我们感谢谁对这个模型工作的许多学生和研究人员，以提高其有效性，并了解其作用机制。特别感谢博士的。玛丽·约翰逊（马里兰大学巴尔的摩分校），Nitza登堡 - 科恩（施奈德曼儿童医院，佩塔提克瓦-Tikva的，以色列），张朝阳（爱因斯坦医学院，布朗克斯，NY），和Valerie Touitou（总医院Salpetrie，法国巴黎）。这项研究是由RO1 EY015304为SLB部分资助。

该协议被批准马里兰机构动物护理和利用委员会的大学（IACUC，巴尔的摩，MD，USA） 1.实验装置<ol><li>使由透明光学级圆形直径7毫米的有机玻璃，厚3mm的定制设计的隐形眼镜。切用钻床的圆形透镜。使用标准钻头以使内曲面，最后抛光使用超微粒度（3000分之1000）的接触透镜抛光外侧和内侧曲线。 </li><li>预先在pH 7.4的磷酸盐缓冲盐水（PBS）制备的2.5mM玫瑰红（RB），过滤消毒用0.45微米注射器式滤器并储存在-20℃1ml等份在一个不透光的容器中长达6个月。 注意:使用白化远系繁殖的雄性动物，如只SD的最小的应变依赖性反应的差异，使增加的易于诱导，并减少可与发情个循环发生的变异NG女性。 </li><li>设置了倍频钕钇铝石榴石（FD-YAG）眼科医疗激光，其产生532nm的激光。安装使用标准的眼科激光适配器上海牙，斯特赖特眼科裂隙灯激光。这市售设备时使动物的眼睛和激光光斑应用程序同时可视化。激光医疗也有适当的对焦和中心定位使用相同的光斑尺寸激光感应瞄准光束。激光功率参数如下: <ol><li>对于大鼠rNAION感应:使用500微米的光斑尺寸/ 50毫瓦的激光功率/ 1000毫秒时间/ 1000毫秒间隔。 </li><li>对于鼠标rNAION感应:改变光斑大小为300微米的较小的视盘和离开其他参数相同的鼠设置。 </li><li>经常使用的激光功率计，以保证激光功率输出。 </li></ol></li></ol> 2.实验步骤<ol><li>打开激光功率并设置适当的激光参数。暖激光使用前至少五分钟。 </li><li>称重动物，以确定氯胺酮/赛拉嗪和RB染料的适当剂量。由80毫克/毫升氯胺酮和4mg / ml的甲苯噻嗪1毫升/千克混合物的腹膜内注射麻醉动物。 </li><li>离开动物在笼子里加热，直至完全麻醉。检查到厌恶刺激（尾或脚趾捏）没有反应。检查动物麻醉深度每10分钟。 </li><li>扩张动物的瞳孔以1％托并用0.5％丙对卡因麻醉眼的表面上。如果使用着色的动物，如龙埃文斯，2.5％neosynephrine滴眼液将增加瞳孔扩大。 </li><li>用剪刀剪下靠近枪口上侧面的胡须被诱导，以避免遮挡住了。 </li><li>把1％甲基纤维素或其他眼科耦合下降到定制的隐形眼镜内的一滴，然后应用到镜头Ť他眼睛鼠。 </li><li>放置动物调至裂隙灯的高度的平台上。使眼球垂直于裂隙灯和激光束以45°角设置动物的头。 </li><li>通过眼科裂隙灯形象化的眼球。确保瞄准光束的大小是否合适，以及重点突出，直接集中在可视化视神经。速度方面地安装在裂隙灯眼睛件与定制适配器之一 - 使用数字照相机具有高的ASA（2,000 1,200）拍摄视网膜眼底。 注意:要看到脉络膜血管的能力，揭示了视网膜的透明度。这是为了能够检测后视网膜缺血，如果它发生可混淆的解释的重要标志。 rNAION是视神经缺血，这导致分离的RGC损失，而在视网膜损伤视网膜缺血的结果影响到所有视网膜内层的细胞。 </li><li>任选地，进一步的图像在视网膜和视神经使用谱域光学相干断层扫描（SD-10月8日），以评估未引起注意。扫描连接面 （ 图3B），以及通过在视网膜（ 图3C）7剖扫描。对SD-OCT成像使用用于激光感应同一隐形眼镜。 </li><li>注射1毫升/公斤静脉注射RB尾静脉，并等待30秒，然后激活激光功率。这次延迟使得RB在整个循环均匀分布。我们用一个50毫瓦的激光脉冲在同一秒/脉冲。更大的能量（≥60 MW）会损害视网膜或引起视网膜血管缺血。 注意:确保每个人都有一双合适的阻塞波长的激光安全过滤眼镜，以防止杂散激光进入研究者的眼球。 注意:激光给药必须迅速静脉注射RB注射后给予因为染料被从循环迅速消灭。较长的激光诱导，在更严重的视神经缺血。一般情况下，动物被赋予7 - 12秒脉冲在快速连续。激光的与循环染料的接触使视神经血管美丽的金色的光晕，可以通过裂隙灯（ 图4B）可见。这证明了染料全身注射和分布到血流中。如果发光微弱，或根本（ 图4A）无，染料不静脉注射。在这种情况下，不立即给予第二次注射，因为动物将需要用于再注入之前至少两天恢复。 </li><li>感应后立即移除隐形眼镜。覆盖眼科三联抗生素（抗生素软膏/多粘菌素/杆菌肽）软膏地塞米松双眼，然后将动物在严密观察下一个安置笼37℃加热垫，直到完全康复。 <ol><li>清洁隐形眼镜用蒸馏水W¯¯亚特并擦干用非研磨性清洁布以备将来使用。 </li><li>诱导两天后评估由双方眼底彩色照相和SD-OCT分析8视神经水肿。 注:视神经水肿程度的比较给出了视神经缺血的严重程度的估计。在两天内，视神经盘缘模糊，以及视网膜静脉稍微扩张，与对侧（未诱导）眼相比较。两个，其余四周进行电图（ERG）和闪光视觉诱发电位（VEP 11）发布诱导电分析。 </li></ol></li></ol>

