NIH-NINDS K23格兰特NS044936（JMS），资助这项研究是由美国癫痫基金会，医学计算和综合生物医学研究中心（CIBR）种子奖助金（ZH）贝勒医学院提供的; ，并在莱夫家庭基金会（JMS） 数据采集提供了协助:伊丽莎白·皮尔斯（加州大学洛杉矶分校）。

1。主题<ol><li> 36个学科的研究对象包括三组:右颞叶癫痫（11例），左颞叶癫痫（12例）和健康对照组（n = 13）。所有受试者获得书面知情同意书。该研究遵循美国加州大学洛杉矶分校（UCLA）的机构审查委员会的指导方针。 </li><li>癫痫学科组应该是病人谁是候选人，前颞叶切除术通过视频脑电图监测，头颅MRI，PET成像和神经心理测试确定。患者应在功能磁共振成像扫描过程中继续他们一贯的药物，不应该立即扫描下面的癫痫发作。确保所有受试者有正常的大脑核磁共振成像，且无神经系统疾病（癫痫比在患者组除外）或正在使用神经系统药物。 </li></ol> 2。成像<ol><li>使用3特斯拉MRI系统的成像。获得轴向切片图像功能使用平面回波成像荷兰国际集团（EPI）序列，并使用一个被宠坏的梯度解剖影像回忆（SPGR）序列。 </li><li>使用以下参数进行功能成像:TR = 2000毫秒，TE = 30毫秒，FOV = 210毫米，矩阵= 64×64，切片厚度为4毫米，34片。用于高分辨率成像的结构参数如下:TR = 20毫秒，TE = 3毫秒，FOV = 256毫米，矩阵= 256×256，切片厚度为1毫米，160片。 </li><li>每个成像会议应持续20分钟。请参与者放松闭眼。没有特殊的听觉输入是必需的。 </li></ol> BOLD数据3。预处理<ol><li>预处理采用FSL（fMRIB软件库）软件版本4.1.6（英国牛津，www.fmrib.ox.ac.uk / FSL）13,14 fMRI数据。预处理步骤应包括以下内容:使用FSL MCFLIRT去除头部运动伪影15。使用FSL BET除去非脑组织16与BET选项-F大胆的文件。这有助于一个中运行F单单脑组织urther分析步骤。 </li><li>在壮举，运行，注册一个微加工分析。选择"第一层次的分析"，改变"全分析"，从顶部的两个按钮"预统计"。 <ol><li>根据预先统计选项卡，取消选中"下注脑提取"，然后选择"无"为"动态校正"（因为这些都已经做了以上）。注册功能（BOLD）的图像的解剖（SPGR）的图像，然后以一个标准（MNI）图像。这将导致在变换矩阵，它是在分析过程中用于以后翘曲在标准空间选入受试者脑空间的种子的产生。 </li></ol></li><li>使用生成的变换矩阵（名为"standard2example_func.mat"）和转换CSF和白质的ROI为单独的BOLD空间。 <ol><li>从使用fslmeants命令CSF和白质的ROI提取的时间序列，使用投资回报率在个人主体空间作为掩膜。使用标准化的软件"R"提取的时间序列。这些时间序列被用作在GLM回归后，从分析中删除对应的伪迹信号。 </li></ol></li><li>接下来的步骤是去除受运动有关的文物。对于运动参数的回归，设置运行它之前在佛山照明FEAT以下。 <ol><li>在数据选项卡中，使用运动校正和脑提取文件作为输入，设置TR值，以符合您的数据集。设置高通使用的是100秒过滤器过滤。高通滤波将删除不感兴趣的信号，这是非常低的频率。一个低通滤波器，以滤除高频信号将在步骤4.1以后可以应用。 </li><li>在预统计选项卡中，选择"无"下的"修正议案"，因为它已经完成。取消选中"下注脑提取"，因为它已经完成。进行空间平滑使用5毫米全宽半高（FWHM）。 </li><li>内的统计选项卡，倒退6运动参数和它们的时间导数。选择"无"为卷积，并检查"应用时间滤波。"