我们感谢强生约翰诺森律师事务所的顾问闫俊明，他负责研究资助。这项工作由吉林省科学技术厅（20220402076GH）资助。

该研究程序严格遵守吉林大学中日协和医院人体研究伦理委员会的规定。在Carto系统（3D映射系统）上搜索接受心房颤动射频消融的患者。然后，使用PACS系统确定患者在手术前是否进行了左心房CT检查，以确定每个选定的患者是否进行了左心房CT图像进行比较。Soundstar 是本研究中使用的 ICE 导管，3D 映射系统中提供了 Cartosound 模块。每位患者在 ICE 3D 建模前都提供了书面知情同意书。
1. 扫描前的准备
<ol><li>检查患者信息，例如准确的心率、节律、血压和血氧饱和度。让患者仰卧，双臂在两侧，大腿略微外展。</li>
	<li>在整个手术过程中，为所有患者提供芬太尼 （200 μg/mL） 的深度清醒镇静。选择右侧股静脉作为穿刺点，用2%利多卡因消毒并铺路进行局部麻醉。</li>
	<li>设置 ICE 导管：将 ICE 导管尾线与 3D 映射系统和超声波机器连接。在 3D 映射系统中打开“研究”界面，然后在“连接的机器”中选择“P500”。</li>
	<li>将ICE导管（直径：10f;超声模式：B模式）置于股静脉内。在推压过程中，实时监测超声波，确保导管处于安全空间。</li>
	<li>将消融导管传递到右心房，以形成右心房和冠状窦模型。将冠状动脉窦电极与 ICE 引导相结合放置冠状窦电极。</li>
	<li>ICE导管进入右心房后，确保左心房和左心耳结构在3D映射系统显示屏上显示左心房和左心耳结构，同时排除左心房血栓。</li>
	<li>插入房间隔穿刺针后，在ICE引导下确定最佳穿刺部位。然后，进行经鼻中隔穿刺。<ol><li>使用 “主页 ”视图确认 ICE 已到达右心房。然后软件显示左侧 PV 部分。单击 右弯 以显示下腔静脉和房间隔。</li>
		<li>将穿刺针鞘移至指向四点钟方向。在监测 ICE 的同时将针鞘缩回椭圆形窝，可以看到“帐篷征”。</li>
		<li>调整 ICE 导管以显示左侧 PV 为“兔耳征”。在ICE引导下，将针鞘慢慢撤回到椭圆形窝的下缘。</li>
		<li>顺时针旋转穿刺针以突破椭圆形窝。然后，通过房间隔穿刺针注射肝素生理盐水。 注意：在 LA 中观察到盐水泡，表明房间隔成功穿刺。</li>
	</ol></li>
</ol>左心房和肺静脉的2. 3D建模
注：ICE在两个方向上构建左心房模型。
<ol><li>将ICE导管推入股静脉后，穿过下腔静脉和上腔静脉，进入右心房。</li>
	<li>将超声导管推到短轴右心房中间，顺时针旋转。现在，超声波风扇指向描绘右心房和右心室的一点钟方向（家庭视图）。</li>
	<li>拧紧张力 旋钮以实现锁定张力 调整。然后转到 “主页 ”视图并单击 前曲率 （A） 以完全显示三尖瓣环。附着在三尖瓣环上进行门控训练，并使用呼吸末期建模。</li>
	<li>从 主页 视图，顺时针转向左心房前壁，导致左心耳的出现。</li>
	<li>继续顺时针转动至左心房，导致左上肺静脉和下肺静脉的出现，显示为“兔耳征”。然后，顺时针和逆时针转动它，通过识别前金静脉和后金静脉边界来精确收集左肺静脉的图像。</li>
	<li>继续顺时针转动左心房，建立后壁，在此过程中，食管出现“双轨征”。</li>
	<li>顺时针方向转向左心房，观察右下肺静脉，显示为“3字征”。然后，顺时针和逆时针转动它，通过识别前后边界来精确捕获右肺静脉的图像。</li>
	<li>在长轴上，单击 Palintrope（P） 使导管的尖端与冠状窦口的高度相同。这是对左心房模型的补充。调整左弯/ 右弯 （L/R） 以观察左心房长轴的前壁。该图像捕获了左心房的前壁。</li>
	<li>相应地标记重要的解剖位置，包括肺静脉口、左心耳和其他重要部位（视频 1）。</li>
</ol>3.肺静脉区域的图像采集和测量
<ol><li>左心房CT<ol><li>双击图标打开 PACS 系统 。单击 “高级查询 ”以输入患者姓名和检查项目。单击 “确定 ”以查找图像。</li>
		<li>点击 Tune 将图片传输到 vue pacs 系统（图片存档和通信系统）。</li>
		<li>将 3D 体积重建图像传输到工作箱，然后单击导出图像以将左心房后前 （PA）、左后 （LL） 和右外侧 （RL） 位置 图像 保存在文件夹中。</li>
		<li>返回上一个程序后，将左心房动脉期增强序列转移到工作框中，然后单击显示为 3D 的图像。</li>
		<li>双击 3D 图像，然后单击工具栏中的 3D 。选择切除 工具切除 肋骨、脊柱、主动脉和其他结构，以暴露左心房和肺静脉系统。</li>
		<li>暴露肺静脉前庭。单击工具栏中的“ 图 ”，然后选择 “面积 ”以计算肺静脉前庭的横截面积。</li>
	</ol></li>
	<li>冰<ol><li>打开 3D 映射系统。然后，单击 “查看研究 ”并输入患者姓名。最后，使用 “搜索当前患者 ”来识别图像。</li>
		<li>单击 “确定 ”打开工作界面。</li>
		<li>单击 “研究”>“继续研究 ”，然后选择 “模型 ”和 “通道 ”序列。</li>
		<li>单击“ 捕获首选项”，然后选择“ 区域 ”并将图像调整为“后前”、“左外侧”、“右外侧”、“左前斜肌 （LAO）”和“右前斜肌 （RAO）”。</li>
		<li>单击“图片”，选择照片区域，然后单击“确定”以保存图片。</li>
		<li>单击 “ 地图”选项，然后选择 “保存地图”。然后，使用工具栏中的 橡皮擦 去除左右肺静脉。</li>
		<li>单击图像，选择面积测量并测量肺静脉前庭的 面积 。</li>
	</ol></li>
</ol>

