这项研究是由国立卫生研究院（R21 MH090384，R01 EB006494，RO1 EB009666，R01 NS051622）资助。 H. Sarofin C. Lacadie我们感谢他们的技术援助。

<ol>
	<li>Mataix-Cols, D., Cullen, S., Lange, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+correlates+of+anxiety+associated+with+obsessive-compulsive+symptom+dimensions+in+normal+volunteers.">Neural correlates of anxiety associated with obsessive-compulsive symptom dimensions in normal volunteers.</a> Biol. Psychiatry. 53, 482-493 (2003).</li><li>Mataix-Cols, D., Wooderson, S., Lawrence, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+neural+correlates+of+washing,+checking,+and+hoarding+symptom+dimensions+in+obsessive-compulsive+disorder.">Distinct neural correlates of washing, checking, and hoarding symptom dimensions in obsessive-compulsive disorder.</a> Arch. Gen. Psychiatry. 61, 564-576 (2004).</li><li>Menzies, L., Chamberlain, S. R., Laird, A. 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M., Fairneny, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+cortical+activity+in+auditory+areas+through+neurofeedback+functional+magnetic+resonance+imaging.">Increasing cortical activity in auditory areas through neurofeedback functional magnetic resonance imaging.</a> Neuroreport. 17, 1273-1278 (2006).</li><li>Caria, A., Sitaram, R., Veit, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volitional+control+of+anterior+insula+activity+modulates+the+response+to+aversive+stimuli.+A+real-time+functional+magnetic+resonance+imaging+study.">Volitional control of anterior insula activity modulates the response to aversive stimuli. 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E., Healy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Upregulation+of+emotion+areas+through+neurofeedback+with+a+focus+on+positive+mood.">Upregulation of emotion areas through neurofeedback with a focus on positive mood.</a> Cognitive, affective &#38; behavioral neuroscience. 11, 44-51 (2011).</li><li>Johnston, S. J., Boehm, S. G., Healy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurofeedback:+A+promising+tool+for+the+self-regulation+of+emotion+networks.">Neurofeedback: A promising tool for the self-regulation of emotion networks.</a> NeuroImage. 49, 1066-1072 (2010).</li><li>Zotev, V., Krueger, F., Phillips, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-regulation+of+amygdala+activation+using+real-time+fMRI+neurofeedback.">Self-regulation of amygdala activation using real-time fMRI neurofeedback.</a> PLoS One. 6, e24522-e24522 (2011).</li><li>Haller, S., Birbaumer, N., Veit, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+fMRI+feedback+training+may+improve+chronic+tinnitus.">Real-time fMRI feedback training may improve chronic tinnitus.</a> Eur. Radiol. 20, 696-703 (2010).</li><li>Bloch, M. H., Landeros-Weisenberger, A., Kelmendi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review:+antipsychotic+augmentation+with+treatment+refractory+obsessive-compulsive+disorder.">A systematic review: antipsychotic augmentation with treatment refractory obsessive-compulsive disorder.</a> Mol. Psychiatry. 11, 622-632 (2006).</li><li>Jenike, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+practice.+Obsessive-compulsive+disorder.">Clinical practice. Obsessive-compulsive disorder.</a> N. Engl. J. Med. 350, 259-265 (2004).</li><li>Pallanti, S., Quercioli, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment-refractory+obsessive-compulsive+disorder:+methodological+issues,+operational+definitions+and+therapeutic+lines.">Treatment-refractory obsessive-compulsive disorder: methodological issues, operational definitions and therapeutic lines.</a> Prog. Neuropsychopharmacol. Biol. Psychiatry. 30, 400-412 (2006).</li><li>Mataix-Cols, D., Lawrence, N. S., Wooderson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Maudsley+Obsessive-Compulsive+Stimuli+Set:+validation+of+a+standardized+paradigm+for+symptom-specific+provocation+in+obsessive-compulsive+disorder.">The Maudsley Obsessive-Compulsive Stimuli Set: validation of a standardized paradigm for symptom-specific provocation in obsessive-compulsive disorder.</a> Psychiatry. Res. 168, 238-241 (2009).</li><li>Lang, P. J., Bradley, M. M., Cuthbert, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=International+affective+picture+system+(IAPS):+Affective+ratings+of+pictures+and+instruction+manual.">International affective picture system (IAPS): Affective ratings of pictures and instruction manual.</a> Technical Report A-82008. , (2008).</li><li>Burns, G. L., Keortge, S. G., Formea, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revision+of+the+Padua+Inventory+of+obsessive+compulsive+disorder+symptoms:+distinctions+between+worry,+obsessions,+and+compulsions.">Revision of the Padua Inventory of obsessive compulsive disorder symptoms: distinctions between worry, obsessions, and compulsions.</a> Behaviour research and therapy. 34, 163-173 (1996).</li><li>Scheinost, D., Hampson, M., Bhawnani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+GPU+accelerated+motion+correction+algorithm+for+real-time+fMRI.">A GPU accelerated motion correction algorithm for real-time fMRI.</a> Human Brain Mapping. , 639 (2011).</li><li>Hampson, M., Scheinost, D., Qiu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biofeedback+from+the+supplementary+motor+area+reduces+functional+connectivity+to+subcortical+regions.">Biofeedback from the supplementary motor area reduces functional connectivity to subcortical regions.</a> Brain Connectivity. 1, 91-98 (2011).</li><li>Johnson, K. A., Hartwell, K., Lematty, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+"Real-time"+fMRI+Feedback+Is+Superior+to+Continuous+Presentation+for+a+Motor+Imagery+Task:+A+Pilot+Study.">Intermittent "Real-time" fMRI Feedback Is Superior to Continuous Presentation for a Motor Imagery Task: A Pilot Study.</a> J. Neuroimaging. , (2011).</li><li>Yoo, S. S., Lee, J. H., O'Leary, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+magnetic+resonance+imaging-mediated+learning+of+increased+activity+in+auditory+areas.">Functional magnetic resonance imaging-mediated learning of increased activity in auditory areas.</a> Neuroreport. 18, 1915-1920 (2007).</li></ol>

