我们要感谢来自辉瑞制药支持这项工作的合同（WS1763368）。我们还要感谢他们的支持沙特高等教育部（漠河）。

1。演习<ol><li>本实验的兴趣肌肘屈肌（肱二头肌）。 </li><li>肌力测定每个参与者能够给每一个人一个合适的电阻。这是由测试其电阻最大（RM）的每个参与者。 </li><li>为了测试的RM，我们用应变仪与计算机通过一个BIOPAC（DA - 100C）生物电放大器模块（BIOPAC系统，Goleta，CA）来衡量肌肉力量接口设备。该模块连接到MP - 100的模拟数字转换器的采样频率为1000赫兹每秒，分辨率24位（图10）。 </li><li>应变计装置固定在45度角的长椅。受试者被指示坐在后面的设备和休息，他们的胳膊肘上填充区域，使劳累的力量通过手腕。这是最好的方式，以确保这个问题不会吨招聘二头肌（图11）以外的任何肌肉。 </li><li>强度测定3次，每次收缩持续时间3秒分开约45秒的收缩。平均3个测量的RM。 </li><li>后，决定为每个参与者的二头肌的RM，旨在锻炼会议进行了35％的RM。 </li><li>所有科目都经历了同样的锻炼，使用适当的加权哑铃来诱导肌肉酸痛（DOMS）。这是做4套25重复二头肌浓度卷发，坐在椅子上，肘部与支持他们的大腿上（图12）。 </li><li>每个主题是每套之间休息90秒的时期。主题或全套25重复，或奉命停止，如果他们没有稳定控制在演习中的重量。 </li></ol> 2。红外线钙梅拉的制备​​及安装<ol><li>房间里的红外线成像发生在一个恒定的温度，以尽量减少任何在室温下的差异，这可能导致虚假的热读数的外部偏置。对于这个实验的目的，我们有一个控制房间温度保持在约23 ° C（+ / - 0.5 ° C）。 </li><li>相机被设置在1米以上的距离的距离，并在一个垂直的角度来看，皮肤被测量（图9A）* </li><li>成立所需的距离后，受试者被告知，直到图像已采取停滞不前。这不应该超过几秒钟，但它是非常关键的，以尽量减少运动，以确保拍摄的图像的准确性。 </li><li>这是最好的，房间里有较深颜色的油漆，而不是浅色，最大限度地减少任何红外线的干扰。 </li><li>照明也是很关键的处理与红外线图像，贝科时使用光源发出，像荧光灯或钨照明的红外线波可以提供虚假的高读数。最佳的照明选择是将一个房间配备了统一的LED灯，LED灯几乎不产生任何红外线的干扰（图9B）*. </li></ol> *使用FLIR 660红外摄像机（图8），在这里我们比较在不同的角度（0（垂直），15，30，45和60度）的皮肤图像在我们的实验室进行了一系列测试，并从皮肤（1，2，5米）在不同的距离，准确地检测皮肤的温度。所有的图像进行了比较，校准的热电偶，并在垂直角度和距离在1米距离的皮肤（R = 0.93）的图像和热电偶读数之间的相关性最好。不同的角度和距离造成的损失点阵化，减少之间的图像和热电偶读数的整体相关。 3。图片阿奎rement <ol><li>这个实验的目的，行使肌肉的形象是之前的练习，并在24和48小时后行使。 </li><li>目标比其他来源的身体热量可能会破坏热图像和提供虚假的读数。出于这个原因，任何人都不应站在旁边或背后的预定目标。 </li><li>在本次调查中，行使及非行使ARM的照片作比较。我们实行的武器之一，如前面提到的，和另一只手臂作为对照（图4和5）。 </li><li>从红外摄像机的图像数字被立即记录在一个单独的电子表格，因为它可能是难以确定属于谁的形象。 </li></ol> 4。图像处理与分析<ol><li>收购的红外图像处理使用“ThermoVision ExaminIR”软件版本：1.10.2。 </li><li>在选择所需的图像R分析，确定了四个地区的利益上获得的图像的手臂上使用的软件界面（图6）统计箱。 </li><li>当整个手臂已经找到所需的地区，该软件显示每个选定区域的温度的方法和标准偏差。然后，我们可以比较每个地区，无论是跨单独或获取整个手臂的平均气温（图7）。 </li></ol> 5。视觉类比量表及血液分析<ol><li>视觉模拟评分（VAS）来评估主观酸痛的手臂。规模（100毫米）一个10厘米长的行标“无痛苦”的一端，并在另一端的“非常疮”。每一名学员都沿10厘米的线的一个标志，以表明他们的反应，以酸胀。 </li><li>输精管的管理运动，运动后24小时，并在48小时前的科目。 </li><li>外周血是COL从学科lected，测量血液中的肌红蛋白的浓度水平。 </li><li>从受试者肘静脉血液是制定演习前，运动后30分钟结束，并在48小时内。 </li><li>血液离心10分钟4000转，从细胞中分离血清。样品随后被保存于-80℃直至做了肌红蛋白的分析。 </li><li>肌红蛋白测定采用一个TOSOH“友邦- 360”自动酶免疫分析仪（TOSOH公司，日本东京）。根据制造商的说明使用的肌红蛋白检测试剂盒（MYO 025297，ST友邦PACK肌红蛋白）。 </li></ol> 6。代表性的成果在本次调查采取的红外热图像的结果清楚地表示在图1。在3个时间段的41个科目的行​​使武器（运动前，运动后24小时，运动后48小时），拍摄的图像显示在温度显着增加，第2天（24小时运动后）时相比，运动前的温度，并在48小时内采取温度。正如图1所示，平均皮肤温度是32.80 + / - 1.03 ° C为1天（运动前）和33.96 + / - 1.46 ° C为2天（运动后24小时），32.82 + / - 第3天1.29（运动后48小时内）。这从1天到第2天皮肤温度差异有显着性意义（方差分析P &lt;0.01）。 然而，对于未行使的手臂，其中包括3个时间段的变化并不明显。图1显示，皮肤的平均温度是33.08 + / - 0.83 ° C为1天（运动前）和32.79 + / - 1.42 °彗星第2天（24小时运动后）和33.17 + / - 0.95第3天（运动后48小时内）。这在皮肤温度超过3天的差异不显著（方差分析P = 0.38）。 从VAS疼痛读数结果如图2所示。图图2，报告的疼痛已对2和第3天的急剧增加。行使的肌肉疼痛程度增加，从3.6 + / - 6.1第1天，36.3 + / - 2天的22.8和37.5 + / - 25.3 3天。从第1天的增加有显着性意义（方差分析P &lt;0.01）。 肌红蛋白的浓度水平的结果如图3所示。这个数字可以看出，第1天2肌红蛋白浓度（前，及30分钟后行使）之间几乎没有任何改变。但在第3天，在肌红蛋白的增加是非常大的。这3天增加约147微克每毫升血液（毫微克/毫升），较第1天第2浓度。肌红蛋白浓度分别为30.12 + / - 7.66 ng / mL的基线时，31.66 + / - 11.89 ng / mL的30分钟的运动后，和178.96 + / - 3天249.51纳克/毫升。这3天增加非常显著（方差分析P &lt;0.01）。 皮肤温度之间的相关分析得到的红外图像，与VAS疼痛水平。结果发现，有相当的相关性之间的第2天VAS读数，第2天皮肤温度测量。这种相关性是显着性（R = 0.312，P &lt;0.05）。然而，有没有明显的相关性之间的增值服务的读数，并在第3天的皮肤温度。这种相关性无显着相关（r = 0.047，P = 0.77）。 <img src="/files/ftp_upload/3551/3551fig1.jpg" alt="图1"/> 图1：一个代表在行使武器（钻石）的差异，在皮肤的温度，和行使联合国武器超过3天的时间内的41个科目（正方形）图。 <img src="/files/ftp_upload/3551/3551fig2.jpg" alt="图2"/> 图2：感知肌肉酸痛的测量差异与超过3天的时间内，所有的41个科目的的VAS的代表图。 图3一个代表超过3个时间段的所有41个科目中的肌红蛋白浓度差异图。 <img src="/files/ftp_upload/3551/3551fig4.jpg" alt="图4"/> 图4一）典型的一个主体行使 ARM的红外图像，然后再行使。 b）对同一科目手臂运动后24小时的红外图像。 <img src="/files/ftp_upload/3551/3551fig5.jpg" alt="图5"/> 图5一）典型的一个主题的联合国行使 ARM的红外图像，然后再行使。 b）对同一科目手臂运动后24小时的红外图像。 <img src="/files/ftp_upload/3551/3551fig6.jpg" alt="图6"/> 图6的4个地区的利益分析的THERM插图。人的手臂的形象。 <img src="/files/ftp_upload/3551/3551fig7.jpg" alt="图7"/> 图7。“ThermoVision ExaminIR”4箱利益行使手臂上显示红外图像化的软件界面。同时显示每个方块的统计解释。 <img src="/files/ftp_upload/3551/3551fig8.jpg" alt="图8"/> 图8。红外热成像摄像机，用于本次调查（FLIR 660） 。 <img src="/files/ftp_upload/3551/3551fig9.jpg" alt="图9"/> 图9）安装红外摄像机1米距离科目ARM。 B）的LED灯用于图像被送往实验室。 <img src="/files/ftp_upload/3551/3551fig10.jpg" alt="图10"/> 图10。）BIOPAC使用的模块，用于测量肌肉的力量。二）应变计装置固定到45 °角度工作台和挂接到BIOPAC系统。 <img src="/files/ftp_upload/3551/3551fig11.jpg" alt="图11"/> 图11：一个典型的主题上的应变计设备施加力。 <img src="/files/ftp_upload/3551/3551fig12.jpg" alt="图12"/> 图12：一个主题，接受诱导肌肉酸痛的运动协议。

