这项工作是由美国国立卫生研究院（R01DK58096到文森特Poitout）的支持。文森特Poitout持有加拿大研究主席在糖尿病和胰岛β细胞功能。贝德Zarrouki收到来自默克和礼来公司博士后奖学金。 Ghislaine FONTES是由加拿大糖尿病协会的博士后奖学金支持。

概述：该过程包括右颈静脉和左颈总动脉置管在全身麻醉下，允许7天的恢复期;导管连接使用一个旋转的泵和配重系统，使动物在笼子里自由移动;输注葡萄糖和/或脂 ​​肪乳剂（大豆油的乳液生成的混合物约80％unsaturated/20％的饱和脂肪酸时，注入肝素9）72小时。 1。右颈静脉和左颈总动脉Canulation <ol><li>消毒手术器械。 canulation管也必须冷灭菌使用液体消毒剂（2.6％戊二醛）前的程序。油管浸入一个蒸压容器16-24小时。冲洗，并用无菌蒸馏水彻底冲洗戊二醛使用前删除所有痕迹。 </li><li>称量大鼠，​​计算药物剂量： 卡洛芬5毫克/公斤：稀释1/10 =体重（克）所述0.001毫升SC（镇痛） 格隆0.01毫克/千克稀释1/10 = BW（七）所述0.0012毫升SC（抗胆碱） </li><li>麻醉大鼠用异氟醚和氧气。 </li><li>大鼠放置在其肚子上。剃须面积耳朵后面的肩膀基地。莱老鼠在它的后面。剃须区域脖子下的前爪。 </li><li>准备外科手术部位与洗必泰，酒精，和碘。管理药物。 </li><li>将大鼠的手术野。 </li><li>使用无菌技术，canulate右颈静脉和左侧颈总动脉，用PE-50 5 U的肝素生理盐水1 ml注射器填充有连接到插管。 50 U的肝素盐水冲洗canulas恢复期间避免凝血。使用减弱23G的针头。关闭每套管用23G针。 </li><li>手术后，削减约2.5毫米关闭底部门齿和大鼠在输液外套，以防止被吃掉canulas。 </li><li>提供氧气（1升/分钟，10分钟），以协助疏散异氟醚。 </li><li>将大鼠在加热笼，直到它完全清醒。 </li><li>操作只大鼠使用每一个输液条件（ 见表1）。 </li></ol> 2。手术后的护理（手术后的治疗和导管连接） <ol><li>称量大鼠第1天，手术后第2天。 </li><li>管理格隆（BW（克）×0.00048毫升）SC一次手术后第1天，第2天手术后两次。 </li><li>如果有必要，可以给予额外的支持治疗：液体，加热垫，湿饮食，氧疗，止痛药，抗胆碱能药物。 </li><li>手术后的第7天，大鼠重计算输液流速。 </li><li>将每只大鼠输液系统使用的系绳和旋转安装在一个笼子里格栅的顶部（图1）。</li><li> 5 U肝素盐水去除血块冲洗canulas。冲洗canulas，再一次与50 U的肝素盐水，以防止凝血。 </li><li>允许大鼠系绳适应和旋转输液开始前至少24小时。 </li></ol> 3。注入<ol><li>绘制0.15毫升的血，从每只大鼠的颈动脉并测量血糖。冲洗颈canulas。使用50 U的肝素盐水在每只大鼠的canulas都以防止凝血。 </li><li>血液样品转移到0.5 ml收集管中含有2％的EDTA。 10,000 rpm下离心2分钟，于-20℃下冻结等离子</li><li>填写两个60毫升的注射器各输液下面列出的条件。将注射器1靠前的位置在后卫位置上的泵泵和地方注射器2。 </li><li>一起加入每对解决方案与y轴的连接器和CO-EX T22管经过消毒。将注射器在哈佛33双注射器泵。 </li><li>更改笼底，并删除所有的食物从笼子格栅顶部。 </li><li>称取150克的标准饲料和地点上笼格栅顶部。 </li><li>将注射器旋转笼格栅。冲洗注射器，从线，以去除残留空气。 </li><li>之前所连接大鼠输液系统使用体重计算输液流速。 Microsoft Excel文件转换成毫升/小时的葡萄糖输注率（GIR）的价格计算。 </li><li>将泵设置为管理根据制造商的设定流速为60毫升注射器。输入的速度注射器1（前面注射器）和汇率注射器（注射器）。 </li><li>启动泵。 </li></ol> 4。监控<ol><li>启动泵后，确认有没有从旋转或从canulas的泄漏和不扭曲，输液管。也验证了，有没有气泡管子。 </li><li>经过3小时的输液，采取血液样本，监测血糖。重复后，6，24，48，和72小时的输液。作为预防措施，还监测血糖30，34，54和60小时后的输液。用一滴全血使用的是便携式的血糖监测仪，可以测量血糖。这限制了在输液过程中绘制的血液量，因此就避免了显着改变的血容量和/或血细胞比容。 </li><li>注射器1的汇率在被修改，根据血糖值维持血糖在220-250毫克/分升。 72小时输液过程中不会改变，因为游离脂肪酸维持在1 mmol / L的汇率注射器2 </li><li>输液条件是成对的，使收到的对照组动物的体积相当于为试验动物（ 表1）接收的体积。 </li><li>经过24小时的输液，改变笼底和权衡留在笼子里烧烤食物。返回吃剩的部分TØ笼格栅。 48小时后重复上述步骤。 </li><li>填充注射器每天需要在72小时输液。 </li><li>在输液过程中，始终遵循的大鼠炎症或不适的任何迹象。管理适当的照顾，如果需要的话。 </li></ol> 5。输液后安乐死<ol><li class="jove_content">输液72小时后，麻醉大鼠静脉注射0.5毫升（182毫克/公斤的氯胺酮和11.6毫克/公斤甲苯噻嗪1:2稀释，0.9％氯化钠）氯胺酮/甲苯​​噻嗪的鸡尾酒。 </li><li class="jove_content">脚趾捏反射是用来验证麻醉水平。额外的麻醉管理需要。 </li><li class="jove_content">当大鼠麻醉，打开腹腔手术剪。将大鼠放血，通过绘制从主动脉或腔静脉的注射器中的10-15毫升的血液。 </li></ol>