使用FSL MCFLIRT的输出来获得运动参数的文本文件，它可以是那么投入的壮举分析模型，以在一般线性模型（GLM）这些回归</li><li>此外，还添加了提取，并在标准化之前的步骤，就GLM脑脊液和白质信号。选择"无"为卷积，加上时间导数，并取消选中"应用时间滤波"。 </li></ol></li></ol> 4，统计方法<ol><li>从上面所描述的预处理的残差应该用于基于种子的相关性。这些残差应通过0.1赫兹的低通滤波器被首先通过，然后通过减去平均值，通过标准偏差除以，然后通过添加缩放贬低100。种子应的直径为6毫米用MRICron软件的标准MNI空间来限定。 </li><li>在后部和前部的种子应该对应于以下坐标:（1）Rsp时/ PCUN区域（x = 2时，Y = -60，Z = 36）和（2）腹内侧前额叶皮质（VMPFC，X = 3，Y = 60，Z = -1）。这些种子位置已被定义健康对照组中，并转化到主体空间中的下一个步骤17-19。 <ol><li>种子应随后从标准MNI空间转化为每个主题的个别功能的大脑空间。为此，请使用上面生成的变换矩阵（名为"standard2example_func.mat"）从标准（MNI）空间变换种子的个别功能（BOLD）的空间。 </li><li>使用fslmeants命令从先前贬低和缩放残留，使用的种子在个人主体空间作为掩膜提取的时间序列。使用软标准化提取的时间序列洁具"R"。 </li></ol></li><li>种子体素和所有其他的脑体素之间的偏相关系数应分别为每个主题对每个运行计算。对于这一点，FSL FEAT GUI中，选择"第一层次的分析"，然后在"统计+后统计"。在数据选项卡，以前贬低和缩放残留应作为投入的壮举。 </li><li>设置高通滤波器的截止到10,000时，如残留的已经高的通过在100秒。在状态标签，取消勾选"使用胶片白化"，并使用以前提取，归一化种子时间序列中的GLM。内后统计选项卡中，设置所需的Z-统计阈值的2.0的值。 </li><li>在此之前运行组分析相结合的学科内运行，一Fisher的Z变换应该从先前运行的相关分析（步骤4.3）中产生的COPE（参数估计值的对比）文件执行。复制登记资料FROM的壮举分析的"REG"目录中的步骤做3.1到4.3级的相关运行。 </li><li>通过结合各学科内运行运行一个更高层次的分析。对于这一点，FSL FEAT GUI中，选择"更高层次的分析"，然后在"统计+后统计"。在"数据"选项卡上，选择"输入是低层次FEAT目录"，并从步骤4.4输入对象的运行。内的"属性"选项卡中，选择"混合效应:简单OLS"。建立模型，均值效果;每个对象的运行中输入值1。 </li><li>结合以上数据的运行主体，普通最小二乘法（OLS）之间的简单混合效应分析应该被使用。对于这一点，FSL FEAT GUI中，选择"高层次的分析"和"统计+后统计"。在数据选项卡上，选择"输入是低层次FEAT目录"，进入受试者的步骤4.5结合运行​​。 </li><li>在状态标签，选择"混合效果:简单OLS"&#39;建立一个模型，3组;每个科目所属的组，否则为0输入1的值。组分析应于每个像素使用一个单向ANOVA与三级这对应于三个组（TLE右，左颞叶癫痫，与健康对照）来完成。 </li><li>阈值与Z统计量的图像使用集群形成z&gt; 2.0的阈值和修正的p集群显著门槛= 0.05 20。要获得相关地图上正确的z值，逆Fisher的Z变换应该对结果进行。 </li><li>下面具体对比应该比较（1）右边TLE&gt;控制; （2）左侧颞叶癫痫&gt;控制; （3）右​​颞叶癫痫&gt;左颞叶癫痫; （4）左颞叶癫痫&gt;右颞叶癫痫; （5）控制&gt;右颞叶癫痫; （6）控制&gt;左颞叶癫痫; （7）TLE（合并左，右）&gt;控制;和（7）的控制&gt; TLE（合并右和左）。 </li></ol>

