后 21 天萌出。手术小鼠的年龄应大于21天，具有正常活力11。在这里，使用 6 至 8 周龄的小鼠进行建模。 图 1 是一个流程图，显示了所使用的协议。
1.术前准备（图2）
<ol><li>获取以下仪器:立体显微镜、固定板、医用胶带、口腔堵嘴、直径0.6mm的微创牙科毛刺、牙科高速牙科手机、8# C+锉、加热垫、1 mL注射器、无菌棉球、眼钳。</li>
	<li>获得以下药物:麻醉混合物，兽药膏。</li>
</ol>2.口塞的准备
<ol><li>通过腹膜内注射麻醉混合溶液（10%盐酸氯胺酮+ 5%甲苯噻嗪+ 85%无菌等渗盐水）以0.007mL / g体重称重并麻醉小鼠，并通过脚趾捏法确认适当的麻醉。将眼科润滑膏涂抹在眼睛上，以防止手术时因干燥而造成眼睛受伤。 注意:医用帽子、口罩、手套和工作服以及其他基本保护措施是必要的。确保手术环境和小鼠室清洁安全。在整个过程中，用于热支撑的加热垫是必要的。</li>
	<li>如下所述准备嘴巴堵嘴（图3）。<ol><li>获得以下材料:直径为 8 μm 的正畸弓丝、年轻的环弯曲钳、重型钢丝钳、记号笔、长度为 3 mm、横截面直径为 1 mm 的橡胶帽。</li>
		<li>首先，用左手拉直弓形线进行固定，右手的拇指、食指稍微弯曲以抵挡线的弧度。重复此动作几次，将有助于弯曲到正确的三维角度。</li>
		<li>使用Yong环弯曲钳将梯形（图3C，a-l-k-b）的顶部边缘（图3G，a-i）弯曲在弓形丝的中点长约8 mm。确保点 a（图 3）位于钳嘴的边缘。</li>
		<li>用左手握住钳子，用右手拇指和食指夹住弓线的自由端，从a点弯曲弓线，形成约120°的角度。在点i处复制上一个操作（图3G）。将弓形线放在水平台上而不撬动，检查弓形线是否在一个平面上。</li>
		<li>每侧留出约 9 毫米的长度（图 3D、a-b、l-k），并使用与步骤 2.2.4 相同的技能将自由端弯曲到 75° 角，同时确保每条边都在一个平面上。用钳嘴的尖端弯曲这个锐角。</li>
		<li>找到距离 b 点约 5 毫米的 c 点。按照相同的技巧在点 c 上弯曲 105° 角。在距离 c 点 5 毫米的 d 点再弯曲 105° 角。距 d 点约 4.5 mm，找到 e 点。在点 e 处弯曲自由端以形成约 100 -105° 的角度（图 3E）。 注意:我们使用的 6-8 周龄的 C57 小鼠约为 20 g。5毫米的间距不仅可以卡住小鼠的上下颚而不动弹，而且不会压迫小鼠的皮肤而引起不适。如果使用其他种类或年龄的小鼠，请根据实际情况调整c-d和i-h部分的长度（图3E，G）。</li>
		<li>为下颌部分弯曲一个额外的压舌板（图3G，j-i-h-g）。</li>
		<li>a-b-c零件在l-k-j零件上的重复弯曲步骤。同时夹紧 i-k 和 k-j 零件，并在 j 点弯曲自由端，使其垂直于 i-k-j 平面。夹紧点 i 距点 j 5 mm，弯曲自由端以使其平行于 i-k-j 平面和 c-d 部分（图 3H）。</li>