<ol>
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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minireview:+weapons+of+lean+body+mass+destruction:+the+role+of+ectopic+lipids+in+the+metabolic+syndrome.">Minireview: weapons of lean body mass destruction: the role of ectopic lipids in the metabolic syndrome.</a> Endocrinology. 144, 5159-5165 (2003).</li><li>Harmon, J. S., Gleason, C. E., Tanaka, Y., Poitout, V., Robertson, R. 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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+glucose+stimulates+early+response+gene+c-Myc+expression+in+rat+pancreatic+beta+cells.">High glucose stimulates early response gene c-Myc expression in rat pancreatic beta cells.</a> The Journal of Biological Chemistry. 276, 35375-35381 (2001).</li><li>Tang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose-induced+beta+cell+dysfunction+in+vivo+in+rats:+link+between+oxidative+stress+and+endoplasmic+reticulum+stress.">Glucose-induced beta cell dysfunction in vivo in rats: link between oxidative stress and endoplasmic reticulum stress.</a> Diabetologia. 55, 1366-1379 (2012).</li><li>Alonso, L. 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J., Froguel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+human+obesity.">The genetics of human obesity.</a> Nature Reviews. Genetics. 6, 221-234 (2005).</li><li>Fontes, G., Hagman, D. K., Latour, M. G., Semache, M., Poitout, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lack+of+preservation+of+insulin+gene+expression+by+a+glucagon-like+peptide+1+agonist+or+a+dipeptidyl+peptidase+4+inhibitor+in+an+in+vivo+model+of+glucolipotoxicity.">Lack of preservation of insulin gene expression by a glucagon-like peptide 1 agonist or a dipeptidyl peptidase 4 inhibitor in an in vivo model of glucolipotoxicity.</a> Diabetes Res. Clin. Pract. 87, 322-328 (2010).</li><li>Crawford, P. A., Schaffer, J. 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文森特Poitout是共同创始人和研究合约Bêtagenex公司，合同研究组织，提供输液模型在这篇文章中提出，作为一个商业服务。

虽然所采用的以往的研究慢性输注葡萄糖（ 例如 10-15）或脂质（ 如 16,17）在啮齿动物中，据我们所知，这两种燃料的联合输注已被发现在小鼠18。这里介绍的慢性输液模型提供了多种优势，在大鼠多种生物学功能的影响进行研究营养过剩。首先，它并不涉及遗传性肥胖的啮齿动物，因为人类常见的肥胖是19多基因遗传，因此，结果是更可能是在一般?...

