1。动物伦理在执行长持续时间热疗对小鼠确保机构动物管理和使用委员会已批准的程序。本文讨论的所有程序，并显示在影随行的国家免疫学研究所实验动物伦理委员会的批准。 2。了解热疗全身热疗（WBH）被传递放置在预热的网箱孵化器被放置在保持在39.5℃下的动物在标准化的协议，一个温度传感器，在小鼠体内的外科手术植入，并在不同的实验条件下监测其体温。据观察，在热环境下的动物时，它们的温度逐渐上升至37°C到42°C，持续时间为100分钟（ 图1）。钍Ë产生的“秒杀”的温度是耐人寻味的，因为身体内部的温度射门甚至高于外部环境温度。 当身体温度超过40℃，动物将显示如润湿口鼻部周围的应力和脱水的迹象。增加身体的温度超过40°C，会导致润湿性和不可逆的压力，甚至死亡的动物，由于热应力。因此，重要的是要防止在热疗过程中的温度峰值。 这种高温尖峰现象关联与物理亢进的动物的在热疗曝光期间。相关温度峰值由其他组中观察到过，并与的动物，在被引入到环境中具有更高的温度下的10的物理多动症。 在我们的协议中，小鼠被固定在热疗曝光。如如图2所示，固定化的动物的防止温度峰值。 我们用8-12周龄BALB / cJ小鼠，证明此协议。该协议并不限于一个特定的类型/应变的小鼠。 3。实验准备 3.1孵化器该实验可在一个正常的CO 2培养箱中进行，因为它们具有良好的温度控制。断开CO 2气体的供给和除去水的CO 2培养箱中保持托盘。在某些型号的孵化器中，它是可以关掉的CO 2控制器。安排货架上的孵化器，一个老鼠笼可以方便放在里面的。孵化器的温度设定在39.5℃。 3.2网箱通常，热疗进行实验，在一个时间上的4只小鼠。两笼;e被置于孵化器内的，另一种是在室温下放在外面。床上用品的笼子是可以避免的，因为随着时间的推移，这将导致非均匀温度在笼子里面的床上用品变湿的补丁。外面的笼子应该有食物颗粒和一瓶水。 3.3小鼠固定管在热疗小鼠的第4个周期的过程中被固定在穿孔管。可以由这些固定管50 ML&#39;离心机&#39;的管，通过穿孔3毫米的孔，​​所有超过它的身体和将需要四个这种管子。 50毫升管可以方便的固定BALB / cJ小鼠，年龄超过8周。管可能需要适当大小的小鼠或一些不寻常的不同菌株，这是非常年轻的年龄超过50周的小鼠。 将固定管和孵化器内的一个笼子里保持在39.5°C。等待至少2个小时，所以笼子里，我mmobilization管的孵化器中达到的温度。 4。第一周期的热疗小鼠可能会​​遇到脱水在热疗曝光。因此，在实验开始之前，管理0.5 ml生理盐水腹腔内的每一个鼠标。使用26 G针1毫升注射器。 的笼子放置在孵化器中迅速取出，放置在每个固定管一个动物的笼子放置在孵化器背面。记录时间。关键是要保持笼子的孵化器每次在同一个地方。 90分钟后，取出笼从孵化器。松开固定管的动物，并把它们在笼子里，在常温下容易获得食物和水。将空的固定管的孵化器。 15分钟后，在第二个周期开始的热疗通过提供0。5 ml生理盐水腹腔内的每一个鼠标。 然后将它们固定在50毫升的穿孔固定管上，并放置在孵化器中。 5。随后的热疗周期对于第4个周期，它是必不可少的固定小鼠在热疗曝光期间的前两个周期，盐水注射热疗期间开始之前。 这些周期后，休息热疗完成后，小鼠获得适应环境并没有变得异常活跃的热疗过程中暴露。腹腔注射生理盐水可能没有必要，如果老鼠随手拿食物和水的休息时间。 6。完成的周期后，最后一步是检查动物不应该有任何的死亡在热疗周期。每种动物都有非常仔细地观看第一几个周期。快速查看任何润湿周围的吻。这种症状是脱水的签名。小鼠有此类症状，应立即给予0.5毫升生理盐水ip和其他在室温下。大多数的固定小鼠不需要任何额外的照顾，并会成功地承受压力的热疗。 完成所需的周期后，动物可能看起来虚弱和脱水，但他们会在几个小时内迅速恢复。 <img src="/files/ftp_upload/3801/3801fig1.jpg" alt="图1" /> 图1小鼠的培养箱中于39.5℃，当笼置于芯体温度变化笼90分钟后，从培养箱中取出，并放置在25℃。红色圆圈指示高的温度峰值。 <img src="/files/ftp_upload/3801/3801fig2.jpg" alt="图2" /> 芯体temperatu变化如图2所示。重新热疗周期期间的固定化的小鼠。固定动物后，高温峰值时不遵守笼被放置在39.5°C。