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没有利益冲突的声明。

本次调查的主要目的是评估的热红外成像检测和测量，剧烈运动后肌肉酸痛的实用性，而且，我们的结果表明，红外成像可以为检测DOMS的有效技术，特别是在第一次24小时行使，。这并不奇怪，Pennes 18提供了一个非常详细的模型热流从肌肉四肢皮肤。该模型预测，在深层组织如肌肉的热量可以通过导电的热交换进入血液和皮肤进入消退。当肌肉运动，显然，巨大的热量是在肌肉中的肌纤?...

<table border="1"><tbody><tr><td> 该设备的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>红外线热成像摄像机</td><td> FLIR </td><td> FLIR SC660 </td><td></td></tr><tr><td>热红外分析软件</td><td> ThermoVision ExaminIR </td><td>软件版本1.10.2 </td><td></td></tr><tr><td>生物电放大器模块</td><td> BIOPAC </td><td> DA100C </td><td> DA100C提供可变增益设置和可调电压基准。 </td></tr><tr><td>模拟到数字转换器模块</td><td> BIOPAC </td><td> MP100 </td><td></td></tr><tr><td>自动酶免疫分析仪</td><td> TOSOH </td><td>友邦-360 </td><td>这个装置是用来分析血液样本，并获得了肌红蛋白的读数。 </td></tr>
 </tbody>
</table>