向2型糖尿病的发病机制，通过改变一些器官的功能，在维持葡萄糖体内平衡，包括有牵连，长期升高的血糖和血脂水平的循环中已被提出，但不限于，胰腺β-细胞（评论在1）中。糖毒性假说认为慢性高血糖引起高血糖摆在首位，从而形成恶性循环，在2型糖尿病患者血糖控制的恶化加剧了β细胞的缺陷。同样，glucolipotoxicity的假说认为，由于经常在2型糖尿病患者中观察到，伴随升高的血糖和血脂水平的β细胞的不利。 破译长期升高的营养物质的有害影响胰岛β-细胞功能的细胞和分子机制的理解是关键2型糖尿病的发病机制中的1毫微克。为此，大量的研究已审查机制在孤立的胰岛体外克隆，胰岛素分泌细胞线，慢性营养过剩体外 。然而，这些模型系统的整个有机体中得到的结果的翻译是复杂的，特别是因为用于在培养的细胞或胰岛细胞的脂肪酸的浓度很少2 在体内的β细胞中的周边相匹配的循环水平。另一方面，营养过量的β细胞功能衰竭的机制已被被检查的糖尿病的啮齿动物模型中，例如由朱克糖尿病大鼠脂肪3,4，沙土鼠Psammomys大眼金枪鱼 5和高脂肪的饮食美联储鼠标6。然而，这些模型中，其特征在于由内在代谢异常，并且不能容易地适合于操纵血糖和/或脂质水平中的更多的控制和慢性的设置。为了能够改变血中营养水平的时间表在其他正常动物的天，我们已经开发出一种慢性输液正常大鼠模型，使我们能够检查血脂和血糖的影响，单独或组合，生理参数和函数7,8。

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>Saline 0.9%</td>
			<td>BD</td>
			<td>JB1324</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Dextrose 70%</td>
			<td>McKesson</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Intralipid 20%</td>
			<td>Fresenius Kabi</td>
			<td>JB6023</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Metricide (Glutaraldehyde 2.6%)</td>
			<td>Metrex</td>
			<td>11-1401</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Heparin Sodium 10,000 USP u/ml</td>
			<td>PPC</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Carprofen</td>
			<td>Metacam</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Glycopyrrolate</td>
			<td>Sandoz</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Isoflurane</td>
			<td>Abbott</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Chlohexidine 2%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Alcohol 70%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Iodine</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>PE-50</td>
			<td>BD</td>
			<td>427411</td>
			<td></td>
		 </tr>
		 <tr>
			<td>CO-EX T22</td>
			<td>Instech Solomon</td>
			<td>BCOEX-T22</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Connector 22G</td>
			<td>Instech Solomon</td>
			<td>SC22/15</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Swivel 22G</td>
			<td>Instech Solomon</td>
			<td>375/22PS</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Y-Connector 22G</td>
			<td>Instech Solomon</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Counterbalance and arm</td>
			<td>Instech Solomon</td>
			<td>CM375BP</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G blunted needles</td>
			<td>Instech Solomon</td>
			<td>LS23</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G canulation pins</td>
			<td>Instech Solomon</td>
			<td>SP23/12</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tethers (12 inch)</td>
			<td>Lomir</td>
			<td>RT12D</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Infusion jackets </td>
			<td>Lomir</td>
			<td>RJ01, RJ02, RJ03, RJ04</td>
			<td></td>
		 </tr>
		 <tr>
			<td>(SM-XL)</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tether attachment piece</td>
			<td>Lomir</td>
			<td>RS T1</td>
			<td></td>
		 </tr>
		 <tr>
			<td>60 ml syringe</td>
			<td>BD</td>
			<td>309653</td>
			<td></td>
		 </tr>
		 <tr>
			<td>1 ml syringe</td>
			<td>BD</td>
			<td>309602</td>
			<td></td>
		 </tr>
		</tbody>
 </table>

a model of chronic nutrient infusion in the rat

长期暴露在过多的营养水平推测影响多个器官和组织的功能和许多与肥胖相关的并发症，代谢综合征，包括2型糖尿病的发展做出贡献。要研究的机制，葡萄糖和脂肪酸的含量超标会影响胰腺β-细胞分泌胰岛素，我们已经建立了一种慢性营养输液在大鼠模型。该程序由右颈静脉和左颈总动脉置管全身麻醉下，允许7天的恢复期;导管连接使用一个旋转的泵和配重系统，使动物在笼子里自由移动;和注入葡萄糖和/或脂肪乳剂（大豆油乳剂与肝素注入时产生约80的％unsaturated/20％的饱和脂肪酸的混合物），72小时。该模型提供了一些优势的水电站，包括精细调节的目标水平的葡萄糖和脂肪酸循环的可能性;合作注入药类化合物的选项，以及相对短的时间内，而不是饮食模型。它可以用来检查营养引起的功能障碍的机制，在各种器官和测试药物的有效性，在此上下文中。