基于 ICE 的左心房和肺静脉 3D 重塑

<ol>
	<li>Xu, Y., Sharma, D., Li, G., Liu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atrial+remodeling:+new+pathophysiological+mechanism+of+atrial+fibrillation.">Atrial remodeling: new pathophysiological mechanism of atrial fibrillation.</a> Medical Hypotheses. 80 (1), 53-56 (2013).</li><li>George, J. C., Varghese, V., Mogtader, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracardiac+echocardiography:+evolving+use+in+interventional+cardiology.">Intracardiac echocardiography: evolving use in interventional cardiology.</a> Journal of Ultrasound in Medicine. 33 (3), 387-395 (2014).</li><li>Jingquan, 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=Intracardiac+echocardiography+Chinese+expert+consensus.">Intracardiac echocardiography Chinese expert consensus.</a> Frontiers in Cardiovascular Medicine. 9, 1012731 (2022).</li><li>Anter, E., 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=Comparison+of+intracardiac+echocardiography+and+transesophageal+echocardiography+for+imaging+of+the+right+and+left+atrial+appendages.">Comparison of intracardiac echocardiography and transesophageal echocardiography for imaging of the right and left atrial appendages.</a> Heart Rhythm. 11 (11), 1890-1897 (2014).</li><li>Rossillo, A., 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=Novel+ICE-guided+registration+strategy+for+integration+of+electroanatomical+mapping+with+three-dimensional+CT/MR+images+to+guide+catheter+ablation+of+atrial+fibrillation.">Novel ICE-guided registration strategy for integration of electroanatomical mapping with three-dimensional CT/MR images to guide catheter ablation of atrial fibrillation.</a> Journal of Cardiovascular Electrophysiology. 20 (4), 374-378 (2009).</li><li>Okumura, Y., 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=Three-dimensional+ultrasound+for+image-guided+mapping+and+intervention:+methods,+quantitative+validation,+and+clinical+feasibility+of+a+novel+multimodality+image+mapping+system.">Three-dimensional ultrasound for image-guided mapping and intervention: methods, quantitative validation, and clinical feasibility of a novel multimodality image mapping system.</a> Circulation: Arrhythmia and Electrophysiology. 1 (2), 110-119 (2008).</li><li>Liu, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+evolving+utility+of+intracardiac+echocardiography+in+cardiac+procedures.">The evolving utility of intracardiac echocardiography in cardiac procedures.</a> Journal of Atrial Fibrillation. 6 (6), 1055 (2014).</li><li>Bartel, T., Edris, A., Velik-Salchner, C., M&#252;ller, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracardiac+echocardiography+for+guidance+of+transcatheter+aortic+valve+implantation+under+monitored+sedation:+a+solution+to+a+dilemma.">Intracardiac echocardiography for guidance of transcatheter aortic valve implantation under monitored sedation: a solution to a dilemma.</a> European Heart Journal. Cardiovascular Imaging. 17 (1), 1-8 (2016).</li><li>Haissaguerre, M., 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=Spontaneous+initiation+of+atrial+fibrillation+by+ectopic+beats+originating+in+the+pulmonary+veins.">Spontaneous initiation of atrial fibrillation by ectopic beats originating in the pulmonary veins.</a> New England Journal of Medicine. 339 (10), 659-666 (1998).</li><li>Kaseno, K., 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=The+impact+of+the+CartoSound&#174;+image+directly+acquired+from+the+left+atrium+for+integration+in+atrial+fibrillation+ablation.">The impact of the CartoSound&#174; image directly acquired from the left atrium for integration in atrial fibrillation ablation.</a> Journal of Interventional Cardiac Electrophysiology. 53, 301-308 (2018).</li><li>Enriquez, A., 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=Use+of+intracardiac+echocardiography+in+interventional+cardiology:+working+with+the+anatomy+rather+than+fighting+it.">Use of intracardiac echocardiography in interventional cardiology: working with the anatomy rather than fighting it.</a> Circulation. 137 (21), 2278-2294 (2018).</li></ol>