the use of thermal infra-red imaging to detect delayed onset muscle soreness

迟发性肌肉酸痛（DOMS），又称运动引起的肌肉损伤（EIMD），通常是曾长时间的身体活动，并开始练习1-4意外回合的个人经历，但也可以发生在运动员超出正常范围内行使他们的培训5。痛苦的现象与此相关的症状范围从轻微的肌肉压痛，严重衰弱疼痛 1,3,5 。这些症状和相关的不适，增加第一个24小时内终止行使，及24至72小时后练习1,3之间的峰的强度。出于这个原因，DOMS的是经常运动损伤最常见的形式，可以影响一个人的表现，并成为许多1,4恐吓之一。 在过去的三十年，DOMS的现象已经获得了相当数量的其中包括研究人员和专家，在运动生理学，体育，和康复领域 6的兴趣。已有各种调查这种痛苦发生在其潜在的机制，治疗干预，并1-5,7-12预防战略方面的研究发表。然而，这是显而易见的，从文学，DOMS不是一件容易的的病理量化，因为有一个变异之间的测量工具和方法，用来量化这种情况下6。这是显而易见的，一个最好的评价措施DOMS的，这使得它难以核实是否在一个特定的干预确实有助于减少这种类型的疼痛或不相关的症状一直没有达成协议。因此，可以看出DOMS的有点含糊，因为许多研究取决于测量疼痛采用视觉模拟评分（VAS）10,13-15，这是一种主观的，而不是一个客观的衡量。即使针肌肉活检，myofibre蛋白质的血药浓度可能会考虑一些大的变化，在这些血蛋白 6金标准已记录在 6,16，除了有时与微创技术相关的高风险。 因此，在目前的调查中，我们测试了皮肤的热红外（IR）成像行使肌肉以上技术检测相关的肌肉酸痛。红外热成像已被使用，被发现在自1950年的17个不同类型的疾病和感染检测成功。但令人惊讶的是，附近没有完成DOMS和皮肤温度的变化。本次调查的主要目的是使用这种安全和非侵入性技术研究在DOMS的变化。

Watch this Scientific Journal Video about The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness at JoVE.com

The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

本次调查的目的是评估是否使用红外热成像摄像机是一个有效的检测和定量运动后肌肉酸痛工具。

热红外成像来检测迟发性肌肉酸痛

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically ...

the-use-thermal-infra-red-imaging-to-detect-delayed-onset-muscle

Research

JoVE Journal

Medicine

19.1K 次观看.  Loma Linda University. 本次调查的目的是评估是否使用红外热成像摄像机是一个有效的检测和定量运动后肌肉酸痛工具。

视频: The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Thermal Ablation for the Treatment of Abdominal Tumors

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ablation procedure, a thin applicator is guided into the target tumor under imaging guidance. Energy is then applied to the tissue until temperatures rise to cytotoxic levels (50-60 &deg;C). Various energy sources are available to heat biological tissues, including radiofrequency (RF) electrical current, microwaves, laser light and ultrasonic waves. Of these, RF and microwave ablation are most commonly used worldwide.
During RF ablation, alternating electrical current (~500 kHz) produces resistive heating around the interstitial electrode. Skin surface electrodes (ground pads) are used to complete the electrical circuit. RF ablation has been in use for nearly 20 years, with good results for local tumor control, extended survival and low complication rates1,2. Recent studies suggest RF ablation may be a first-line treatment option for small hepatocellular carcinoma and renal-cell carcinoma3-5. However, RF heating is hampered by local blood flow and high electrical impedance tissues (eg, lung, bone, desiccated or charred tissue)6,7. Microwaves may alleviate some of these problems by producing faster, volumetric heating8-10. To create larger or conformal ablations, multiple microwave antennas can be used simultaneously while RF electrodes require sequential operation, which limits their efficiency. Early experiences with microwave systems suggest efficacy and safety similar to, or better than RF devices11-13.
Alternatively, cryoablation freezes the target tissues to lethal levels (-20 to -40 &deg;C). Percutaneous cryoablation has been shown to be effective against RCC and many metastatic tumors, particularly colorectal cancer, in the liver14-16. Cryoablation may also be associated with less post-procedure pain and faster recovery for some indications17. Cryoablation is often contraindicated for primary liver cancer due to underlying coagulopathy and associated bleeding risks frequently seen in cirrhotic patients. In addition, sudden release of tumor cellular contents when the frozen tissue thaws can lead to a potentially serious condition known as cryoshock 16.
Thermal tumor ablation can be performed at open surgery, laparoscopy or using a percutaneous approach. When performed percutaneously, the ablation procedure relies on imaging for diagnosis, planning, applicator guidance, treatment monitoring and follow-up. Ultrasound is the most popular modality for guidance and treatment monitoring worldwide, but computed tomography (CT) and magnetic resonance imaging (MRI) are commonly used as well. Contrast-enhanced CT or MRI are typically employed for diagnosis and follow-up imaging.