		<li>从 i 点起留 5 mm 长度，在 h 点处，将拱形弯曲垂直于 i-h 部分并平行于 j-i-h 平面。在距点 h 5 毫米处找到点 g。夹紧 j-i-h-g 平面并弯曲对称于 k-j 部分的自由端。然后，点f之后的自由端应与点e的自由端对称（图3H）。</li>
		<li>将橡胶帽放在自由端（图3F）。</li>
	</ol></li>
</ol>3. 固定
<ol><li>将小鼠仰卧固定在固定板上，四肢用皮胶带固定。用拇指和食指压缩嘴巴的自由端。</li>
	<li>将鼠标前门牙固定在两条手臂的梯形凹槽中。确保带有压舌板的手臂用于下颌骨。调整嘴巴堵嘴，确保老鼠的舌头被固定但不会缺血。</li>
</ol>4. 牙齿评估
<ol><li>确保手术时上颌第一磨牙应没有龋齿、外伤和牙龈生成。确保周围的牙龈没有发红、肿胀或瘘管。确保对面的牙齿健康，并可以作为健康的对照组。</li>
</ol>5. 纸浆暴露
<ol><li>使用牙齿毛刺以 20,000 rpm 的速度在上颌第一磨牙的咬合侧钻孔。确保去除牙釉质。仅在牙本质的浅层中保持牙科手机的操作，以防止对牙髓组织进行过度热刺激12.</li>
	<li>同时，在手术过程中每 3 分钟使用注射器将生理盐水滴在牙齿上，以防止过热。</li>
	<li>将 8# 或 10# C+ 锉刀放在钻坑的最低位置，然后刺穿最后一个牙本质，露出牙髓室。当局部牙本质被彻底去除时，会有一种明显的坠落感。不要探得太深，否则牙髓组织可能会从牙髓室中带出。</li>
	<li>清洁牙齿周围的碎片。取下嘴巴堵嘴;手术已经完成。使用对面的上颌第一磨牙作为对照，无需手术。</li>
</ol>6. 术后护理
<ol><li>手术后，皮下给予卡洛芬（5mg / kg），并将小鼠置于化学热加热垫上，以俯卧姿势，直到从麻醉中恢复。喂老鼠并提供饮用水。恢复过程应该受到监督。在小鼠完全恢复之前，其他动物不应在同一腔室中。</li>
</ol>7. 样本采集和储存
<ol><li>在手术后 24 小时或根据实验9 的任何其他时间点，在深度麻醉条件下对颈椎脱位的小鼠实施安乐死。用眼科剪刀剪开附着在上颌骨和颧骨上的骨骼肌。去除骨骼、额骨和软组织，取出带有上颌磨牙的蚋臼片。 注意:根据 He 等人的说法，建议在手术后不到 72 小时收集牙髓炎样本，以避免牙髓组织广泛坏死13。</li>
	<li>矢状地将蚊蚋石分成两半，并将组织储存在PBS（pH 7.4）中的4%多聚甲醛中，在4°C下进行24小时固定。</li>
</ol>8. 组织学分析
<ol><li>用磷酸盐缓冲盐水（PBS）洗涤组织，并在每日更换的5%EDTA和4%蔗糖的PBS（pH 7.4）脱钙溶液中脱钙，在4°C下2-4周10。</li>
	<li>将 1/2 蚊蚋石嵌入石蜡中，并确保没有牙齿的矢状面位于组织盒的底部。</li>
	<li>用石蜡切片机将石蜡块切成5μm厚的切片。根据显微镜下观察到的近端、远端、上部和下部关系调整石蜡块的角度，以确保可以切割完整的冠髓和第一磨牙的穿孔。</li>
</ol>