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Brain Mapp. 32, 883-895 (2011).</li><li>Pereira, F. R., 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=Asymmetrical+hippocampal+connectivity+in+mesial+temporal+lobe+epilepsy:+evidence+from+resting+state+fMRI.">Asymmetrical hippocampal connectivity in mesial temporal lobe epilepsy: evidence from resting state fMRI.</a> BMC Neurosci. 11, 1-13 (2010).</li><li>Dupont, S., 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=Bilateral+hemispheric+alteration+of+memory+processes+in+right+medial+temporal+lobe+epilepsy.">Bilateral hemispheric alteration of memory processes in right medial temporal lobe epilepsy.</a> J. Neurol. Neurosurg. Psychiatr. 73, 478-485 (2002).</li><li>Vlooswijk, M. 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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Default+mode+network+connectivity+indicates+episodic+memory+capacity+in+mesial+temporal+lobe+epilepsy.">Default mode network connectivity indicates episodic memory capacity in mesial temporal lobe epilepsy.</a> Epilepsia. 54, (2013).</li><li>Zhang, Z., 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=Altered+spontaneous+neuronal+activity+of+the+default-mode+network+in+mesial+temporal+lobe+epilepsy.">Altered spontaneous neuronal activity of the default-mode network in mesial temporal lobe epilepsy.</a> Brain Res. 1323, 152-160 (2010).</li><li>Horovitz, S. G., 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=Decoupling+of+the+brain's+default+mode+network+during+deep+sleep.">Decoupling of the brain's default mode network during deep sleep.</a> Proc. Natl. Acad. Sci. U.S.A. 106, 11376-11381 (2009).</li><li>Deshpande, G., Kerssens, C., Sebel, P. S., Hu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+local+coherence+in+the+default+mode+network+due+to+sevoflurane+anesthesia.">Altered local coherence in the default mode network due to sevoflurane anesthesia.</a> Brain Res. 1318, 110-121 (2010).</li></ol>