作者没有利益冲突需要声明。

心内超声心动图（ICE）是一种非接触式三维重建工具。它确定了合适的消融平面，并降低了肺静脉狭窄的发生率。此外，ICE 通过评估消融导管的远端位置及其与解剖结构的相对关联来提高导管消融的有效性。这些结构包括左心房和肺静脉以及肺静脉的直径和形态。
ICE引导的心房颤动导管消融术可以减少术中X线照射，减少房间隔穿刺时间，发现早期并发症，并提供及时治疗，...

心房颤动 （AF） 通常与心房重塑有关，包括机械重塑、电生理重塑和结构重塑1。结构重塑将极大地影响中庭的解剖结构。因此，评估 AF 患者的左心房解剖结构对于 AF 消融手术和任何针对左心房的手术都是必不可少的。对于FAM三维建模，通过不断置换心脏中的磁导管，根据其位置对应于固定磁场的空间位置变化，重建心脏的三维建模。相比之下，ICE 3D 建模通过将传感器定位在 ICE 相控阵导管的头端，将心腔中的二维图像与 3D 电解剖映射系统集成在一起。因此，超声扇区表示 3D 建模，以展示解剖关系和实时导管位置。
根据我们的临床经验，心内超声心动图（ICE）可以识别心房壁边界，并进一步建立3D重塑。然而，在 AF 消融或 3D 重塑期间，大多数 ICE 使用仅提供心房或肺静脉的简要轮廓。最初，ICE用于指导房间隔缺损和卵圆孔未闭2的介入闭合。ICE 可以明确房间隔的位置和形状，并已用于需要房间隔穿刺的各种介入手术3.这些包括心房颤动的射频导管消融术、二尖瓣介入治疗等。 ICE可以精确识别肺静脉边界和心房壁，以建立更详细的3D模型3。目前尚不清楚这种方法是否可以为操作者提供更精确的心房解剖学评估，特别是对于肺静脉、胃窦和经鼻中隔部位。在这项研究中，我们比较了使用传统方法和精确 ICE 程序建立的左心房 CT 图像和 3D 重塑，以确定更多信息。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CARTO V6</td>
			<td>Johnson&#38;Johnson</td>
			<td>6.0.80.45</td>
			<td></td>
		</tr>
		<tr>
			<td>CARTO V7</td>
			<td>Johnson&#38;Johnson</td>
			<td>7.1.80.33</td>
			<td></td>
		</tr>
		<tr>
			<td>PACS system</td>
			<td>Philips(China) Investment Co.,Ltd</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Soundstar</td>
			<td>Johnson&#38;Johnson</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

three-dimensional modeling of the left atrium and pulmonary veins with a precise intracardiac echocardiography approach