<ol>
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作者什么都没有透露。

虽然有若干的视神经损伤（视神经损伤12，视神经横断13和PION 14）的机型，rNAION模型是人性化，适应大鼠和小鼠。它更类似于NAION的人体临床状况。这种情况包括渐进式前视神经水肿，前路视神经骨筋膜室综合征，局灶性脑缺血轴突的，孤立的视网膜神经节细胞轴突的破坏和损失在延长的时间过程。目前报告给出了rNAION感应适当的步骤，诱导期间讨论的潜在问题，并且描述...

非动脉炎性前部缺血性视神经病（NAION）是视神经（ON）1的前部的局灶性缺血病变。 NAION是在50 2岁以上的个体突然视神经相关的视力丧失的最常见的原因。机理被认为是，其导致神经内水肿一个室综合征，并且使得毛细管视神经内供给轴突压缩3。 因为在ON实际上是一种中枢神经系统（CNS）道，啮齿动物NAION（rNAION）模型可以用于机制和响应研究分离的CNS白质笔。因此，rNAION模型可能在解剖与白质中风相关的损害相关的许多问题非常有用。它可以用来评估在白质行程不同神经保护策略和代理。 之一的模型的最吸引人的特点是，它是无痛，无创的过程。激光功率可被调节，以产生不同程度的局部缺血损伤的。另一个特点是，它依赖于激光诱导超氧自由基损伤毛细血管内皮，产生累进毛细管功能障碍。它是被认为是非常相似的，导致NAION机制此功能障碍和渐进水肿。研究已经表明，它不会导致直接毛细管凝结，但通过至少两种机制工作的:过氧化物诱导的死亡和一些毛细管的剥离内皮细胞4，和激活的B细胞的的NFkB（核因子kappa轻链增强剂）相关的炎症性上调在剩余的内皮，具有增加的跨细胞膜流体输送入间质5。视神经毛细血管受压所致间液积聚导致视神经缺血头的关闭。示意图图片所示图2，rNAION模型可在大鼠和小鼠物种6,7-都可以使用，并可以在其严重性的程度而改变，从轻微损伤视神经的完整，但无痛破坏和视网膜等视网膜中央动脉阻塞（CRAO）。