A thermal tumor ablation procedure is described. The entire procedure is detailed, including pretreatment planning and imaging studies, anesthesia, adjuvant techniques to facilitate a percutaneous approach, imaging guidance of the ablation device to the tumor, thermal treatment, post-treatment care and follow-up.

热消融治疗腹部肿瘤

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ...

科研

医学

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment for melanoma remains surgical excision, therefore, the key to extended survival is early detection 2,3. Considering the large numbers of patients diagnosed every year and the limitations in accessing specialized care quickly, the development of objective in vivo diagnostic instruments to aid the diagnosis is essential. New techniques to detect skin cancer, especially non-invasive diagnostic tools, are being explored in numerous laboratories. Along with the surgical methods, techniques such as digital photography, dermoscopy, multispectral imaging systems (MelaFind), laser-based systems (confocal scanning laser microscopy, laser doppler perfusion imaging, optical coherence tomography), ultrasound, magnetic resonance imaging, are being tested. Each technique offers unique advantages and disadvantages, many of which pose a compromise between effectiveness and accuracy versus ease of use and cost considerations. Details about these techniques and comparisons are available in the literature 4.
Infrared (IR) imaging was shown to be a useful method to diagnose the signs of certain diseases by measuring the local skin temperature. There is a large body of evidence showing that disease or deviation from normal functioning are accompanied by changes of the temperature of the body, which again affect the temperature of the skin 5,6. Accurate data about the temperature of the human body and skin can provide a wealth of information on the processes responsible for heat generation and thermoregulation, in particular the deviation from normal conditions, often caused by disease. However, IR imaging has not been widely recognized in medicine due to the premature use of the technology 7,8 several decades ago, when temperature measurement accuracy and the spatial resolution were inadequate and sophisticated image processing tools were unavailable. This situation changed dramatically in the late 1990s-2000s. Advances in IR instrumentation, implementation of digital image processing algorithms and dynamic IR imaging, which enables scientists to analyze not only the spatial, but also the temporal thermal behavior of the skin 9, allowed breakthroughs in the field.
In our research, we explore the feasibility of IR imaging, combined with theoretical and experimental studies, as a cost effective, non-invasive, in vivo optical measurement technique for tumor detection, with emphasis on the screening and early detection of melanoma 10-13. In this study, we show data obtained in a patient study in which patients that possess a pigmented lesion with a clinical indication for biopsy are selected for imaging. We compared the difference in thermal responses between healthy and malignant tissue and compared our data with biopsy results. We concluded that the increased metabolic activity of the melanoma lesion can be detected by dynamic infrared imaging.

We demonstrated that malignant pigmented lesions with increased metabolic activity generate quantifiable amounts of heat and the measurement of the transient thermal response of the skin to a cooling excitation allows quantitative identification of melanoma and other skin cancers (vs. non-proliferative nevi) at an early stage of the disease.