心内超声心动图 （ICE） 是一种用于估计肺静脉隔离手术期间心脏解剖结构的新工具，尤其是左心房 （LA） 解剖结构和肺静脉结构。ICE广泛用于在消融过程中建立三维（3D）左心房结构模型。然而，目前尚不清楚在精确的 3D 建模方法中使用 ICE 是否可以提供更准确的左心房 3D 模型和经鼻中隔入路。本研究提出了一种使用 ICE 和快速解剖映射 （FAM） 导管重塑对左心房和肺静脉进行建模的方案。它通过观察者评分来评估使用这两种方法生成的模型的准确性。我们纳入了 50 名接受基于 ICE 的 3D 重塑的患者和 45 名接受基于肺静脉分离程序的 FAM 3D 重塑的患者。肺静脉窦重塑是通过比较重塑和左心房计算机断层扫描血管造影 （CTA） 获得的窦面积来估计的。ICE组和FAM组的观察者得分分别为3.40±0.81分和3.02分±0.72分（P<0.05）。使用基于 ICE 和 FAM 的方法获得的肺静脉窦面积与左心房 CT 获得的面积呈相关性。然而，使用 Bland-Altman 分析，ICE 获取模型的 95% 置信区间偏差比 FAM 获取模型（分别为 -238 cm 2 至 323 cm 2 和 -363 cm 2 至 386 cm 2）更窄。因此，精密ICE在估计左心房结构方面具有很高的准确率，成为未来心脏结构估计的一种有前途的方法。

2021年1月至2022年6月，我们选取了114例在我院接受心房颤动射频消融术的患者。根据以下标准排除患者：无左心房 CT 3D 体积重建图像 （n = 11）、无经隔穿刺 ICE 图像 （n = 4） 以及左心房和肺静脉图像重建不完全 （n = 4）。最后，纳入50例ICE 3D建模患者和45例FAM3D建模患者作为对照组。
两位专业的电生理学家分析了所有的3D建模图像。我们比较了Carto建模和左心房计算机断层扫描?...

Watch this Scientific Journal Video about Advancements in Intracardiac Echocardiography for Atrial Anatomy Assessment at JoVE.com

作者聚焦：用于心房解剖评估的心内超声心动图的进展 

准确的心内超声心动图 （ICE） 在估计左心房结构方面显示出显着的准确性，这是一种有前途且有前途的心脏结构估计方法。在这里，我们提出了一种使用 ICE 和快速解剖映射 （FAM） 导管重塑对左心房和肺静脉进行三维建模的方案。

使用精确的心内超声心动图方法对左心房和肺静脉进行三维建模

Intracardiac echocardiography (ICE) is a novel tool for estimating cardiac anatomy during pulmonary vein isolation procedures, particularly the left ...

我们的研究试图评估基于心内超声心动图的 3D 构建是否可以提高肺心房的准确性和动脉解剖学评估。心内超声心动图能够识别动脉壁边界并创建肺风的 3D 模型。最近，这已经增加，重点是使用S2经常详细的图像来准确确定动脉解剖结构。目前的实验挑战之一是准确模仿左动脉结构。然而，已经获得了有希望的结果，用作最佳的动脉成像和安全解构，具有很高的准确性，并具有未来心脏结构成像的潜力。

three-dimensional-modeling-left-atrium-pulmonary-veins-with-precise

Research

JoVE Journal

Medicine

Three-Dimensional Modeling of the Left Atrium and Pulmonary Veins with a Precise Intracardiac Echocardiography Approach

695 次观看.  China-Japan Union Hospital of Jilin University. 我们的研究试图评估基于心内超声心动图的 3D 构建是否可以提高肺心房的准确性和动脉解剖学评估。心内超声心动图能够识别动脉壁边界并创建肺风的 3D 模型。最近，这已经增加，重点是使用S2经常详细的图像来准确确定动脉解剖结构。目前的实验挑战之一是准确模仿左动脉结构。然而，已经获得了有希望的结果，用作最佳的动脉成像和安全解构，具有很高的准确性，...