<ol>
	<li>Braude, A. I., Beck, J., Zalesky, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Febrile+response+to+bacterial+pyrogens+in+leukemia.">Febrile response to bacterial pyrogens in leukemia.</a> Blood. 16, 1279 (1960).</li><li>Dovern, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperthermic+intraperitoneal+chemotherapy+added+to+the+treatment+of+ovarian+cancer.+A+review+of+achieved+results+and+complications.">Hyperthermic intraperitoneal chemotherapy added to the treatment of ovarian cancer. A review of achieved results and complications.</a> Eur. J. Gynaecol. Oncol. 31, 256 (2010).</li><li>Gaworek, J., Mayer, C. 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我们什么都没有透露。

<html>早在5000 BC，，埃及医生治疗肿瘤热。希腊人认识到，在一些医疗护理的热值。最古老的文本，摩西提到的“温泉（创世记36:24），，治疗提高身体的温度。 早期的技术使用气相微生物提取物诱导发热或热水浴诱导热疗1。然而，这些方法可以很容易地监视或控制。在我们的协议小鼠暴露于高热的控制和可重复的方式。 在此过程中，重要的是对...

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protocol for long duration whole body hyperthermia in mice

热疗是一个通用术语，用来定义在核心体温高于正常的增加。它经常被用于描述增加芯体的温度的过程中观察到发热。热疗作为佐剂的使用已经出现作为一个有前途的癌症生物学领域中的肿瘤消退的过程。用于此目的，最重要的要求是具有可靠和均匀的加热协议。我们已经开发出了一个协议，用于热疗（全身）的小鼠。在这个协议中，动物被暴露于高温的周期为90分钟，然后通过15分钟的休息时间。在此期间，小鼠很容易获得食物和水。在最初的几个的热疗曝光周期的小鼠体温高尖峰，防止固定的动物。此外，施用生理盐水前几个周期中的脱水的影响减到最小。这个协议可以模拟条件下的小鼠12-24小时出现发烧一样。我们有使用8-12周龄BALB / Cj的雌性小鼠证明协议。

Watch this Scientific Journal Video about Protocol for Long Duration Whole Body Hyperthermia in Mice at JoVE.com

Protocol for Long Duration Whole Body Hyperthermia in Mice

本文介绍了一种协议，用于全身热疗的小鼠发烧一样，可以刺激条件下可达12-24小时。

持续时间较长的小鼠全身热疗协议

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

protocol-for-long-duration-whole-body-hyperthermia-in-mice

Research

JoVE Journal

Medicine

11.5K 次观看.  National Institute of Immunology. 本文介绍了一种协议，用于全身热疗的小鼠发烧一样，可以刺激条件下可达12-24小时。

视频: Protocol for Long Duration Whole Body Hyperthermia in Mice

Protocols for Assessing Radiofrequency Interactions with Gold Nanoparticles and Biological Systems for Non-invasive Hyperthermia Cancer Therapy

Cancer therapies which are less toxic and invasive than their existing counterparts are highly desirable. The use of RF electric-fields that penetrate deep into the body, causing minimal toxicity, are currently being studied as a viable means of non-invasive cancer therapy. It is envisioned that the interactions of RF energy with internalized nanoparticles (NPs) can liberate heat which can then cause overheating (hyperthermia) of the cell, ultimately ending in cell necrosis.
In the case of non-biological systems, we present detailed protocols relating to quantifying the heat liberated by highly-concentrated NP colloids. For biological systems, in the case of in vitro experiments, we describe the techniques and conditions which must be adhered to in order to effectively expose cancer cells to RF energy without bulk media heating artifacts significantly obscuring the data. Finally, we give a detailed methodology for in vivo mouse models with ectopic hepatic cancer tumors.