<table border="0" cellpadding="0" cellspacing="0" width="600">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50mW 532nm laser</td>
			<td style="width:99px;">Iridex</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Standard Ophthalmic Laser</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">0-100mW 532nm laser</td>
			<td style="width:99px;">Laserglow technologies</td>
			<td style="width:119px;"></td>
			<td>Substitute for iridex</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laser slit lamp adapter</td>
			<td style="width:99px;">Iridex</td>
			<td style="width:119px;"></td>
			<td>SMA coupled adapter for laser output</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cpherent Fieldmate laser meter with thermopile sensor</td>
			<td style="width:99px;">Coherent</td>
			<td style="width:119px;"></td>
			<td>others also appropriate</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Ophthalmic Examing Slit lamp biomicroscope</td>
			<td style="width:99px;">VArious</td>
			<td style="width:119px;"></td>
			<td>Haag-Streit is the best; cheaper versions available on ebay</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Rose Bengal</td>
			<td style="width:99px;">Sigma</td>
			<td style="width:119px;">330000-1G</td>
			<td>Photoinducing agent</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Fundus Contact lens or glass cover slip</td>
			<td style="width:99px;">custom/Cantor and Nissel (UK)</td>
			<td style="width:119px;"></td>
			<td>Custom designed planoconvex plastic lens for eye exam and induction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tropicamide 1%</td>
			<td style="width:99px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tamiya polishing compound</td>
			<td style="width:99px;">Tamiya, INC</td>
			<td align="right" style="width:119px;">87068</td>
			<td>polishing contact lens</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">2.5% Hypromellose (Goniovisc)/1% Methycellulose</td>
			<td style="width:99px;">HUB Pharmaceuticals</td>
			<td style="width:119px;"></td>
			<td>contact lens coupling agent</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">2.5% Neosynephrine Ophthalmic drops</td>
			<td style="width:99px;">Alcon labs</td>
			<td style="width:119px;"></td>
			<td>pupil dilating agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tropicamide 1%</td>
			<td style="width:99px;">Alcon labs</td>
			<td style="width:119px;"></td>
			<td>pupil dilating agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">0.5% Proparacaine</td>
			<td style="width:99px;">Alcon labs</td>
			<td style="width:119px;"></td>
			<td>topical Anesthetic</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">30ga fused needle insulin syringe</td>
			<td style="width:99px;">Various</td>
			<td style="width:119px;">Various</td>
			<td>for intravenous injection of rose bengal</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Ophthamic Antibiotic ointment with dexamethasone added (Triple antibiotic ointment)</td>
			<td style="width:99px;">Various</td>
			<td style="width:119px;">Various</td>
			<td>Apply after induciton to minimize corneal scarring</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Heidelberg Corporation Spectral domain-Optical Coherence Tomograph</td>
			<td style="width:99px;">Heidelberg Corportion</td>
			<td style="width:119px;"></td>
			<td>For Optical coherence measurements baseline and post-induction; not essential for induction</td>
		</tr>
	</tbody>
</table>

the rodent model of nonarteritic anterior ischemic optic neuropathy (rnaion)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

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The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

非动脉炎性前部缺血性视神经病变的啮齿动物模型（rNAION）

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

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Research

JoVE Journal

Medicine

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

8.8K 次观看.  University of Maryland-Baltimore School of Medicine. The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

视频: The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

缺血性心力衰竭在临床前模型的转基因

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

科研

医学

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

光化学诱导后部缺血性视神经病变的大鼠模型

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

在大鼠缺血性皮肤创伤模型的演示

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

An Intact Pericardium Ischemic Rodent Model

This protocol has shown that the pericardium and its contents play an essential anti-fibrotic role in the ischemic rodent model (coronary ligation to induce myocardial injury). The majority of pre-clinical myocardial infarction models require the disruption of pericardial integrity with loss of the homeostatic cellular milieu. However, recently a methodology has been developed by us to induce myocardial infarction, which minimizes pericardial damage and retains the heart&#39;s resident immune cell population. An improved cardiac functional recovery in mice with an intact pericardial space following coronary ligation has been observed. This method provides an opportunity to study inflammatory responses in the pericardial space following myocardial infarction. Further development of the labeling techniques can be combined with this model to understand the fate and function of pericardial immune cells in regulating the inflammatory mechanisms that drive remodeling in the heart, including fibrosis.

This protocol outlines the steps for inducing myocardial infarction in mice while preserving the pericardium and its contents.

完整的心包缺血性啮齿动物模型

This protocol has shown that the pericardium and its contents play an essential anti-fibrotic role in the ischemic rodent model (coronary ligation to ...

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a considerable proportion of coronary artery disease patients remain symptomatic. Gene therapy-mediated therapeutic angiogenesis offers a novel therapeutic method for improving myocardial perfusion and relieving symptoms. Gene therapy with different angiogenic factors has been studied in few clinical trials. Due to the novelty of the method, the progress of myocardial gene therapy is a continuous path from bench to bedside. Therefore, large animal models are needed for evaluating the safety and efficacy. The more the large animal model identifies the original disease and the endpoints used in clinics, the more predictable outcomes are from clinical trials. Here, we introduce a large animal model for evaluating the efficacy of the gene therapy in the ischemic porcine heart. We use clinically relevant imaging methods such as ultrasound imaging and 15H2O-PET. For targeting the gene transfers into the desired area, electroanatomical mapping is used. The aim of this method is: (1) to mimic chronic coronary artery disease, (2) to induce therapeutic angiogenesis at hypoxic areas of the heart, and (3) to evaluate the safety and efficacy of the gene therapy by using relevant endpoints.

Myocardial gene therapy for ischemic heart disease holds great promise for future therapeutics. Here, we introduce a large animal model for evaluating the efficacy of gene therapy in the ischemic heart.