视频: 作者聚焦：用于心房解剖评估的心内超声心动图的进展 

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

支气管Thermoplasty：严重哮喘的一种新的治疗方法

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

科研

医学

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical treatments and improved platinum-based chemotherapies, there has been little improvement in EOC survival rates for more than four decades 1,2. Whilst stage I tumors have 5-year survival rates &gt;85%, survival rates for stage III/IV disease are &lt;40%. Thus, the high rates of mortality for EOC could be significantly decreased if tumors were detected at earlier, more treatable, stages 3-5. At present, the molecular genetic and biological basis of early stage disease development is poorly understood. More specifically, little is known about the role of the microenvironment during tumor initiation; but known risk factors for EOCs (e.g. age and parity) suggest that the microenvironment plays a key role in the early genesis of EOCs. We therefore developed three-dimensional heterotypic models of both the normal ovary and of early stage ovarian cancers. For the normal ovary, we co-cultured normal ovarian surface epithelial (IOSE) and normal stromal fibroblast (INOF)&nbsp;cells, immortalized by retrovrial transduction of the catalytic subunit of human telomerase holoenzyme (hTERT) to extend the lifespan of these cells in culture. To model the earliest stages of ovarian epithelial cell transformation, overexpression of the CMYC oncogene in IOSE cells, again co-cultured with INOF cells. These heterotypic models were used to investigate the effects of aging and senescence on the transformation and invasion of epithelial cells. Here we describe the methodological steps in development of these three-dimensional model; these methodologies aren't specific to the development of normal ovary and ovarian cancer tissues, and could be used to study other tissue types where stromal and epithelial cell interactions are a fundamental aspect of the tissue maintenance and disease development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

异型三维<em在体外</em&gt;卵巢癌的发生和发展过程中的间质 - 上皮细胞相互作用模型

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Three-Dimensional (3D) Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in particular (e.g., brain tumors such as glioblastoma multiforme and squamous cell carcinoma of the head and neck &#8211; SCCHN) it is a cause of severe morbidity and can be life-threatening even in the absence of distant metastases. In addition, cancers which have relapsed following treatment unfortunately often present with a more aggressive phenotype. Therefore, there is an opportunity to target the process of invasion to provide novel therapies that could be complementary to standard anti-proliferative agents. Until now, this strategy has been hampered by the lack of robust, reproducible assays suitable for a detailed analysis of invasion and for drug screening. Here we provide a simple micro-plate method (based on uniform, self-assembling 3D tumor spheroids) which has great potential for such studies. We exemplify the assay platform using a human glioblastoma cell line and also an SCCHN model where the development of resistance against targeted epidermal growth factor receptor (EGFR) inhibitors is associated with enhanced matrix-invasive potential. We also provide two alternative methods of semi-automated quantification: one using an imaging cytometer and a second which simply requires standard microscopy and image capture with digital image analysis.

Three-Dimensional 3D Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is a defining characteristic of malignant tumors. We provide here a simple, semi-automated micro-plate assay of invasion into a natural 3D biomatrix that has been exemplified with a number of models of advanced human cancers.

三维（3D）肿瘤球体侵袭试验

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic pressure through echocardiography, as well as to obtain measurements of the Aorta and PA diameter in mice.
Mice older than 10 d of age can be analyzed using the present technique. The technique is composed of 3 main steps: set-up, image acquisition, and image analysis. The set-up step consists of getting the mouse anesthetized with 1% isoflurane, shaving it, and taping it in a supine position to a heated EKG board where the image acquisition will take place. The image acquisition step consists of learning to identify the cardiac structures and obtaining all the required images with its correspondent probe and axis in order to be able to calculate volumes and diameters. The image analysis step consists of measuring the previously acquired images with the aid of computer software.
Advantages of the proposed technique include a fast (15 min) procedure that would allow the researcher to evaluate interventions in a non-invasive, non-terminal approach and therefore follow the same mouse over time; each mouse can be used as its own control. This fact plus having the same operator perform all the acquisition and analysis for the entire experiment minimizes the limitation of operator-dependency. The present methodology is useful for mouse researchers in cardiovascular and pulmonary medicine.

The following protocol describes the methodology for the acquisition and analysis of echocardiographic images used to obtain the Left Atrial Volume (LAV), Aorta (Ao) diameter, and Pulmonary Artery (PA) diameter in mice. This technique is a non-invasive, non-terminal procedure that allows assessment of the cardiopulmonary function.