使用新型 Mouth-Gag 创建牙髓炎症小鼠模型 

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作者声明没有利益冲突。

作为牙齿内孤立的软组织，牙髓在维持牙齿的生物活性方面发挥着至关重要的作用，但仍然高度敏感。保留这种重要的牙髓已成为最近牙髓治疗中首选的初始方法，因此需要全面了解牙髓的炎症机制16。炎症微环境的时空波动和牙髓炎中驻留细胞类型之间的相互作用使其通过 体外 研究的研究变得复杂11.相反， 体内 研究通过复制在人类中发现的生?...

牙髓是牙齿的一个组成部分，负责多种基本功能，如营养供应、牙本质形成、感觉功能和防御反应1。然而，被硬组织包围的牙髓容易受到深龋、牙髓炎、创伤或后续治疗的伤害和损伤 2,3。功能性牙髓的缺失会增加牙齿脆弱的风险4.此外，年轻恒牙牙髓活力的丧失会对牙齿成熟产生不利影响，而目前的假牙技术无法恢复健康牙髓提供的神经反馈4.这种情况促使研究人员探索管理发炎牙髓的替代解决方案，而不仅仅是去除。
2007 年，Murray 等人开始将组织工程应用于再生牙髓学，从而引发了人们对牙髓保存和再生的兴趣5。然而，发炎的牙髓组织带来了挑战，因为细胞会释放 IL-6 等炎症因子，IL-6 会招募炎症细胞并导致细胞坏死、牙髓活力丧失和功能恢复并发症 6,7。因此，了解炎症和相关的细胞死亡对于保护重要牙髓的进展至关重要。已经进行了许多实验来探索炎症牙髓在体内或体外的分子生物学 8,9。尽管像 2D 或 3D 细胞培养这样的体外实验已经发展了多年，并且已经变得成熟并广泛用于测试牙髓细胞对炎症因子的反应，但这些实验无法反映牙髓组织与全身免疫系统之间的相互作用10。如果正在研究的现象来源于其他组织来源的细胞，如免疫、血管和神经系统，那么纯牙髓细胞培养将导致死胡同。因此，体内实验是非常必要和具有参考意义的。
小鼠因其成本效益、高生育率和活力而日益成为 体内炎症研究中的常见选择。然而，目前尚无针对小鼠牙髓炎模型的综合方案，可作为参考。小鼠的小尺寸和它们对刺激的敏感性在实验过程中构成了重大挑战。观察隐藏在小鼠口腔深处的微小牙齿通常需要使用悬臂显微镜，尽管桌面显微镜在实验室中更为普遍。没有张口器需要其他人的帮助。为了解决这个问题，该小组使用现成的材料设计了一种口腔堵嘴器，旨在为构建小鼠牙髓炎模型提供标准化和可重复的协议。本文详细介绍了该过程，涵盖 C57 小鼠的术前准备、固定、牙髓暴露手术和样本采集。该协议建议使用口腔堵嘴，提供有关其结构、生产和应用的信息，以方便其他研究人员复制该程序。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Animal</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C57/B6J mice</td>
			<td>Gempharmatech Experimental Animals Company</td>
			<td>C57/B6J</td>
			<td>For the establishment of pulp exposure</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>Chengdu Xinjin Shifeng Medical Apparatus &#38; Instruments Co. LTD.</td>
			<td>SB1-074(IV)</td>
			<td>Apply in drug injection.</td>
		</tr>
		<tr>
			<td>8# C+ file</td>
			<td>Readysteel</td>
			<td>0010047</td>
			<td>Apply in exposing the roof of pulp chamber.</td>
		</tr>
		<tr>
			<td>Anesthesia Mix solution</td>
			<td></td>
			<td></td>
			<td>10% ketamine hydrochloride+ 5% xylazine + 85% sterile isotonic saline.&#160;</td>
		</tr>
		<tr>
			<td>DAPI Staining Solution</td>
			<td>Beyotime</td>
			<td>C1005</td>
			<td>Apply in immunofluorescence staining for counter-staining of nucleus.</td>
		</tr>
		<tr>
			<td>Dental high-speed dental handpiece</td>
			<td>Jing yuan electronic commerce technology</td>
			<td>WJ-422</td>
			<td>Apply in pulp exposure.</td>
		</tr>
		<tr>
			<td>Heavy wire cutter</td>
			<td>Jirui Medical Instrument Co., Ltd.</td>
			<td>none</td>
			<td>Apply inarc cutting.</td>
		</tr>
		<tr>
			<td>Hematoxylin and Eosin Stain kit</td>
			<td>Biosharp</td>
			<td>BL700B</td>
			<td>For the histological analysis of the slides.</td>
		</tr>
		<tr>
			<td>IL-6 antibody</td>
			<td>Novus</td>
			<td>NBP2-89149</td>
			<td>Apply in immunofluorescence staining to detect the inflammation of the dental pulp.</td>
		</tr>
		<tr>
			<td>Ketamine(Ketamine hydrochloride)</td>
			<td>Vet One, Boise, Idaho, USA</td>
			<td>C3N VT1</td>
			<td>100mg/kg, IP. Apply in nesthetization.</td>
		</tr>
		<tr>
			<td>Medical tap</td>
			<td>3M</td>
			<td>1530</td>
			<td>Apply in mice immobilization.</td>
		</tr>
		<tr>
			<td>Orthodontic arch wire&#160;</td>
			<td>Shanghai Wei Rong Medical Apparatus Co. LTD.</td>
			<td>K417</td>
			<td>Diameter of 8&#181;m</td>
		</tr>
		<tr>
			<td>Round dental burr (0.6 mm)</td>
			<td>Shofu global</td>
			<td>072208</td>
			<td>Apply in removing enamel and shallow layer of dentin.</td>
		</tr>
		<tr>
			<td>Young loop bending plier</td>
			<td>Jirui Medical Instrument Co., Ltd.</td>
			<td>none</td>
			<td>Apply in arc bending.</td>
		</tr>
	</tbody>
</table>

establishment of a murine pulp exposure model with a novel mouth-gag for pulpitis research