出一系列42只大鼠接受手术，在手术后期间丢失5只和1只，占整体86％的成功率在输液过程中丢失。最终注入的37只大鼠的平均体重为608±5克术前588±6克，开始输注（平均±SE，N = 37，P &lt;0.0001配对t检验）。 2输液组：生理盐水（SAL），葡萄糖+脂肪乳（GLU + IL）。 图2A和2B显示血糖和脂肪酸水平，分别获得以下具有代表性的结果，在72小时输液期间。按照设计，血糖水平维持?...

Watch this Scientific Journal Video about A Model of Chronic Nutrient Infusion in the Rat at JoVE.com

A Model of Chronic Nutrient Infusion in the Rat

慢性输注葡萄糖和脂肪乳大鼠的一种协议。该模型可用于研究营养过量对器官功能和生理参数的影响。

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

a-model-of-chronic-nutrient-infusion-in-the-rat

Research

JoVE Journal

Medicine

12.6K 次观看.  CRCHUM. 慢性输注葡萄糖和脂肪乳大鼠的一种协议。该模型可用于研究营养过量对器官功能和生理参数的影响。

视频: A Model of Chronic Nutrient Infusion in the Rat

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

膀胱胀诱发内脏运动反应为膀胱疼痛小鼠模型

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these diseases. Chronic retinal diseases such as retinal oedema or neovascularization usually require multiple intraocular injections to effectively treat the condition. However, the risks associated with these injections increase with repeated delivery of the drug. Therefore, alternative delivery methods need to be established in order to minimize the risks of reinjection. Several other investigations have developed methods to deliver drugs over extended time, through materials capable of releasing chemicals slowly into the eye. In this investigation, we outline the use of Elvax 40W, a copolymer resin, to act as a vehicle for drug delivery to the adult rat retina. The resin is made and loaded with the drug. The drug-resin complex is then implanted into the vitreous cavity, where it will slowly release the drug over time. This method was tested using 2-amino-4-phosphonobutyrate (APB), a glutamate analogue that blocks the light response of the retina. It was demonstrated that the APB was slowly released from the resin, and was able to block the retinal response by 7 days after implantation. This indicates that slow-release drug delivery using this copolymer resin is effective for treating the retina, and could be used therapeutically with further testing.

This paper details how Elvax 40W can be used as a slow-release method for drug delivery to the adult rat retina. The protocol for preparing, loading, and delivering the drug-resin complex to the eye is described.

通过40W的Elvax缓释给药的大鼠视网膜：对慢性疾病的治疗

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

Neurodegeneration in an Animal Model of Chronic Amyloid-beta Oligomer Infusion Is Counteracted by Antibody Treatment Infused with Osmotic Pumps

Decline in hippocampal-dependent explicit memory (memory for facts and events) is one of the earliest clinical symptom of Alzheimer&#39;s disease (AD). It is well established that synapse loss and ensuing neurodegeneration are the best predictors for memory impairments in AD. Latest studies have emphasized the neurotoxic role of soluble amyloid-beta oligomers (A&#946;o) that begin to accumulate in the human brain approximately 10 to 15 yr before the clinical symptoms become apparent. Many reports indicate that soluble A&#946;o correlate with memory deficits in AD models and humans. The A&#946;o-induced neurodegeneration observed in neuronal and brain slice cultures has been more challenging to reproduce in many animal models. The model of repeated A&#946;o infusions shown here overcome this issue and allow addressing two key domains for developing new disease modifying therapies: identify biological markers to diagnose early AD, and determine the molecular mechanisms underpinning A&#946;o-induced memory deficits at the onset of AD. Since soluble A&#946;o aggregate relatively fast into insoluble A&#946; fibrils that correlate poorly with the clinical state of patients, soluble A&#946;o are prepared freshly and injected once per day during six days to produce marked cell death in the hippocampus. We used cannula specially design for simultaneous infusions of A&#946;o and continuous infusion of A&#946;o antibody (6E10) in the hippocampus using osmotic pumps. This innovative in vivo method can now be used in preclinical studies to validate the efficiency of new AD therapies that might prevent the deposition and neurotoxicity of A&#946;o in pre-dementia patients.