左心房，主动脉和肺动脉的超声心动图小鼠

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery (ALCAPA)

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ischemia and infarction in children. If left untreated, it results in a 90% mortality rate in the first year of life. In patients who survive to the adulthood, the coronary steal phenomenon and retrograde left-sided coronary flow provide a substrate for chronic subendocardial ischemia, which may lead to left ventricular dysfunction, ischemic mitral regurgitation, malignant ventricular arrhythmias, and sudden cardiac death. The average age of life-threatening presentation is 33 years and of sudden cardiac death 31 years. Therefore, surgical correction is highly recommended as soon as the diagnosis is made, regardless of age. In adult-type ALCAPA originating from the right-facing sinus of the pulmonary artery, direct re-implantation of the ALCAPA into the aorta is the more physiologically sound repair technique to re-establish the dual-coronary perfusion system and is recommended. This protocol describes the technique of direct re-implantation of adult-type ALCAPA into the aorta.

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery ALCAPA

Surgical correction of ALCAPA is highly recommended, regardless of age or the degree of intercoronary collateralization. This protocol presents a technique for the direct re-implantation of adult-type ALCAPA into the aorta to re-establish the dual-coronary perfusion. Whenever feasible, direct re-implantation is preferred to other surgical correction techniques.

左冠状动脉直接再注入与从肺动脉左冠状动脉起源异常成人主动脉（ALCAPA）

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ...

Left Atrial Stenosis Induced Pulmonary Venous Arterialization and Group 2 Pulmonary Hypertension in Rat

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of group 2 PH, due to mitral stenosis (MS), to facilitate the investigation of disease mechanisms and potential therapeutic strategies. We present a novel rat model of pulmonary venous congestion-induced pulmonary venous arterialization and group 2 PH caused by left atrial stenosis (LAS). LAS is achieved by constricting the left atrium using a half-closed titanium clip. After the LAS surgery, a rat model with a transmitral inflow velocity greater than or equal to 2.0 m/s on echocardiography gradually develops pulmonary venous arterialization and group 2 PH over an 8- to 10-week period. In this protocol, we provide the step-by-step procedure of how to perform the LAS surgery. The presented LAS rat model mimics MS in humans and is useful for studying the underlying molecular mechanism of pulmonary venous arterialization and for the preclinical evaluation of therapies for group 2 PH.

Left atrial stenosis (LAS) is a novel surgical technique used for studying group 2 pulmonary hypertension (PH) and mechanisms underlying pulmonary venous arterialization. Here, we present a protocol to constrict the left atrium using a titanium clip to cause pulmonary venous arterialization and moderate PH in a rat.

左心房狭窄致大鼠肺静脉动脉化及第2组肺动脉高压

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of ...

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats with the additional administration of monocrotaline (MCT) or SU5416 (Sugen). We also compare these two models to other PAH models commonly used in research. In the last few years, the focus of animal PAH models has moved towards studying the mechanism of angioproliferation of plexiform lesions, in which the role of increased pulmonary blood flow is considered as an important trigger in the development of severe pulmonary vascular remodeling. One of the most promising rodent models of increased pulmonary flow is the unilateral left pneumonectomy combined with a "second hit" of MCT or Sugen. The removal of the left lung leads to increased and turbulent pulmonary blood flow and vascular remodeling. Currently, there is no detailed procedure of the pneumonectomy surgery in rats. This article details a step-by-step protocol of the pneumonectomy surgical procedure and post-operative care in male Sprague-Dawley rats. Briefly, the animal is anesthetized and the chest is opened. Once the left pulmonary artery, pulmonary vein, and bronchus are visualized, they are ligated and the left lung is removed. The chest then closed and the animal recovered. Blood is forced to circulate only on the right lung. This increased vascular pressure leads to a progressive remodeling and occlusion of small pulmonary arteries. The second hit of MCT or Sugen is used one week post-surgery to induce endothelial dysfunction. The combination of increased blood flow in the lung and endothelial dysfunction produces severe PAH. The primary limitation of this procedure is that it requires general surgical skills.

The rodent left pneumonectomy is a valuable technique in pulmonary hypertension research. Here, we present a protocol to describe the rat pneumonectomy procedure and postoperative care to ensure minimal morbidity and mortality.