恩格尔博士是由美国国立卫生研究院资助P01 NS02808，R01 NS33310和NS42372 U01资助，拥有专利WO 2009/123734A1和WO 2009/123735A1，接收来自MedLink，威科，布莱克威尔，以及爱思唯尔特许权使用费，并已收到酬金从Medtronics ，威科，和最好的医生。斯特恩博士曾担任高薪顾问，UCB和Lundbeck公司。斯特恩博士是MedLink神经学的编辑，并已获得特许权使用费从威科和麦格劳 - 希尔。其余的作者没有披露或利益冲突申报。

癫痫被认为是一个网络的疾病，以及涉及网络的异常是存在癫痫发作过程中，并在间期状态21。基于任务的功能磁共振成像已被用来分析在TLE 8的语言和存储网络的异常。 FcMRI有着先天的优势，在学习DMN 12，因为它是一种网络主要活跃在静止状态。该DMN是大脑区域已经被认为是活跃在谁是静置并从事自发的思绪清醒的个人网络。这些区域被示出为包括在VMPFC，Rsp时/ PCUN，后扣带回皮质，顶叶区域，侧颞区，海马12,17。的DMN被认为是衬底的昏迷状态。已经报道在影响意识，记忆和社会认知如失神发作，阿尔茨海默氏病，和孤独症/精神分裂症，respecti条件被改变vely 22-26。 颞叶是错综复杂的连接到DMN。该DMN的二次介入可能介导颞叶癫痫对认知和意识12,27的影响。的DMN的这种次要的参与也可以是致病对于其他条件影响颞叶/边缘系统，例如精神分裂症25和阿尔茨海默氏病22,26的临床效果。此前的研究表明DMN参与颞叶癫痫和其他形式的癫痫发作过程中，在发作间期癫痫样放电，并在间期状态9,28-33。这些研究表明DMN在间期脑电图及颞叶癫痫的发作行为特征潜在的重要作用。在目前的实验中，基于种子的fcMRI是用来分析的DMN在TLE中的基础状态，以评估TLE对DMN的效果。该DMN的枢纽和辐射模型假定，它的主要子系统后（RSP / PCUN）和前（VMPFC）组件。在这里，这两个主要区域的连通性特征逐一检查，以评估TLE如何影响他们的互联互通和它们相关的子网。 当前的基于种子的分析揭示了DMN的前部和后部轮毂之间的连接断开。这种功能连接减少很可能与在所涉及的结构之间的解剖白质连接性的减少作为已被证明在一项研究显示同时减少功能和白质连接后DMN和患者的颞叶癫痫34海马之间。此外，它表明一般在DMN到其他脑区的前部和后部轮毂的左TLE增加的连接和通常在右颞叶癫痫的连通减小。这与以前的研究显示在右侧TLE减少连通性和代偿性增大相一致的连接在左颞叶癫痫30,35。一般来说，右颞叶癫痫往往涉及双边结构，而左颞叶癫痫涉及功能活化36,37的再分配。这一点尤其突出在前种子在那里留下TLE具有更大的连接围鳃盖范畴方面，对比右侧颞叶癫痫（ 图1G，1I和1J）。此，结合后的种子在左TLE下部连通与对照组相比（ 图1F和1K），表明在左侧TLE增加前的连接，可能会导致从后种子的断开或后部和前部之间的连接种子。海马是通过后路DMN 34连接到前部DMN。 在本研究中所使用的技术可以通过改变种子位置并产生对应于其他脑区大脑网络进行修改。人ternative分析软件，可以用来做类似的分析（ 例如 ，SPM，AFNI）。种基于相关性分析，它们需要有关的假定网络在调查了事先确定的种子位置的结构的假说是有限的。如果基础的假设是有缺陷的，结果不会是进口的。 静息状态fcMRI 2可以在BOLD信号提供功能连接的测量基础上的自发调制（ 即t向无关）。两个用于此目的的常用方法是种子（感兴趣区域）的相关性和ICA 6。基于种子相关分析需要关于推定的网络的被调查了事先确定的种子位置的结构的假说。选择的种子位置和提取构成所述种子，整个大脑的一个体素逐体素分析的体素的BOLD信号后被执行，以确定其他脑区相似的BOLD信号的图案，并且这阐明以相互关联的网络。种子分析的另一种方法是由两个或多个感兴趣，区域之间的相关联的信号。使用这样的方法，该连接被发现的病变海马和在TLE 38后DMN之间有所减少。另一方面，ICA是数据没有预先存在的假设驱动的并描述空间 - 时间不同的网络的静息状态大脑内。也被使用ICA进行DMN在TLE的研究，并确定患者右前方DMN的降低连接和左颞叶癫痫与对照组相比。然而，我们的投资回报率（种子）为基础的分析显示前DMN网络的TLE的存在，这实际上是更广泛的在左颞叶癫痫与对照组相比。这种差异可以从后部DMN，其中一个分离的前部NETW有关前的建议断开进行评估时，主要由后DMN连接识别网络ORK是不太明显的。与先前的研究一致，我们可以复制在TLE 39颞参与。 当前分析表明基于种子的fcMRI如何可以用来探测大脑网络中脑障碍如TLE。这类基团的不同的识别有助于理解功能异常的疾病状态打开的该实施的分析，可以在单一主体级别2被解释未来应用的可能性。 这种分析的一个限制是无法确定被摄物体的清醒/睡眠状态。睡眠已经与额叶皮质与深睡40或全身麻醉41的降低DMN连接相关联。进一步的研究是必要的，以探讨是否这些发现可以通过控制睡眠状态被复制。