牙髓炎是自然牙齿脱落的常见原因，会导致发炎牙髓中的坏死和生物活性丧失。揭示牙髓炎的机制及其有效治疗是牙髓病学研究的持续重点。因此，了解牙髓内的炎症过程对于改善牙髓保存至关重要。与其他体 外 实验相比，鼠牙髓炎模型为观察牙髓炎的病理进展提供了更真实和遗传多样化的背景。然而，尽管小鼠具有成本效益和可及性，但由于它们的体积小、协调性差和耐受性低，使用小鼠会带来困难，使口腔内和牙科手术复杂化。该协议引入了一种新颖的设计和应用，即口腔堵嘴以暴露小鼠牙髓，从而促进更有效的口内手术。口腔堵嘴由大多数牙医都可以轻松使用的牙弓组成，可以显着加快手术准备，即使是第一次手术也是如此。Micro-CT、苏木精-伊红（hematoxylin-eosin，HE）染色和免疫荧光染色（immunofluescing sing）检测形态和细胞表达的变化。本文的目的是帮助研究人员建立一种更具可重复性且要求更低的程序，以使用这种新型的口腔堵嘴创建牙髓炎症模型。

在3,6-8周龄C57BL / 6小鼠的右上颌第一磨牙上进行上述程序，同时保留左上颌第一磨牙作为对照。空白对照、12 h牙髓炎和24 h牙髓炎样品的组织学和免疫荧光结果用于示范。
根据 Goldman 等人 15 的 CT 分析方案，通过 图 4 A-C 中的显微 CT 和重建建模确认了牙髓暴露。来自对照侧和手术侧的上颌第一磨牙的矢状面切片都进行了 HE ?...

Watch this Scientific Journal Video about Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research at JoVE.com

作者聚焦:通过改进的小鼠模型增强牙髓研究

本文提出了一种简化的方案，用于使用创新的口堵在小鼠中建立牙髓炎模型，然后进行随后的组织学分析。

基于新型口堵的鼠牙髓暴露模型的建立及其在牙髓炎研究中的应用

Pulpitis, a common cause of natural tooth loss, leads to necrosis and loss of bioactivity in the inflamed dental pulp. Unraveling the mechanisms ...

2D或3D细胞培养等单个实验已被广泛用于测试牙髓细胞的反应。这些实验不能反映牙髓组织与全身免疫系统之间的相互作用。因此，体内实验在牙髓研究中越来越受欢迎。C57小鼠因其成本效益、高生育率和活力而日益成为牙髓炎症研究中的常见选择。然而，小鼠体积小和缺乏协调性在实验过程中带来了重大挑战。固定过程是提高技术门槛的重要因素。我们使用现成的材料和简单的组装设计了一种新颖的口腔堵嘴器，以帮助研究人员进行牙髓暴露手术。根据我们的研究，这种口腔堵嘴可以大大缩短初学者固定嘴巴和暴露小鼠上颌第一磨牙所需的时间。

establishment-murine-pulp-exposure-model-with-novel-mouth-gag-for

Research

JoVE Journal

Medicine

Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research

859 次观看.  Sichuan University. 2D或3D细胞培养等单个实验已被广泛用于测试牙髓细胞的反应。这些实验不能反映牙髓组织与全身免疫系统之间的相互作用。因此，体内实验在牙髓研究中越来越受欢迎。C57小鼠因其成本效益、高生育率和活力而日益成为牙髓炎症研究中的常见选择。然而，小鼠体积小和缺乏协调性在实验过程中带来了重大挑战。固定过程是提高技术门槛的重要因素。我们使用现成?...

视频: 作者聚焦:通过改进的小鼠模型增强牙髓研究

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

科研

医学

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

轻度脑外伤的幼年大鼠一种新模式

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

的不可逆和可逆单侧输尿管梗阻小鼠模型

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Establishment and Characterization of UTI and CAUTI in a Mouse Model

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. Females are disproportionately afflicted by UTI: 50% of all women will have a UTI in their lifetime. Additionally, 20-40% of these women who have an initial UTI will suffer a recurrence with some suffering frequent recurrences with serious deterioration in the quality of life, pain and discomfort, disruption of daily activities, increased healthcare costs, and few treatment options other than long-term antibiotic prophylaxis. Uropathogenic Escherichia coli (UPEC) is the primary causative agent of community acquired UTI. Catheter-associated UTI (CAUTI) is the most common hospital acquired infection accounting for a million occurrences in the US annually and dramatic healthcare costs. While UPEC is also the primary cause of CAUTI, other causative agents are of increased significance including Enterococcus faecalis. Here we utilize two well-established mouse models that recapitulate many of the clinical characteristics of these human diseases. For UTI, a C3H/HeN model recapitulates many of the features of UPEC virulence observed in humans including host responses, IBC formation and filamentation. For CAUTI, a model using C57BL/6 mice, which retain catheter bladder implants, has been shown to be susceptible to E. faecalis bladder infection. These representative models are being used to gain striking new insights into the pathogenesis of UTI disease, which is leading to the development of novel therapeutics and management or prevention strategies.