We describe the protocols used to investigate the interactions of 13.56 MHz radiofrequency (RF) electric-fields with gold nanoparticle colloids in both non-biological and biological systems (in vitro/vivo). These interactions are being investigated for applications in cancer therapy.

协议的射频评估与互动金纳米粒子和生物系统的非侵入性癌症热疗治疗

Cancer therapies  which are less toxic and invasive than their existing counterparts are highly  desirable. The use of RF electric-fields that penetrate ...

科研

医学

Parabiosis in Mice: A Detailed Protocol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 18641, the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival2. In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

联体小鼠：一个详细的协议

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Long-term Blood Pressure Measurement in Freely Moving Mice Using Telemetry

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. Indeed, research focusing on the discovery of new potential therapeutic targets using genetically altered mice requires a reliable, long-term assessment of the systemic arterial pressure variation. Currently, the gold standard for obtaining long-term measurements of blood pressure in ambulatory mice uses implantable radio-transmitters, which require&#160;artery cannulation. This technique eliminates the need for tethering, restraining, or anesthetizing the animals which introduce stress and artifacts during data sampling. However, arterial blood pressure monitoring in mice via catheterization can be rather challenging due to the small size of the arteries. Here we present a step-by-step guide to illustrate the crucial key passages for a successful subcutaneous implantation of radio-transmitters and carotid artery cannulation in mice. We also include examples of long-term blood pressure activity taken from freely moving mice after a period of post-surgery recovery. Following this procedure will allow reliable direct blood pressure recordings from multiple animals simultaneously.

The goal of this protocol is to assess systemic blood pressure in conscious freely moving mice using implantable radio-telemetry devices.

自由活动小鼠使用遥测长期血压测量

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. ...

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

高脂喂养小鼠体内成分及代谢锁定分析

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

Protocol to Create Chronic Wounds in Diabetic Mice

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ~6.5M people and cost ~$40B/year in the US alone. Although a significant effort has been invested in understanding how chronic wounds develop in humans, fundamental questions remain unanswered. Recently, we developed a novel mouse model for diabetic chronic wounds that have many characteristics of human chronic wounds. Using db/db-/- mice, we can generate chronic wounds by inducing high levels of oxidative stress (OS) in the wound tissue immediately after wounding, using a one-time treatment with inhibitors specific to the antioxidant enzymes catalase and glutathione peroxidase. These wounds have high levels of OS, develop biofilm naturally, become fully chronic within 20 days after treatment and can remain open more for more than 60 days. This novel model has many features of diabetic chronic wounds in humans and therefore can contribute significantly to advancing fundamental understanding of how wounds become chronic. This is a major breakthrough because chronic wounds in humans cause significant pain and distress to patients and result in amputation if unresolved. Moreover, these wounds are very expensive and time-consuming to treat, and lead to significant loss of personal income to patients. Advancements in this field of study through the use of our chronic wound model can significantly improve health care for millions who suffer under this debilitating condition. In this protocol, we describe in great detail the procedure to cause acute wounds to become chronic, which has not been done before.

Chronic wounds are developed from acute wounds on a diabetic mouse model by inducing high levels of oxidative stress after a full-thickness cutaneous wound. The wound is treated with inhibitors for catalase and glutathione peroxidase, resulting in impaired healing and biofilm development by bacteria present in the skin microbiome.

在糖尿病小鼠中创建慢性伤口的协议

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ...

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a handheld metal detector to detect ingested metallic foreign bodies in children. We propose that by introducing handheld metal detector screening early in the triage process of children with high suspicion of metallic foreign body ingestion, the number of radiographs being ordered to localize the metallic foreign body can be reduced in this radio-sensitive population. The study protocol requires the screening of the participants for history of foreign body ingestion and exclusion of patients with respiratory distress or metallic implants. The patient changes to hospital gown and items that could contain metal like eyeglasses, earrings, pendants, and ornaments are removed. The patient is positioned in the center of the room away from other metallic interferences. The working status of the handheld metal detector is first confirmed by eliciting a positive audio-visual signal. Then the screening is done in an erect position with head in extension to expose the neck, from the level of the chin to the level of the hip joint, to cover the anatomical areas from neck to pelvis in a zig-zag manner both anteriorly and posteriorly. A positive audio-visual signal is carefully noted during the scanning for the presence of metallic foreign body. Relevant radiographs are ordered as per the area detected on the metal detector screening. The handheld metal detector was able to precisely identify all the coins among the ingested metallic foreign bodies in our study. The handheld metal detector could not consistently detect non-coin metallic foreign bodies. This protocol demonstrates the accuracy of handheld metal detector in the identification and localization of coins and coin like metallic foreign bodies.