左肺切除术联合单氯联氨酸或苏根为大鼠肺动脉高压的模型

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Use of a Percutaneous Ventricular Assist Device/Left Atrium to Femoral Artery Bypass System for Cardiogenic Shock

The left atrial to femoral artery bypass (LAFAB) system is a mechanical circulatory support (MCS) device used in cardiogenic shock (CS) that bypasses the left ventricle by draining blood from the left atrium (LA) and returning it to the systemic arterial circulation via the femoral artery. It can provide flows ranging from 2.5-5 L/min depending on the size of the cannula. Here, we discuss the mechanism of action of LAFAB, available clinical data, indications for its use in cardiogenic shock, steps of implantation, post-procedural care, and complications associated with the use of this device and their management.
We also provide a brief video of the procedural component of device therapy, including the pre-placement preparation, percutaneous placement of the device via transseptal puncture under echocardiographic guidance and the post-operative management of device parameters.

The following article describes the stepwise procedure for placement of a device (e.g., Tandemheart) in cardiogenic shock (CS) that is a percutaneous left ventricular assist device (pLVAD) and a left atrial to femoral artery bypass (LAFAB) system that bypasses and supports the left ventricle (LV) in CS.

使用经皮心室辅助装置/左心房至股动脉旁路系统治疗心源性休克

The left atrial to femoral artery bypass (LAFAB) system is a mechanical circulatory support (MCS) device used in cardiogenic shock (CS) that bypasses the ...

Whole-Kidney Three-Dimensional Staining with CUBIC

Although conventional pathology provided numerous information about kidney microstructure, it was difficult to know the precise structure of blood vessels, proximal tubules, collecting ducts, glomeruli, and sympathetic nerves in the kidney due to the lack of three-dimensional (3D) information. Optical clearing is a good strategy to overcome this big hurdle. Multiple cells in a whole organ can be analyzed at single-cell resolution by combining tissue clearing and 3D imaging technique. However, cell labeling methods for whole-organ imaging remain underdeveloped. In particular, whole-mount organ staining is challenging because of the difficulty in antibody penetration. The present protocol developed a whole-mount mouse kidney staining for 3D imaging with the CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) tissue clearing method. The protocol has enabled visualizing renal sympathetic denervation after ischemia-reperfusion injury and glomerulomegaly in the early stage of diabetic kidney disease from a comprehensive viewpoint. Thus, this technique can lead to new discoveries in kidney research by providing a macroscopic perspective.

The present protocol describes a tissue clearing method and whole-mount immunofluorescent staining for three-dimensional (3D) kidney imaging. This technique can offer macroscopic perspectives in kidney pathology, leading to new biological discoveries.

用立方体进行全肾三维染色

Although conventional pathology provided numerous information about kidney microstructure, it was difficult to know the precise structure of blood ...

使用全肺 3D 重建增强肺结节诊断

Three-Dimensional Reconstruction for the Whole Lung with Early Multiple Pulmonary Nodules

For patients with early multiple pulmonary nodules, it is essential, from a diagnostic perspective, to determine the spatial distribution, size, location, and relationship with surrounding lung tissue of these nodules throughout the entire lung. This is crucial for identifying the primary lesion and developing more scientifically grounded treatment plans for doctors. However, pattern recognition methods based on machine vision are susceptible to false positives and false negatives and, therefore, cannot fully meet clinical demands in this regard. Visualization methods based on maximum intensity projection (MIP) can better illustrate local and individual pulmonary nodules but lack a macroscopic and holistic description of the distribution and spatial features of multiple pulmonary nodules.
Therefore, this study proposes a whole-lung 3D reconstruction method. It extracts the 3D contour of the lung using medical image processing technology against the background of the entire lung and performs 3D reconstruction of the lung, pulmonary artery, and multiple pulmonary nodules in 3D space. This method can comprehensively depict the spatial distribution and radiological features of multiple nodules throughout the entire lung, providing a simple and convenient means of evaluating the diagnosis and prognosis of multiple pulmonary nodules.

作者聚焦：推进 3D 建模以增强早期肺癌肺结节的诊断和治疗 

This study introduces a three-dimensional (3D) reconstruction method for the entire lung in patients with early multiple pulmonary nodules. It offers a comprehensive visualization of nodule distribution and their interplay with lung tissue, simplifying the assessment of diagnosis and prognosis for these patients.

早期多发性肺结节全肺三维重建

For patients with early multiple pulmonary nodules, it is essential, from a diagnostic perspective, to determine the spatial distribution, size, location, ...