没有利益冲突的声明。

实时fMRI数据的生物反馈是一种新技术，需要更多的工作来优化这种方法，以最大限度地提高学习的科目。最近的研究探讨不同的电话号码或扫描会话的运行，14，18，27，如何学习改变反馈范式如何影响学习28，是否一个给定的生物反馈治疗在脑功能的变化，坚持超越，最终协议结果引起学习生物反馈训练期间15，18，27，29然而，需要更大量的工作沿着这些路线是，在心中的最佳协议，根据对大脑的目标区域可能会有所不同，人口研究，以及其他变量。 神经反馈研究面临的挑战之一是最佳的方式来控制的实践中，曝光，动机和安慰剂效应。有各种各样的方法已在文献中描述的，每一种都有它的ADVAN产品关键词和缺点。在这个协议中，一个深水生物反馈范式，对照组收到作为其配套的生物反馈科目相同的刺激和领导相信他们接受自己的大脑活动模式为基础的真正的生物反馈。这种方法的优势，指示和刺激控制。它还有助于控制动机和安慰剂效应。也就是说，神经反馈诱使一个游戏的心态，在其成为个人投资在他们的表现在许多学科。假的控制条件下重复，尽可能密切的经验，从而为控制高层次的动机的神经反馈科目。此外，如果一个主体接收到的反馈表明越来越多的成功，随之而来的成就和自我控制的看法可以转化为行为的措施上的安慰剂效应。再次，假的范例，我们使用这种可能性为有效控制LY为可能。然而，这个深水生物反馈方法的缺点是，它积极误导科目，从而干扰与学习，通常会发生在实践期间无反馈。用于神经反馈研究的另一个控制条件是有科目无神经反馈执行相同的任务。这种实践和曝光效果的控制，并没有误导和可能混淆的自我反思的学习过程的缺点。但是，它可能无法控制以及动机和安慰剂效应。另一种形式的深水生物反馈也被用来在其中，虽然主题是误导，相信他们的目标区域是任务有关的科目就没有想过要参与任务的另一个大脑区域接收信息。这种方法涉及的任务无关的大脑区域的研究人员的假设，而这可能会产生问题，如果该地区涂RNS，参与任务。此外，如果该地区真的是无关紧要的任务，假科目是不可能成功控制，并因此可能感到失望和沮丧谁更有可能的成功经验，并感到满意的真实反馈科目，在控制。因此，这种形式的深水生物反馈不控制为主体的情绪状态（因此的动机和安慰剂效应）以及深水这个手稿中描述的类型，并有可能干扰学习过程中的相同缺点提供错误的信息。最后，控制条件，其中对照组接受治疗以外的磁铁的另一种形式（如认知行为疗法）可以用来对比的RT - fMRI的生物反馈，无论是治疗的黄金标准的成效目前。这最后的方法并不试图精确控制所有发生在神经反馈，从而影响不解决，无论是本身的反馈，诱导行为改善，而是要求的重要问题:采取一切在一起，可以实时功能磁共振成像生物反馈干预产生较好的临床或行为的结果比目前的替代品？总之，选择哪种类型的控制使用在生物反馈研究，是一个重要和具有挑战性的研究设计的方面，并解释结果时的控制条件的限制需要考虑。 