定量可视化和皮肤癌症使用动态热成像检测

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

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

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

脓毒症动物模型，用于评估新型草药疗法

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

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

挑战的药理功能磁共振成像：在临床前研究使用的5  - 羟色胺系统中的应用

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle degeneration1, 2. In patients, the ability of resident muscle satellite cells (SCs) to regenerate damaged myofibers becomes increasingly inefficient4. Therefore, transplantation of muscle progenitor cells (MPCs)/myoblasts from healthy subjects is a promising therapeutic approach to DMD. A major limitation to the use of stem cell therapy, however, is a lack of reliable imaging technologies for long-term monitoring of implanted cells, and for evaluating its effectiveness. Here, we describe a non-invasive, real-time approach to evaluate the success of myoblast transplantation. This method takes advantage of a unified fusion reporter gene composed of genes (firefly luciferase [fluc], monomeric red fluorescent protein [mrfp] and sr39 thymidine kinase [sr39tk]) whose expression can be imaged with different imaging modalities9, 10. A variety of imaging modalities, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), optical imaging, and high frequency 3D-ultrasound are now available, each with unique advantages and limitations11. Bioluminescence imaging (BLI) studies, for example, have the advantage of being relatively low cost and high-throughput. It is for this reason that, in this study, we make use of the firefly luciferase (fluc) reporter gene sequence contained within the fusion gene and bioluminescence imaging (BLI) for the short-term localization of viable C2C12 myoblasts following implantation into a mouse model of DMD (muscular dystrophy on the X chromosome [mdx] mouse)12-14. Importantly, BLI provides us with a means to examine the kinetics of labeled MPCs post-implantation, and will be useful to track cells repeatedly over time and following migration. Our reporter gene approach further allows us to merge multiple imaging modalities in a single living subject; given the tomographic nature, fine spatial resolution and ability to scale up to larger animals and humans10,11, PET will form the basis of future work that we suggest may facilitate rapid translation of methods developed in cells to preclinical models and to clinical applications.

A non-invasive means to evaluate the success of myoblast transplantation is described. The method takes advantage of a unified fusion reporter gene composed of genes whose expression can be imaged with different imaging modalities. Here, we make use of a fluc reporter gene sequence to target cells via bioluminescence imaging.

目标移植的肌肉祖细胞的分子成像

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

高通量流式细胞仪检测细胞为基础的试验检测抗体，以N-甲基-D-天冬氨酸受体或多巴胺2受体在人血清

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

小鼠模型的非侵入性成像检测和监测卵巢癌复发

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of living systems. Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using qMRI methods. The MRI acquisition protocol begins with localizer images (used to locate the position of the body and tissue of interest within the MRI system), quality control measurements of relevant magnetic field distributions, and structural imaging for general anatomical characterization. The qMRI portion of the protocol includes measurements of the longitudinal and transverse relaxation time constants (T1 and T2, respectively). Also acquired are diffusion-tensor MRI data, in which water diffusivity is measured and used to infer pathological processes such as edema. Quantitative magnetization transfer imaging is used to characterize the relative tissue content of macromolecular and free water protons. Lastly, fat-water MRI methods are used to characterize fibro-adipose tissue replacement of muscle. In addition to describing the data acquisition and analysis procedures, this paper also discusses the potential problems associated with these methods, the analysis and interpretation of the data, MRI safety, and strategies for artifact reduction and protocol optimization.

Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using non-invasive magnetic resonance imaging methods.

骨骼肌疾病的定量磁共振成像

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

主动脉钙化和炎症血管平滑肌细胞和成像的钙化

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include supporting mastication and deglutition, as well as taste-sensing and phonetics. Accordingly, the strength and volume of the tongue can impact the ability of vertebrates to accomplish basic activities such as feeding, communicating, and breathing. Human patients with sleep apnea have enlarged tongues, characterized by reduced muscle tone and increased intramuscular fat that can be visualized and quantified by magnetic resonance imaging (MRI). The abilities to measure force generation and viscoelastic properties of the tongue constitute important tools for obtaining functional information to correlate with imaging data. Here, we present techniques for measuring tongue force production in anesthetized Zucker rats via electrical stimulation of the hypoglossal nerves and for determining the viscoelastic properties of the tongue by applying passive Lissajous force/deformation curves.

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

We describe a surgical procedure in an anesthetized rat model for determining the muscle tone and viscoelastic properties of the tongue. The procedure involves specific stimulation of the hypoglossal nerves and application of passive Lissajous force/deformation curves to the muscle.