To study the effects of A&#946;o in vivo, we developed a model based on repeated hippocampal infusions of soluble A&#946;o coupled with continuous infusion of A&#946;o antibody (6E10) in the hippocampus using osmotic pumps to counteract the neurotoxic effect of A&#946;o.

神经变性慢性淀粉样蛋白-β低聚物输注的动物模型是由抗体治疗灌注抵消与渗透泵

Decline in hippocampal-dependent explicit memory (memory for facts and events) is one of the earliest clinical symptom of Alzheimer&#39;s disease (AD). It ...

Use of a Central Venous Line for Fluids, Drugs and Nutrient Administration in a Mouse Model of Critical Illness

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting. Critically ill patients require intensive medical support in order to survive. While the majority of patients will recover within a few days, about a quarter of the patients need prolonged intensive care and are at high risk of dying from non-resolving multiple organ failure. Furthermore, the prolonged phase of critical illness is hallmarked by profound muscle weakness, and endocrine and metabolic changes, of which the pathogenesis is currently incompletely understood. The most widely used animal model in critical care research is the cecal ligation and puncture model to induce sepsis. This is a very reproducible model, with acute inflammatory and hemodynamic changes similar to human sepsis, which is designed to study the acute phase of critical illness. However, this model is hallmarked by a high lethality, which is different from the clinical human situation, and is not developed to study the prolonged phase of critical illness. Therefore, we adapted the technique by placing a central venous catheter in the jugular vein allowing us to administer clinically relevant supportive care, to better mimic the human clinical situation of critical illness. This mouse model requires an extensive surgical procedure and daily intensive care of the animals, but it results in a relevant model of the acute and prolonged phase of critical illness.

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting.

在重大疾病的小鼠模型中使用中心静脉导管的流体，药物和营养管理

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce ...

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 ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part of many experimental procedures where reagents need to be introduced directly into the bloodstream. Unwittingly, the injection can be subcutaneous, possibly altering the scientific outcomes. Utilizing a nanoemulsion-based biological probe with an incorporated near-infrared fluorescent (NIRF) dye, this method offers the capability of imaging a successful tail vein injection in vivo. With the use of a NIRF imager, images are taken before and after the injection of the agent. An acceptable IV injection is then qualitatively or quantitatively determined based on the intensity of the NIRF signal at the site of injection.

Here we present a method to validate tail vein injections in rats by utilizing near-infrared fluorescence imaging data from dyes incorporated into agents or biological probes. The tail is imaged before and after the injection, the fluorescent signal is quantified, and an assessment of the injection quality is made.

近红外标记剂对大鼠尾静脉注射的新的最佳验证

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...

Acupuncture in a Rat Model of Asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

哮喘鼠模型中的针灸

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

通过胆管 TNBS 输液治疗慢性胰腺炎的鼠标模型

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

改进大鼠模型中的 FSN 治疗：最大限度地减少不适并最大限度地提高效率

Efficacy of Fu's Subcutaneous Needling on Sciatic Nerve Pain: Behavioral and Electrophysiological Changes in a Chronic Constriction Injury Rat Model

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the mechanisms of action are still not fully understood. During FSN treatment, the FSN needle is inserted and retained in the subcutaneous tissues for a long duration with a swaying movement. However, challenges arise from maintaining a posture while manipulating FSN in animal models (e.g., rats) for researchers. Uncomfortable treatment can lead to fear and resistance to FSN needles, increasing the risk of injury and may even affect research data. Anesthesia may also affect the study results too. Hence, there is a need for strategies in FSN therapy on animals that minimize injury during the intervention. This study employs a chronic constriction injury model in Sprague-Dawley rats to induce neuropathic pain. This model replicates the pain induced by nerve injury observed in humans through surgical constriction of a peripheral nerve, mimicking the compression or entrapment seen in conditions such as nerve compression syndromes and peripheral neuropathies. We introduce an appropriate manipulation for easily inserting an FSN needle into the subcutaneous layer of the animal&#39;s body, including needle insertion and direction, needle retention, and swaying movement. Minimizing the rat&#39;s discomfort prevents the rat from being tense, which causes the muscle to contract and hinder the entry of the needle and improves the study efficiency.

作者聚焦：揭示 FSN 治疗的治疗效果 – 桥接神经性疼痛的研究和临床应用

We present a protocol for using Fu's subcutaneous needling in a chronic constriction injury model to induce sciatic nerve pain in rats.

傅氏皮下针刺治疗坐骨神经疼痛的疗效：慢性缩窄损伤大鼠模型中的行为和电生理变化

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the ...