虽然仍处于发育阶段，使用实时功能磁共振成像的生物反馈治疗一系列神经精神条件的潜在效用。此外，在评估脑组织的功能和认知/临床变量（生物反馈治疗前后收集）一起使用时，它可以是一个强有力的研究工具。从本质上讲，生物反馈亲志愿组织一个低风险的"扰动和测量"的方法来研究人的心理功能的神经基础:生物反馈是用来扰乱大脑的功能组织和，心理功能产生的变化进行测量。鉴于其研究和临床应用前景，这是一种很有前途的精神病学和认知神经科学领域的新技术。 该协议描述在这里调查的实时功能磁共振成像的生物反馈是否能帮助正常人的增益控制其污染焦虑。虽然它是可能的污染健康对照组的焦虑的神经基板是强迫症患者的不同，它也可能是强迫症状维度同时通过健康和病人的人群运行，并有类似的机制在这两个污染焦虑组。如果是这样，如果生物反馈的实时功能磁共振成像能有效地帮助正常人控制他们的焦虑，imilar范式可能有强迫症的临床效用。

real-time fmri biofeedback targeting the orbitofrontal cortex for contamination anxiety

我们目前的训练科目，以控制其使用生物反馈的实时功能磁共振成像（RT - fMRI）技术，数据焦虑与污染相关的前额皮质区域活动的一种方法。增加本地区的活动是可见的污染，无论是在对照组1和在个人与强迫症（OCD），2一种比较常见的，往往衰弱的精神障碍，涉及污染的焦虑焦虑的关系。虽然一直在强迫症，异常牵连许多大脑区域在前额皮层（OFC）是最一致的发现之一。3，4此外，在的OFC已发现多动症与强迫症的症状严重程度相关，并在多动症跌幅这一地区已报告相关症状严重程度下降，因此，能够控制这个大脑区域可能转化为CL强迫症状inical的改善，包括污染焦虑。 RT - fMRI数据，生物反馈是一种新技术，在活动的时空格局，在一个特定的地区，在主体的大脑（或分布的大脑活动模式与特定的关联）作为反馈信号提供的主体。最近的报告表明，人们能够控制在特定的大脑区域的活动发展，与RT - fMRI的生物反馈提供。7-12尤其是使用这种技术的目标参与情绪加工的脑区，一些研究报告在训练科目成功控制这些地区。13-18在一些情况下，RT - fMRI的生物反馈训练已报诱导受试者的认知，情感，或临床变化，8，9，13，19日在这里我们说明这种技术适用于治疗在健康受试者的污染焦虑。这种生物反馈干预将是一个宝贵的BASIC的研究工具:它允许扰乱脑功能的研究人员，测量大脑动态产生的变化，并与那些污染焦虑或其他行为的措施的变化。此外，这种方法的建立作为一个对作为一个强迫症的治疗干预的功能磁共振成像为基础的生物反馈调查的第一步。由于强迫症患者的四分之一，大约收到从目前可用的治疗，20日至22日的形式，没有什么好处，那些做的好处很少完全恢复，新方法，为治疗这种人口急需。