功能性磁共振成像连接（fcMRI）是一个相对较新的分析方法，量化的基础上他们的血氧水平依赖（BOLD）的信号时间序列的相似性不同大脑区域之间的关系fMRI数据 - 这就是所谓的"功能性"的连接，并且是从解剖学的连接，它描述的区域（ 例如 ，白质纤维）之间的物理连接的存在区分开来。在这种方法中的一种特殊应用，时间序列收集时，参与者不从事一项任务或在所谓的"休息状态"。 虽然在1995年1首次描述，出现了fcMRI导致有关2012年该技术大约1,000出版物兴趣盎然。fcMRI拥有基于任务的功能磁共振成像的内在利益（1）没有要执行特定的任务，（ 2）待合作没有必要的，（3）的数据集可以被用来查询几个不同的网络，（4）较好的信噪比存在可能是由于在所涉及的脑能量学的差异，和（5）的规避与任务相关的困惑2。作为一个概念证明，fcMRI改变已被证明与脑电图3和局部场电位在大脑的变化4对应。 fcMRI分析技术包括投资回报率/种子为基础的技术，独立成分分析（ICA），图论分析，Granger因果关系分析，本地方法（低频波动，区域同质化分析的幅度），及其他5。没有任何单一的技术尚未表现出明显优于另一种，但最常用的方法是基于种子和ICA方法6。基于种子的fcMRI从相关研究中的假定网络的预选部分BOLD信号的时间波动称为"种子1;或"感兴趣区域（ROI）"到大脑的其他部分。大脑呈现BOLD信号关联到种子区域的区域被认为是划分所涉及的网络部分。相比之下，ICA使用无模型数据驱动分析通过分析全脑5的血流动力学信号特征提取空间-时间相关的大脑区域（独立组件，集成电路）。 在目前的手稿，在颞叶癫痫中的DMN的静息状态的种子为基础的连通性分析的先前发表的研究使用的方法的描述，提出7。 TLE是成人癫痫的最常见形式。除了 ​​癫痫发作，颞叶癫痫导致多发性脑网络，包括记忆，行为，思想和感觉功能8功能障碍。该DMN由subserving意识，静息状态的认知脑区构成。的DMN已报道涉及具有降低consc关联性发作iousness 9,10。此外，海马是参与TLE该键结构和已被认为是DMN的成分。然而，PCC向海马结构的连通性比与其它DMN的成分，如内侧前额叶和顶叶皮质弱。这表明，海马是DMN的任一个子网或一个相互作用网络11,12。颞叶癫痫和DMN之间的这些共性提出了DMN功能连接被改变在颞叶癫痫的可能性。这种分析的受试者TLE的DMN比较健康对照组，以深入了解DMN在TLE的参与。放置在DMN的主要集线器种子的连接-前部和后部轮毂的区域进行分析12。种子被放置在后部毂组成的retrosplenium /楔前叶（RSP / PCUN）以及前部毂组成的腹内侧前额叶皮质（VMPFC）在具有TLE患者和在健康对照来识别DMN的后部和前部的子网。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
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			<td>MRI machine</td>
		</tr>
		<tr>
			<td>Linux workstation with image analysis software installed</td>
		</tr>
	</tbody>
</table>

network analysis of the default mode network using functional connectivity mri in temporal lobe epilepsy

功能性磁共振成像的连接（fcMRI）是一个功能磁共振成像方法，它研究的基础上BOLD信号波动随时间的相关性不同的大脑区域的连通性。颞叶癫痫（TLE）是成人癫痫最常见的类型和涉及多个脑网络。默认模式网络（DMN）参与意识，静息状态下的认知和被认为是受影响颞叶癫痫发作的地方引起意识障碍。使用基于fcMRI种子中癫痫的DMN进行了检查。的DMN的前部和后部轮毂被用作种子在此分析。结果表明在基础状态的前部和颞叶癫痫中的DMN后路集线器之间的连接断开。此外，增加的DMN连接在左侧颞叶癫痫等脑区与右颞叶癫痫下降连通显露。分析表明基于种子的fcMRI如何可以用来探测大脑网络中脑障碍如TLE。

图1显示了DMN显示与连接从后部种子（RSP / PCUN，红-黄色）和前籽（VMPFC，蓝绿色的颜色），并在不同的题目组中发现的网络中进行比较（ 图1A-C）的以及彼此之间，即健康对照相比，所有患者的颞叶癫痫（ 图1D和1E），然后健康对照组分别相比，左颞叶癫痫（ 图1F和1G）和右颞叶癫痫（ 图1H和1I）。左，右颞叶癫痫之间的直接比较也显示（ 图1J及1K）。 TLE TLE包括具有左或右颞叶癫痫科目。与对照组相比，这种联合用药组已经从后减少连接到前DMN区域（ 图1D，红色）一个以及减少由前DMN区域连接到后部元件（ 图1D，蓝三色）。与TLE受试者也显示增加额叶-顶叶的连接与前部和后部DMN（ 图1E，蓝/红色）。 左侧颞叶癫痫 左颞叶癫痫主题已经降低后DMN的连接与前DMN区域和海马，海马旁回，脑干，和内侧枕叶皮层（ 图1F，红三种颜色）。使用前的种子，用后的组件（海马，海马旁回，梭状回，舌回，扣带回）减少连接也被看作（ 图1F，蓝三种颜色）。左颞叶癫痫受试者被发现有连接的前后种子（ 图1G，红色和蓝色的颜色）COMP扩大围鳃盖网ARED健康对照组。 右颞叶癫痫 如在受试者的左颞叶癫痫的情况下，那些具有合适的TLE减少了后部DMN种子的连接与前DMN组件（ 图1H，红色）。前种子减少了连接到后部区域（双侧海马，壳核，尾状核）和前DMN本身。 （ 图1H，蓝三种颜色）。用的前后种子在右颞叶癫痫提高了连接领域包括左外侧颞叶皮质，楔前叶，扣带和辅助运动皮层。 （ 图1I）。 右与左颞叶癫痫 右颞叶癫痫伴左颞叶癫痫直接比较发现后DMN种子提高了连接在左颞叶癫痫（ 图1J，红三种颜色），左缘上回以及前DMN种子额叶区（ 图1J，蓝三种颜色）相比，右侧颞叶癫痫。在右颞叶癫痫后种子提高了连接的领域包括双侧丘脑和脑干左侧海马，纺锤形和地区。前种子在右颞叶癫痫提高了连接的领域包括楔前叶，双侧丘脑区，脑干区域。 （ 图1K）。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51442/51442fig1highres.jpg" width="500" /> 图1。DMN的TLE比较健康对照组相比，使用后的种子（后DMN的Rsp时/ PCUN，红-黄色）和前籽（VMPFC，蓝绿色）的DMN连接显示在不同学科群（ 交流 ），TLE相比健康对照组（DE），左颞叶癫痫相比健康对照组（FG），右颞叶癫痫相比健康对照组（HI），右颞叶癫痫相比，左侧颞叶癫痫（JK）。 C-控制; L-左颞叶癫痫; R-右颞叶癫痫。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/51442/51442fig1highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy at JoVE.com

Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy

在默认模式网络（DMN）在颞叶癫痫（TLE）是在大脑的静息状态下使用基于种子的功能连通性磁共振成像（fcMRI）进行分析。

使用功能性磁共振成像连接在颞叶癫痫默认模式网络的网络分析

Functional connectivity MRI (fcMRI) is an fMRI method that examines the connectivity of different brain areas based on the correlation of BOLD signal ...

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17.8K 次观看.  Baylor College of Medicine. 在默认模式网络（DMN）在颞叶癫痫（TLE）是在大脑的静息状态下使用基于种子的功能连通性磁共振成像（fcMRI）进行分析。 

视频: Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.

Human tumor xenografts in immunodeficient mice are valuable tools to study cancer biology. Specific protocols to generate subcutaneous and intrahepatic xenografts from human hepatocellular carcinoma cells or tumor fragments are described. Liver regeneration induced by partial hepatectomy in recipient mice is presented as a strategy to facilitate intrahepatic engraftment.

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

科研

医学

Developing Neuroimaging Phenotypes of the Default Mode Network in PTSD: Integrating the Resting State, Working Memory, and Structural Connectivity

Complementary structural and functional neuroimaging techniques used to examine the Default Mode Network (DMN) could potentially improve assessments of psychiatric illness severity and provide added validity to the clinical diagnostic process. Recent neuroimaging research suggests that DMN processes may be disrupted in a number of stress-related psychiatric illnesses, such as posttraumatic stress disorder (PTSD).
Although specific DMN functions remain under investigation, it is generally thought to be involved in introspection and self-processing. In healthy individuals it exhibits greatest activity during periods of rest, with less activity, observed as deactivation, during cognitive tasks, e.g., working memory. This network consists of the medial prefrontal cortex, posterior cingulate cortex/precuneus, lateral parietal cortices and medial temporal regions.
Multiple functional and structural imaging approaches have been developed to study the DMN. These have unprecedented potential to further the understanding of the function and dysfunction of this network. Functional approaches, such as the evaluation of resting state connectivity and task-induced deactivation, have excellent potential to identify targeted neurocognitive and neuroaffective (functional) diagnostic markers and may indicate illness severity and prognosis with increased accuracy or specificity. Structural approaches, such as evaluation of morphometry and connectivity, may provide unique markers of etiology and long-term outcomes. Combined, functional and structural methods provide strong multimodal, complementary and synergistic approaches to develop valid DMN-based imaging phenotypes in stress-related psychiatric conditions. This protocol aims to integrate these methods to investigate DMN structure and function in PTSD, relating findings to illness severity and relevant clinical factors.

This protocol describes the complementary neuroimaging techniques of resting state structural connectivity, task-induced deactivation, and structural connectivity analyses to examine the default network in post-traumatic stress disorder. The use of synergistic methods could potentially lead to improved diagnostics and assessments of severity, outcome, and other relevant clinical factors.

发展中的默认模式网络的神经影像学表型在创伤后应激障碍:整合静息状态下，工作记忆和结构连接

Complementary structural and functional neuroimaging techniques used to examine the Default Mode Network (DMN) could potentially improve assessments of ...