The ability to model urinary tract infections (UTI) is crucial in order to be able to understand bacterial pathogenesis and spawn the development of novel therapeutics. This work&#8217;s goal is to demonstrate mouse models of experimental UTI and catheter associated UTI that recapitulate and predict findings seen in humans.

建立和UTI和CAUTI表征在小鼠模型

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

Working with Human Tissues for Translational Cancer Research

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human specimens, precious samples must be handled with the greatest care in order to ensure integrity of organs, tissues, and cells to the highest degree. Unfortunately, tissue processing by definition requires extraction of tissues from the host, a change which can cause great cellular stress and have major repercussions on subsequent analyses. These stresses could result in the specimen being no longer representative of the site from which it was retrieved. Therefore, a strict protocol must be adhered to while processing these specimens to ensure representativeness. The desired assay(s) must also be taken into consideration in order to ensure that an optimal technique is used for sample processing. Outlined here is a protocol for tissue retrieval, processing and various analyses which may be performed on processed tissue in order to maximize downstream production from limited tissue samples.

Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples.

与人体组织转化为癌症研究工作

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

动静脉内瘘失败的新型小鼠模型:在详细介绍手术过程

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening

Wear particle-induced osteolysis is a major cause of aseptic loosening in arthroplasty failure, but the underlying mechanism remains unclear. Due to long follow-ups necessary for detection and sporadic occurrence, it is challenging to assess the pathogenesis ofparticle-induced osteolysis in clinical cases. Hence, optimal animal models are required for further studies. The murine model of calvarial osteolysis established by exposure to CoCrMo particles is an effective and valid tool for assessing the interactions between particles and various cells in aseptic loosening. In this model, CoCrMo particles were first obtained by high-vacuum three-electrode direct current and resuspended in phosphate-buffered saline at a concentration of 50 mg/mL. Then, 50 &#181;L of the resulting suspension was applied to the middle of the murine calvaria after separation of the cranial periosteum by sharp dissection. After two weeks, the mice were sacrificed, and calvaria specimens were harvested; qualitative and quantitative evaluations were performed by hematoxylin and eosin staining and micro computed tomography. The strengths of this model include procedure simplicity, quantitative evaluation of bone loss, rapidity of osteolysis development, potential use transgenic or knockout models, and a relatively low cost. However, this model cannot to be used to assess the mechanical force and chronic effects of particles in aseptic loosening. Murine calvarial osteolysis model generated by exposure to CoCrMo particles is an ideal tool for assessing the interactions between wear particles and various cells, e.g., macrophages, fibroblasts, osteoblasts and osteoclasts, in aseptic loosening.

This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening.

CoCrMo 颗粒暴露的小鼠颅骨骨溶解模型的构建与评价

Wear particle-induced osteolysis is a major cause of aseptic loosening in arthroplasty failure, but the underlying mechanism remains unclear. Due to long ...

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 ...

Establishing a Silicosis Rat Model via Exposure of Whole-Body to Respirable Silica

The major cause of silicosis is the inhalation of silica in the occupational environment. Despite some anatomical and physiological differences, rodent models continue to be an essential tool for studying human silicosis. For silicosis, the classic pathological process needs to be inducible via the inhalation of freshly generated quartz particles, which means specifically inducing human occupational disease. This study described a technique to establish and effectively mimic the pathological dynamic evolution process of silicosis. Further, the technique had good repeatability with no surgery involved. The inhalation exposure system was fabricated, validated, and used for toxicology studies on respirable particle inhalation. The critical components were as follows: (1) bulk dry SiO2 powder generator adjusted with an air-flow controller; (2) 0.3 m3 whole-body inhalation exposure chamber accommodating up to 3&#160;adult rats; (3) a monitoring and control system for regulating oxygen concentration, temperature, humidity, and pressure in real-time; and (4) a barrier and waste disposal system for protecting laboratory technicians and the environment. In summary, the present protocol reports the inhalation via the whole body, and the inhalation chamber created a reliable, reasonable, and repeatable rat silicotic model with low mortality, less injury, and more protection.

This study describes a technique to establish a silicosis rat model with the inhalation of silica through the whole body in an inhalation chamber. The rats with silicosis could closely mimic the pathological process of human silicosis in an easy, cost-effective manner with good repeatability.

通过全身暴露于可吸入二氧化硅来建立矽肺大鼠模型

The major cause of silicosis is the inhalation of silica in the occupational environment. Despite some anatomical and physiological differences, rodent ...