Watch this Scientific Journal Video about Real-time fMRI Biofeedback Targeting the Orbitofrontal Cortex for Contamination Anxiety at JoVE.com

Real-time fMRI Biofeedback Targeting the Orbitofrontal Cortex for Contamination Anxiety

在这里，我们提出了一个方法训练的人来控制污染焦虑的一个大脑区域和探测污染的焦虑和大脑连接模式之间的关系。

实时功能磁共振成像生物反馈，针对污染焦虑的眶额皮层

We present a method for training subjects to control activity in a region of their orbitofrontal cortex associated with contamination anxiety using ...

real-time-fmri-biofeedback-targeting-orbitofrontal-cortex-for

Nanyang Technological University

Research

JoVE Journal

Medicine

20.7K 次观看.  Yale University School of Medicine. 在这里，我们提出了一个方法训练的人来控制污染焦虑的一个大脑区域和探测污染的焦虑和大脑连接模式之间的关系。

视频: Real-time fMRI Biofeedback Targeting the Orbitofrontal Cortex for Contamination Anxiety

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

多光谱实时荧光成像术在妇科肿瘤前哨淋巴结的检测

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

科研

医学

Evaluation of Nanoparticle Uptake in Tumors in Real Time Using Intravital Imaging

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle uptake in tumors at the microscopic level1,2,3, highlighting the utility of a suitable xenograft model in which to perform detailed uptake analyses. Here, we use high-resolution intravital imaging to evaluate nanoparticle uptake in human tumor xenografts in a modified, shell-less chicken embryo model. The chicken embryo model is particularly well-suited for these in vivo analyses because it supports the growth of human tumors, is relatively inexpensive and does not require anesthetization or surgery 4,5. Tumor cells form fully vascularized xenografts within 7 days when implanted into the chorioallantoic membrane (CAM) 6. The resulting tumors are visualized by non-invasive real-time, high-resolution imaging that can be maintained for up to 72 hours with little impact on either the host or tumor systems. Nanoparticles with a wide range of sizes and formulations administered distal to the tumor can be visualized and quantified as they flow through the bloodstream, extravasate from leaky tumor vasculature, and accumulate at the tumor site. We describe here the analysis of nanoparticles derived from Cowpea mosaic virus (CPMV) decorated with near-infrared fluorescent dyes and/or polyethylene glycol polymers (PEG) 7, 8, 9,10,11. Upon intravenous administration, these viral nanoparticles are rapidly internalized by endothelial cells, resulting in global labeling of the vasculature both outside and within the tumor7,12. PEGylation of the viral nanoparticles increases their plasma half-life, extends their time in the circulation, and ultimately enhances their accumulation in tumors via the enhanced permeability and retention (EPR) effect 7, 10,11. The rate and extent of accumulation of nanoparticles in a tumor is measured over time using image analysis software. This technique provides a method to both visualize and quantify nanoparticle dynamics in human tumors.

We present a novel approach to quantify nanoparticle localization in the vasculature of human xenografted tumors using dynamic, real-time intravital imaging in an avian embryo model.

使用实时活体成像在肿瘤纳米粒子吸收的评价

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle ...

Remote Magnetic Navigation for Accurate, Real-time Catheter Positioning and Ablation in Cardiac Electrophysiology Procedures

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac substrates such as left atrial flutter. This protocol describes all the clinical and invasive interventional steps performed during a human electrophysiological study and ablation to assess the accuracy, safety and real-time navigation of the Catheter Guidance, Control and Imaging (CGCI) system. Patients who underwent ablation of a right or left atrium flutter substrate were included. Specifically, data from three left atrial flutter and two counterclockwise right atrial flutter procedures are shown in this report. One representative left atrial flutter procedure is shown in the movie. This system is based on eight coil-core electromagnets, which generate a dynamic magnetic field focused on the heart. Remote navigation by rapid changes (msec) in the magnetic field magnitude and a very flexible magnetized catheter allow real-time closed-loop integration and accurate, stable positioning and ablation of the arrhythmogenic substrate.