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of women are alive 5 years after diagnosis. The omentum is a preferred site of HGSC metastasis formation. Despite the clinical importance of this microenvironment, the contribution of omental adipose tissue to ovarian cancer progression remains understudied. Omental adipose is unusual in that it contains structures known as milky spots, which are comprised of B, T, and NK cells, macrophages, and progenitor cells surrounding dense nests of vasculature. Milky spots play a key role in the physiologic functions of the omentum, which are required for peritoneal homeostasis. We have shown that milky spots also promote ovarian cancer metastatic colonization of peritoneal adipose, a key step in the development of peritoneal metastases. Here we describe the approaches we developed to evaluate and quantify milky spots in peritoneal adipose and study their functional contribution to ovarian cancer cell metastatic colonization of omental tissues both in vivo and ex vivo. These approaches are generalizable to additional mouse models and cell lines, thus enabling the study of ovarian cancer metastasis formation from initial localization of cells to milky spot structures to the development of widespread peritoneal metastases.

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

We outline a protocol that implements both in vivo and ex vivo approaches to study ovarian cancer colonization of peritoneal adipose tissues, particularly the omentum. Furthermore, we present a protocol to quantitate and analyze immune cell-structures in the omentum known as milky spots, which promote metastases of peritoneal adipose.

在体内和体外途径研究银河系结构现货卵巢癌转移定植腹膜脂肪

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of ...

MRI-guided dmPFC-rTMS as a Treatment for Treatment-resistant Major Depressive Disorder

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial prefrontal cortex (dmPFC) in patients with major depressive disorder (MDD). Technicians used a neuronavigation system to process patient MRIs to generate a 3-dimensional head model. The head model was subsequently used to identify patient-specific stimulatory targets. The dmPFC was stimulated daily for 20 sessions. Stimulation intensity was titrated to address scalp pain associated with rTMS. Weekly assessments were conducted on the patients using the Hamilton Rating Scale for Depression (HamD17) and Beck Depression Index II (BDI-II). Treatment-resistant MDD patients achieved significant improvements on both HAMD and BDI-II. Of note, angled, double-cone coil rTMS at 120% resting motor threshold allows for optimal stimulation of deeper midline prefrontal regions, which results in a possible therapeutic application for MDD. One major limitation of the rTMS field is the heterogeneity of treatment parameters across studies, including duty cycle, number of pulses per session and intensity. Further work should be done to clarify the effect of stimulation parameters on outcome. Future dmPFC-rTMS work should include sham-controlled studies to confirm its clinical efficacy in MDD.

Here we outline the procedure for MRI-guided repetitive transcranial magnetic stimulation to the dorsomedial prefrontal cortex as an experimental treatment for major depressive disorder.

MRI引导dmPFC-rTMS治疗作为治疗难治性抑郁症

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

在耐药性颞叶癫痫患者卵圆孔电极记录的网络分析

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Solubility of Hydrophobic Compounds in Aqueous Solution Using Combinations of Self-assembling Peptide and Amino Acid

Self-assembling peptides (SAPs) are promising vehicles for the delivery of hydrophobic therapeutics for clinical applications; their amphipathic properties allow them to dissolve hydrophobic compounds in the aqueous environment of the human body. However, self-assembling peptide solutions have poor blood compatibility (e.g., low osmolarity), hindering their clinical application through intravenous administrations. We have recently developed a generalized platform for hydrophobic drug delivery, which combines SAPs with amino acid solutions (SAP-AA) to enhance drug solubility and increase formulation osmolarity to reach the requirements for clinical uses. This formulation strategy was thoroughly tested in the context of three structurally different hydrophobic compounds &#8211; PP2, rottlerin, and curcumin &#8211; in order to demonstrate its versatility. Furthermore, we examined effects of changing formulation components by analyzing 6 different SAPs, 20 naturally existing amino acids at low and high concentrations, and two different co-solvents dimethyl sulfoxide (DMSO) and ethanol. Our strategy proved to be effective in optimizing components for a given hydrophobic drug, and therapeutic function of the formulated inhibitor, PP2, was observed both in vitro and in vivo. This manuscript outlines our generalized formulation method using SAP-AA combinations for hydrophobic compounds, and analysis of solubility as a first step towards potential use of these formulations in more functional studies. We include representative solubility results for formulation of the hydrophobic compound, curcumin, and discuss how our methodology serves as a platform for future biological studies and disease models.