Here, we present a protocol to use the handheld metal detector, to screen for the presence of ingested metallic foreign bodies in children who present to the pediatric emergency medicine department with a history of foreign body ingestion.

金属异物在儿童体内摄取的手持金属探测器筛选

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

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy (PRRT): 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods

Peptide-receptor-radionuclide-therapy (PPRT) is a targeted therapy that combines a short-range energy radionuclide with a substrate with high specificity for cancer cell receptors. After injection, the radiotracer is distributed throughout the entire body, with a higher uptake in tissues where targeted receptors are overexpressed. The use of beta/gamma radionuclide emitters enables therapy imaging (beta-emission) and post-therapy imaging (gamma-emission) to be performed at the same time. Post-treatment sequential images permit absorbed dose calculation based on local uptake and wash-in/wash-out kinetics. We implemented a hybrid method that combines information derived from both 2D and 3D images. Serial whole-body images and blood samples are acquired to estimate the absorbed dose to different organs at risk and to lesions disseminated throughout the body. A single 3D-SPECT/CT image, limited to the abdominal region, overcomes projection overlap on planar images of different structures such as the intestines and kidneys. The hybrid 2D+3D-SPECT/CT method combines the effective half-life information derived from 2D planar images with the local uptake distribution derived from 3D images. We implemented this methodology to estimate the absorbed dose for patients undergoing PRRT with 177Lu-PSMA-617. The methodology could, however, be implemented with other beta-gamma radiotracers. To date, 10 patients have been enrolled into the dosimetry study with 177Lu-PSMA-617 combined with drug protectors for kidneys and salivary glands (mannitol and glutamate tablets, respectively). The median ratio between kidney uptake at 24 h evaluated on planar images and 3D-SPECT/CT is 0.45 (range:0.32-1.23). The comparison between hybrid and full 3D approach has been tested on one patient, resulting in a 1.6% underestimation with respect to full 3D (2D: 0.829 mGy/MBq, hybrid: 0.315 mGy/MBq, 3D: 0.320 mGy/MBq). Treatment safety has been confirmed, with a mean absorbed dose of 0.73 mGy/MBq (range:0.26-1.07) for kidneys, 0.56 mGy/MBq (0.33-2.63) for the parotid glands and 0.63 mGy/MBq (0.23-1.20) for submandibular glands, values in accordance with previously published data.

A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy PRRT: 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods

This method estimates the absorbed dose of different structures for peptide-receptor-radionuclide-therapy (PRRT) with the possibility of avoiding organ overlap on 2D-projections. Serial whole-body planar images permit estimation of mean absorbed doses along the whole body, while the hybrid approach, combining planar images and 3D-SPECT/CT image, overcomes the limitations of structure overlapping.

肽受体放射性核素治疗（PRRT）全身多体型测定方案：2D平面图像和混合2D+3D SPECT/CT图像方法

Peptide-receptor-radionuclide-therapy (PPRT) is a targeted therapy that combines a short-range energy radionuclide with a substrate with high specificity ...

Assessing Whole-Body Lipid-Handling Capacity in Mice

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a class of many different molecules with many pathways involved in their synthesis and metabolism. A starting point for evaluating lipid hemostasis for nutrition and obesity research is needed. This paper describes three easy and accessible methods that require little expertise or practice to master, and that can be adapted by most labs to screen for lipid-metabolism abnormalities in mice. These methods are (1) measuring several fasting serum lipid molecules using commercial kits (2) assaying for dietary lipid-handling capability through an oral intralipid tolerance test, and (3) evaluating the response to a pharmaceutical compound, CL 316,243, in mice. Together, these methods will provide a high-level overview of lipid handling capability in mice.

This paper provides three easy and accessible assays for assessing lipid metabolism in mice.

评估小鼠全身脂质处理能力

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a ...