This report provides a detailed description of a new remote navigation system based on magnetic driven forces, which has been recently introduced as a new robotic tool for human cardiac electrophysiology procedures.

遥控磁导航导管定位准确，实时和心脏电生理消融程序

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac ...

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process including development1, apoptosis2, and cell cycle regulation3. MiRNAs are estimated to regulate the expression of 30% to 90% of human genes4 by binding to their target messenger RNAs (mRNAs)5. Widespread dysregulation of miRNAs has been reported in various diseases and cancer subtypes6. Due to their prevalence and unique structure, these small molecules are likely to be the next generation of biomarkers, therapeutic agents and/or targets.
Methods used to investigate miRNA expression include SYBR green I dye- based as well as Taqman-probe based qPCR. If miRNAs are to be effectively used in the clinical setting, it is imperative that their detection in fresh and/or archived clinical samples be accurate, reproducible, and specific. qPCR has been widely used for validating expression of miRNAs in whole genome analyses such as microarray studies7. The samples used in this protocol were from patients who underwent radical prostatectomy for clinically localized prostate cancer; however other tissues and cell lines can be substituted in. Prostate specimens were snap-frozen in liquid nitrogen after resection. Clinical variables and follow-up information for each patient were collected for subsequent analysis8.
Quantification of miRNA levels in prostate tumor samples. The main steps in qPCR analysis of tumors are: Total RNA extraction, cDNA synthesis, and detection of qPCR products using miRNA-specific primers. Total RNA, which includes mRNA, miRNA, and other small RNAs were extracted from specimens using TRIzol reagent. Qiagen's miScript System was used to synthesize cDNA and perform qPCR (Figure 1). Endogenous miRNAs are not polyadenylated, therefore during the reverse transcription process, a poly(A) polymerase polyadenylates the miRNA. The miRNA is used as a template to synthesize cDNA using oligo-dT and Reverse Transcriptase. A universal tag sequence on the 5' end of oligo-dT primers facilitates the amplification of cDNA in the PCR step. PCR product amplification is detected by the level of fluorescence emitted by SYBR Green, a dye which intercalates into double stranded DNA. Specific miRNA primers, along with a Universal Primer that binds to the universal tag sequence will amplify specific miRNA sequences. 
The miScript Primer Assays are available for over a thousand human-specific miRNAs, and hundreds of murine-specific miRNAs. Relative quantification method was used here to quantify the expression of miRNAs. To correct for variability amongst different samples, expression levels of a target miRNA is normalized to the expression levels of a reference gene. The choice of a gene on which to normalize the expression of targets is critical in relative quantification method of analysis. Examples of reference genes typically used in this capacity are the small RNAs RNU6B, RNU44, and RNU48 as they are considered to be stably expressed across most samples. In this protocol, RNU6B is used as the reference gene.

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR

Quantitative Real Time polymerase chain reaction (qPCR) is a rapid and sensitive method to investigate the expression levels of various microRNA (miRNA) molecules in tumor samples. Using this method expression of hundreds of different miRNA molecules can be amplified, quantified, and analyzed from the same cDNA template.

实时定量PCR（qPCR的microRNA检测在前列腺肿瘤）

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process ...

Movement Retraining using Real-time Feedback of Performance

Any modification of movement - especially movement patterns that have been honed over a number of years - requires re-organization of the neuromuscular patterns responsible for governing the movement performance. This motor learning can be enhanced through a number of methods that are utilized in research and clinical settings alike. In general, verbal feedback of performance in real-time or knowledge of results following movement is commonly used clinically as a preliminary means of instilling motor learning. Depending on patient preference and learning style, visual feedback (e.g. through use of a mirror or different types of video) or proprioceptive guidance utilizing therapist touch, are used to supplement verbal instructions from the therapist. Indeed, a combination of these forms of feedback is commonplace in the clinical setting to facilitate motor learning and optimize outcomes.
Laboratory-based, quantitative motion analysis has been a mainstay in research settings to provide accurate and objective analysis of a variety of movements in healthy and injured populations. While the actual mechanisms of capturing the movements may differ, all current motion analysis systems rely on the ability to track the movement of body segments and joints and to use established equations of motion to quantify key movement patterns. Due to limitations in acquisition and processing speed, analysis and description of the movements has traditionally occurred offline after completion of a given testing session.
This paper will highlight a new supplement to standard motion analysis techniques that relies on the near instantaneous assessment and quantification of movement patterns and the display of specific movement characteristics to the patient during a movement analysis session. As a result, this novel technique can provide a new method of feedback delivery that has advantages over currently used feedback methods.

Retraining abnormal movement patterns following injury or disease is a key component of physical rehabilitation. Recent advances in technology have permitted accurate assessment of movement during a variety of tasks, with near instantaneous quantification of results. This provides new opportunities for modification of faulty movement patterns in real time.

的运动再培训采用实时反馈的性能

Any modification of movement - especially movement patterns that have been honed over a number of years - requires re-organization of the neuromuscular ...

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Intravital imaging has emerged as an indispensable tool in biological research. In the process, many imaging techniques have been developed to study different biological processes in animals non-invasively. However, a major technical limitation in existing intravital imaging modalities is the inability to combine non-invasive, longitudinal imaging with single-cell resolution capabilities. We show here how transplantation into the anterior chamber of the eye circumvents such significant limitation offering a versatile experimental platform that enables non-invasive, longitudinal imaging with cellular resolution in vivo. We demonstrate the transplantation procedure in the mouse and provide representative results using a model with clinical relevance, namely pancreatic islet transplantation. In addition to enabling direct visualization in a variety of tissues transplanted into the anterior chamber of the eye, this approach provides a platform to screen drugs by performing long-term follow up and monitoring in target tissues. Because of its versatility, tissue/cell transplantation into the anterior chamber of the eye not only benefits transplantation therapies, it extends to other in vivo applications to study physiological and pathophysiological processes such as signal transduction and cancer or autoimmune disease development.

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

A new approach combining intraocular transplantation and confocal microscopy enables longitudinal, non-invasive real-time imaging with single-cell resolution within grafted tissues in vivo. We demonstrate how to transplant pancreatic islets into the anterior chamber of the mouse eye.

纵向，非侵入性的移植到眼睛的前房<em在体内</em&gt;单细胞分辨率的实时成像

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their translational repression. Changes in miRNA expression have been shown to be involved in the development of all major complex diseases. Furthermore, recent findings showed that miRNAs can be secreted to the extracellular environment and enter the bloodstream and other body fluids where they can circulate with high stability. The function of such circulating miRNAs remains largely elusive, but systematic high throughput approaches, such as miRNA profiling arrays, have lead to the identification of miRNA signatures in several pathological conditions, including neurodegenerative disorders and several types of cancers. In this context, the identification of miRNA expression profile in the cerebrospinal fluid, as reported in our recent study, makes miRNAs attractive candidates for biomarker analysis.
There are several tools available for profiling microRNAs, such as microarrays, quantitative real-time PCR (qPCR), and deep sequencing. Here, we describe a sensitive method to profile microRNAs in cerebrospinal fluids by quantitative real-time PCR. We used the Exiqon microRNA ready-to-use PCR human panels I and II V2.R, which allows detection of 742 unique human microRNAs. We performed the arrays in triplicate runs and we processed and analyzed data using the GenEx Professional 5 software.
Using this protocol, we have successfully profiled microRNAs in various types of cell lines and primary cells, CSF, plasma, and formalin-fixed paraffin-embedded tissues.

We describe a protocol of real time PCR to profile microRNAs in the cerebrospinal fluid (CSF). With the exception of RNA extraction protocols, the procedure can be extended to RNA extracted from other body fluids, cultured cells, or tissue specimens.

脑脊液的microRNA分析采用实时定量PCR技术

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

实时的细胞毒性分析在人全血

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.

This protocol describes the use of multiphoton microscopy to perform extended time-lapse imaging of multicellular interactions in real time, in vivo at single cell resolution.

实时多细胞动力学延长时间推移活体成像在肿瘤微环境

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...