This protocol describes a clinically-applicable means of dissolving hydrophobic compounds in an aqueous environment using combinations of self-assembling peptide and amino acid solutions. Our method resolves a major limitation of hydrophobic therapeutics, which lack safe, efficient means of solubility and delivery methods into clinical settings.

自组装肽与氨基酸结合的水溶液中疏水性化合物的溶解度

Self-assembling peptides (SAPs) are promising vehicles for the delivery of hydrophobic therapeutics for clinical applications; their amphipathic ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

小鼠颞叶癫痫的 Radiotelemetry 模型及脑电图监测

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&#8212;Application in Premanifest Huntington's Disease

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy begins many years prior to the motor onset (during the &#34;premanifest&#34; period), but the spatiotemporal pattern of regional atrophy across the brain has not been fully characterized. Here we demonstrate an online cloud-computing platform, &#34;MRICloud&#34;, which provides atlas-based whole-brain segmentation of T1-weighted images at multiple granularity levels, and thereby, enables us to access the regional features of brain anatomy. We then describe a regression model that detects statistically significant inflection points, at which regional brain atrophy starts to be noticeable, i.e. the &#34;change-point&#34;, with respect to a disease progression index. We used the CAG-age product (CAP) score to index the disease progression in HD patients. Change-point analysis of the volumetric measurements from the segmentation pipeline, therefore, provides important information of the order and pattern of structural atrophy across the brain. The paper illustrates the use of these techniques on T1-weighted MRI data of premanifest HD subjects from a large multicenter PREDICT-HD study. This design potentially has wide applications in a range of neurodegenerative diseases to investigate the dynamic changes of brain anatomy.

This paper describes a statistical model for volumetric MRI data analysis, which identifies the &#34;change-point&#34; when brain atrophy begins in premanifest Huntington&#39;s disease. Whole-brain mapping of the change-points is achieved based on brain volumes obtained using an atlas-based segmentation pipeline of T1-weighted images.

解剖性脑 MRI 的全脑分割及变化点分析--Premanifest 亨廷顿病的应用

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy ...

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further diagnose and manage PH, patients undergo repeated right heart catheterizations (RHC) wherein a Swan-Ganz catheter is advanced into a branch of the pulmonary artery and a balloon is inflated to wedge the catheter tip. This article illustrates a protocol whereby pulmonary artery endothelial cells (PAECs) may be harvested from the balloon tips of Swan-Ganz catheters after RHC, and purified with an anti- CD146 affinity column technique to purify putative PAECs. These cells might be used to provide an in situ snapshot of the biological state of the pulmonary vasculature endothelium to complement hemodynamic measurements obtained during RHC. Harvested and purified PAECs may be used for either cell culture or for subsequent analytical assays such as flow cytometery.

This protocol describes the removal and purification of pulmonary artery endothelial cells (PAECs) that adhere to the balloon tips of Swan-Ganz catheters while they are wedged in the pulmonary artery during right heart catheterization and remain adherent to the deflated balloon following removal from the patient's body.

右心导管插入术后天鹅管囊小贴中内皮细胞的提取

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further ...

Assessment of Static Graviceptive Perception in the Roll-Plane using the Subjective Visual Vertical Paradigm

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow the examination of all semicircular canals in the clinical setting. Assessment methods of the otolithic system, which is responsible for the perception of linear acceleration and perception of gravity, are far less in clinical use. There are several experimental approaches for measuring the perception of gravity. The most frequently used method is the determination of the subjective visual vertical. This is usually measured with the head in an upright position. We present here an assessment method for testing otolith function in the roll plane. The subjective visual vertical is measured in the head upright position as well as with head inclination of &#177; 15&#176; and &#177; 30&#176; in the roll plane. This extended functional paradigm is an easy-to-perform clinical test of otolith function and ensures increased information content for the detection of impaired graviceptive perception.

The perception of gravity is commonly determined by the subjective visual vertical in the head upright position. The additional assessment at head tilts of &#177; 15&#176; and &#177; 30&#176; in the roll plane ensures increased information content for the detection of impaired graviceptive perception.

使用主观视觉垂直范式对卷平面中的静态视觉感知的评估

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow ...