这项研究没有得到任何资助。

伊斯坦布尔医学院机构审查委员会批准了该研究方案。在研究开始之前，要求患者签署一份知情同意书，概述其数据的使用目的和发布。
1. 术前程序
<ol><li>在全身麻醉下进行手术。在手术室准备内窥镜和光学设备。</li>
	<li>在开始程序之前检查仪器（图 1 和 表 1）。</li>
</ol>2. 手术技术（图 2）
<ol><li>在全身麻醉下将患者置于仰卧位。用柔软的头部支撑患者头部，并将其弯曲 10° 角。</li>
	<li>触诊冠状缝合线。Kocher 点位于冠状缝合线的正前方，中线右侧约 2.5 cm。</li>
	<li>按照常规无菌和消毒规则开始手术。<ol><li>使用 #5 刀片在右侧额部区域（包括 Kocher 点）做一个 20 厘米的曲线切口。</li>
		<li>通过双极烧灼术确保止血，并解剖皮肤和皮下组织。</li>
		<li>放置自动皮肤牵开器。</li>
		<li>使用 Adson 骨膜电梯刮擦骨膜。</li>
		<li>使用高速钻头和穿孔器在前部区域钻一个毛刺孔。</li>
		<li>用 #11 刀片打开十字形的硬脑膜，并用双极烧灼凝固下面的软脑膜。</li>
		<li>将内窥镜鞘插入垂直于钻孔的脑实质中，并在握住内窥镜鞘的同时取下闭孔。进入侧脑室后，可以看到脑脊液从内窥镜鞘中流出。 注：检查脑脊液 （CSF） 样本中的肿瘤标志物，如 α-胎蛋白和 B-Hcg，可提高组织学检查的准确性。</li>
		<li>将 6.1 mm 宽的内窥镜插入内窥镜护套。可视化脑室侧腔。<ol><li>识别侧脑室中的间隔静脉和丘脑纹静脉。</li>
			<li>识别脉络丛。向前跟随脉络丛直至第三脑室，穿过门罗孔 （FOM）。</li>
		</ol></li>
		<li>将心室镜穿过 FOM 以进入第三脑室。</li>
		<li>找到第三脑室的底部，由于脑积水，它通常很薄。<ol><li>识别第三脑室的体和漏斗部隐窝。</li>
			<li>确保第三脑室底部的孔位于漏斗部隐窝和乳体之间最半透明的点。</li>
		</ol></li>
		<li>找到基底动脉并在动脉复合体前方穿刺，以防止手术过程中的损伤和出血。</li>
		<li>使用 4F Fogarty 导管的尖端刺穿第三脑室的底部。然后，反复给导管的球囊充气和放气，以扩大开口。观察导管球囊放气时脑脊液的射流。</li>
		<li>如果在手术过程中发生出血，请用大量温水冲洗，直到所有出血明显停止并且脑脊液清除。 注意：文献中关于用于管理出血的液体存在不同的看法。但是，我们在诊所使用正常体温的乳酸林格氏液。</li>
		<li>将内窥镜移至松果体肿瘤所在的第三脑室后部区域。 注意：内窥镜的入口点和角度在进入这两个位于不同位置的结构中起着关键作用。这对于避免损坏穹窿和关键血管结构至关重要。</li>
		<li>识别中间马萨并进行解剖，以便更好地观察后第三脑室。</li>
		<li>识别肿瘤并检查生长模式。使用双极烧灼术小心凝固肿瘤表面。</li>
		<li>肿瘤表面完全凝固后，使用活检钳进行活检。将活检送至病理科进行冷冻分析，并根据组织病理学诊断切除肿瘤。</li>
		<li>通过冲洗和双极烧灼来控制局部出血。</li>
		<li>拆下内窥镜系统。</li>
		<li>用缝合线缝合伤口，不要引流管。</li>
	</ol></li>
</ol>3. 术后操作
<ol><li>住院时间最短。手术后第二天动员患者。</li>
	<li>进行头颅 CT 平扫和脑脊液流式 MRI 检查，以评估第三脑室造瘘术的通畅性，并排除出血。</li>
	<li>如果没有主诉，第二天让患者出院。</li>
	<li>要求患者遵循病理结果，并在第 10 天和第一个月末到门诊就诊。</li>
</ol>

Kocher 的内窥镜检查

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M., Yaakub, A., Wan Hitam, W. H., Ahmad Maulana, S. A., Wan Ibrahim, W. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pineal+gland+tumour+with+drop+metastases:+A+case+report.">Pineal gland tumour with drop metastases: A case report.</a> Cureus. 14 (10), e29855 (2022).</li><li>Panyaping, T., Tritanon, O., Wisetsathon, P., Chansakul, T., Pongpitcha, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+apparent+diffusion+coefficient+values+for+distinguishing+between+pineal+germ+cell+tumour+and+pineoblastoma.">Accuracy of apparent diffusion coefficient values for distinguishing between pineal germ cell tumour and pineoblastoma.</a> Clin Radiol. 78 (7), e494-e501 (2023).</li><li>Leone, 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=single+burr+hole+endoscopic+third+ventriculostomy+and+tumor+biopsy+for+pineal+lesions:+Feasibility,+safety,+and+benefits.">single burr hole endoscopic third ventriculostomy and tumor biopsy for pineal lesions: Feasibility, safety, and benefits.</a> World Neurosurg. 173, 5-11 (2023).</li><li>Morgenstern, P. F., 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=Pineal+region+tumors:+An+optimal+approach+for+simultaneous+endoscopic+third+ventriculostomy+and+biopsy.">Pineal region tumors: An optimal approach for simultaneous endoscopic third ventriculostomy and biopsy.</a> Neurosurg Focus. 30 (4), 3 (2011).</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=Management+of+hydrocephalus+secondary+to+pineal+region+tumors.">Management of hydrocephalus secondary to pineal region tumors.</a> Clin Neurol Neurosurg. 115 (9), 1809-1813 (2013).</li><li>Xin, C., 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=Endoscopic-assisted+surgery+versus+microsurgery+for+pineal+region+tumors:+A+single-center+retrospective+study.">Endoscopic-assisted surgery versus microsurgery for pineal region tumors: A single-center retrospective study.</a> Neurosurg Rev. 44 (2), 1017-1022 (2021).</li><li>Choque-Velasquez, J., Resendiz-Nieves, J., Colasanti, R., Hernesniemi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+obstructive+hydrocephalus+associated+with+pineal+region+cysts+and+tumors+and+its+implication+in+long-term+outcome.">Management of obstructive hydrocephalus associated with pineal region cysts and tumors and its implication in long-term outcome.</a> World Neurosurg. 149, e913-e923 (2021).</li><li>Chibbaro, 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=Neuroendoscopic+management+of+posterior+third+ventricle+and+pineal+region+tumors:+Technique,+limitation,+and+possible+complication+avoidance.">Neuroendoscopic management of posterior third ventricle and pineal region tumors: Technique, limitation, and possible complication avoidance.</a> Neurosurg Rev. 35 (3), 331-338 (2012).</li><li>Ahmed, A. 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作者声明他们没有利益冲突。

松果体瘤的 ETV 和活检的描述最早是在 1970 年代描述的。从历史上看，对失控出血的恐惧一直存在。然而，由于内窥镜手术技术的进步，出血控制现在并不是经验丰富的外科医生的主要并发症 9,10。根据文献中的几个案例，已发现内窥镜管理作为表现为脑积水的松果体区域肿瘤的第一步是有效的。内窥镜管理的主要目标是 CSF 引流和组织诊断，以便进一步管理8。
松果体瘤是一组具有不同组织学特征的多种肿瘤。肿瘤可分为四大类：生殖细胞肿瘤、松果体来源的肿瘤、神经上皮来源的肿瘤和各种其他肿瘤，包括转移瘤。考虑到这些肿瘤可用的治疗方案范围广泛以及与完全切除相关的潜在风险，在诊断时使用微创组织取样准确诊断肿瘤非常重要。继发于松果体肿瘤的急性脑积水患者需要立即治疗脑积水，微创内窥镜手术提供了同时诊断组织学的机会 1,7。
ETV 同时对松果体病变进行活检已被用作这些罕见肿瘤的管理策略 6,11,12,13。这种方法有几个优点，包括除 CSF 引流外的活检取样、用于研究肿瘤标志物（如 α-胎蛋白和人绒毛膜促性腺激素）的 CSF 取样，以及能够在直接观察下最大限度地减少丰富血管组织的出血 7,14,15。该区域的肿瘤类型表现出广泛的多样性，并且每种肿瘤内可能存在异质性，例如畸胎瘤、松果细胞瘤和神经胶质瘤。因此，要确定正式的治疗策略，准确的诊断是必不可少的 1,6,16。
内窥镜松果体区活检期间最常见的两种术中并发症是静脉出血引起的脑室内出血和狭窄孔 Monro17 患者使用硬质内窥镜引起的穹窿医源性挫伤。随着内窥镜手术、术前计划和适当患者选择经验的增加，这些并发症的发生率显著降低。当单个入路部位用于 ETV 和松果体活检时，使用带有 30° 角镜片的脑室镜可提供更宽的视野和更少的穹窿外翻，从而减少并发症18。
对于在单端口手术中肿瘤活检前进行 ETV 的最佳方法尚未达成共识。选择脑室造瘘术作为第一步的基本原理是，由于活检后有肿瘤出血的风险，视觉模糊的可能性可能会使 ETV 复杂化。对于颅内压极高和梗阻性脑积水的患者，应优先治疗脑积水17。
ETV 和脑室腹腔分流放置等各种技术已用于治疗与松果体区肿瘤相关的阻塞性脑积水19。许多患者在化疗和放疗切除病灶或缩小病灶后可能不需要 CSF 分流术 20。对于与松果体区病变相关的梗阻性脑积水，ETV 是一种广泛使用的微创技术。此外，使用 ETV 获得的肿瘤活检，尤其是用于诊断生殖细胞肿瘤的肿瘤活检，由于其高放射敏感性，可以避免手术切除21。
内窥镜活检和第三脑室造瘘术可以使用单门技术在一次会话中使用刚性和柔性内窥镜进行11。然而，柔性内窥镜系统的光学质量较差已成为一个重大限制，可能会阻碍检测肿瘤扩散的能力22。此外，与刚性内窥镜镊子相比，柔性镊子的尺寸较小，可能会影响活检样本的大小，从而导致组织学结果不一致10。此外，可能难以操纵柔性脑室镜穿过门罗孔并朝向第三脑室的马萨中间体。此外，应在不冲洗的情况下对肿瘤进行活检和凝固。更重要的是，活检部位大量出血可能导致视力完全丧失10,11。
总之，这些肿瘤的治疗方案各不相同。由于完全切除靠近重要的解剖结构，因此具有较高的发病率和死亡率风险。如果患者出现由松果体肿瘤引起的急性阻塞性脑积水，则必须及时治疗脑积水。内窥镜方法提供了同时组织学诊断的可能性23。

松果体是位于上丘脑的神经内分泌腺，负责调节脊椎动物的生物节律。其解剖边界包括形成腺体底部的第三脑室壁的后表面、上部的胼胝体脾和围绕两侧的丘脑。它呈松果状，在四叉心池 1,2 中向后和向下延伸。
松果体肿瘤是相对罕见的肿瘤，主要是儿童恶性肿瘤，占所有儿童脑肿瘤的 3-11%，而成人脑肿瘤的 <1% 1,3,4。年龄、性别和种族可以改变松果体肿瘤的相对发病率5。松果体瘤分为三种类型：生殖细胞瘤、松果体实质肿瘤和起源于邻近解剖区域的肿瘤。生殖细胞瘤是最常见的松果体瘤，占欧洲松果体瘤的 50% 1,3。
松果体肿瘤的病理学多种多样，其最佳管理仍存在争议6。神经内窥镜检查的进步为松果体区肿瘤的治疗做出了重大贡献。神经内窥镜技术在治疗这些肿瘤方面是微创、有效且安全的。使用这种技术，可以治疗脑积水并同时进行活检7。在大多数情况下，由于放疗和化疗对大多数组织病理学亚型有效，同时内镜下第三脑室造瘘术 （ETV） 和肿瘤活检是松果体区肿瘤的首选手术方法。已经描述了同时进行 ETV 和活检的各种技术，目前没有执行该程序的标准技术8。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adson periosteal elevator</td>
			<td>Ruggles-Redmond</td>
			<td>RO263</td>
			<td>Semi-sharp, 5 mm, curved 6-3/8, length 164 mm</td>
		</tr>
		<tr>
			<td>Automatic skin retractors&#160;</td>
			<td>Integra</td>
			<td>3,72,245</td>
			<td>Heiss Automatic Skin Retractor Length - Overall (mm): 102 
			Length 1 - Tip/Jaw (mm): 8</td>
		</tr>
		<tr>
			<td>Balloon catheter&#160;</td>
			<td>Edwards Fogarty</td>
			<td>120804FP</td>
			<td>Length (cm): 80, Catheter size (F): 4,&#160; Inflated balloon diameter (mm): 9</td>
		</tr>
		<tr>
			<td>Biopsy Forceps</td>
			<td>Karl Storz LOTTA CLICKLINE Grasping Forceps</td>
			<td>28164 LE</td>
			<td>Rotating, dismantling, single action jaws, diameter 2.7 mm, working length 30 cm</td>
		</tr>
		<tr>
			<td>Bipolar coagulation electrode</td>
			<td>Karl Storz LOTTA</td>
			<td>28161 SF</td>
			<td>Diameter 1.3 mm, working length 30 cm</td>
		</tr>
		<tr>
			<td>Bisture</td>
			<td>Beybi</td>
			<td>24,02,502</td>
			<td>Beybi Bisture Tip. No: 20 and No: 11</td>
		</tr>
		<tr>
			<td>High-speed drill</td>
			<td>Medtronic Midas Rex MR8</td>
			<td>MR8&#8482; Electric Plus EM850</td>
			<td>Perforator tip used</td>
		</tr>
		<tr>
			<td>Obturator</td>
			<td>Karl Storz LOTTA</td>
			<td>28164 LLO</td>
			<td>Use with Operating Sheaths for ventricular puncture</td>
		</tr>
		<tr>
			<td>Operating sheath</td>
			<td>Karl Storz LOTTA</td>
			<td>28164 LSB</td>
			<td>Graduated, rotating, outer diameter 6.8 mm, working length 13 cm</td>
		</tr>
		<tr>
			<td>Ventriculoscope</td>
			<td>Karl Storz LOTTA Ventriculoscope with HOPKINS</td>
			<td>28164 LAB</td>
			<td>Wide angle telescope 30&#176;, angled eyepiece, outer diameter 6.1 mm, length 18 cm, working channel diameter 2.9 mm, irrigation/suction channel diameter 1.6 mm</td>
		</tr>
	</tbody>
</table>

endoscopic third ventriculostomy and pineal biopsy from a single entry point

松果体肿瘤对儿童有重大影响，尽管它们相对少见。它们约占所有儿童脑肿瘤的 3-11%，远高于成人脑肿瘤的 <1%。这些肿瘤可分为三大类：生殖细胞肿瘤、实质松果体肿瘤和由相关解剖结构引起的肿瘤。获得准确的微创组织诊断对于为松果体肿瘤患者选择最合适的治疗方案至关重要。这是由于可用的治疗方案多种多样，以及与完全切除相关的潜在风险。如果患者出现由松果体肿瘤引起的急性梗阻性脑积水，则必须立即治疗脑积水。这种紧迫性源于脑积水的潜在并发症，包括颅内压升高和神经功能缺损。为了应对这些挑战，微创内窥镜方法提供了一个宝贵的机会。该技术使临床医生能够及时缓解脑积水并同时获得组织学诊断。这种双重好处使患者能够更全面地了解肿瘤，并有助于确定对患者最有效的治疗策略。

术前磁共振成像 （MRI） 显示松果体瘤和三心室脑积水。手术前，我们确保检查了我们的内窥镜装置、4F Fogarty 球囊导管 （见 图 1）和所有必需的材料 （见表 1）。 松果体肿瘤的大小最初测量为 30 mm x 15 mm x 20 mm。然而，在术后 MRI 扫描中，观察到肿瘤大小增加到 35 mm x 52 mm x 45 mm，表明肿瘤具有进行性。幸运的是，在接受化疗后，对治疗有积极的反应。肿瘤大小显著缩小至 15 mm x 15 mm x 10 mm，表明结果成功。
此外，造影剂摄取也显著降低，进一步支持化疗的有效性。 图 3 提供了整个治疗期间肿瘤大小和脑积水减少的直观表示。该图形表示突出了在管理肿瘤和脑积水方面取得的积极进展。在外科手术过程中，采用连续冲洗，因此难以准确测量失血量。但值得注意的是，该患者术中不需要输血，表明失血不显著。
此外，手术后血红蛋白水平没有显着降低，表明患者的血液水平在整个手术过程中保持稳定。总体而言，化疗和手术干预的结合已被证明可有效减小松果体肿瘤的大小和控制三心室脑积水。对治疗的积极反应在肿瘤大小和造影剂摄取的减少中显而易见，为患者的康复提供了希望。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/66837/66837fig01.jpg"> 图 1：Karl-Storz 内窥镜套件和 4F Fogarty 球囊导管。1： 活检钳;2： 内窥镜护套; 3： 闭孔器; 4： 心室镜; 5： 双极凝血电极; 6： 4F 球囊导管。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66837/66837fig01large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/66837/66837fig02.jpg"> 图 2：使用 Kocher 点使用硬质内窥镜进行内窥镜第三脑室造瘘术和松果体活检的示意图。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66837/66837fig02large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/66837/66837fig03.jpg"> 图 3：对松果体肿瘤和三心室脑积水患者进行的磁共振成像扫描。 （A） 术前对比增强 T1 序列矢状面;（B） 术前对比增强 T1 序列轴向;（C） 术前 T2 序列矢状面;（D） 术前 T2 序列轴向。（E） 化疗前和手术后 1 个月对比增强 T1 序列矢状面 MRI;（F） 化疗前和手术后 1 个月对比增强 T1 序列轴向 MRI。（G） 化疗前和手术后 1 个月的 T2 序列矢状面 MRI;（H） 化疗前和手术后 1 个月的 T2 序列轴向 MRI。（I） 化疗后第 2 个月对比增强 T1 序列矢状面 MRI;（J） 化疗后第 2 个月和轴向增强 T1 序列轴向 MRI。（K） 化疗后第 2 个月和轴向 T2 序列矢状面 MRI;（L） 化疗后第 2 个月的 T2 序列轴向 MRI。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66837/66837fig03large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>

作者聚焦：用于第三脑室造口术和松果体活检的单入口内窥镜脑室内入路

松果体肿瘤经常引起梗阻性脑积水，需要组织病理学诊断来决定治疗方案。他们可能会根据病理诊断通过手术切除或放化疗进行治疗。作为一种微创初始治疗，Kocher 点的内窥镜方法可实现第三脑室造瘘术和活检。

内窥镜下第三脑室造口术和松果体活检从单一入口点

Pineal neoplasms have a significant impact on children although they are relatively uncommon. They account for approximately 3-11% of all childhood brain ...

endoscopic-third-ventriculostomy-pineal-biopsy-from-single-entry

Nanyang Technological University

Research

JoVE Journal

Medicine

Endoscopic Third Ventriculostomy and Pineal Biopsy from a Single Entry Point

472 次观看.  Istanbul University Faculty of Medicine. 内窥镜脑室内入路广泛用于神经外科。在这里，我们描述了从单个入口点对因松果体病变患有梗阻性脑积水的患者进行第三脑室造口术和松果体活检的完整内窥镜技术。所以内窥镜系统包含一个通道和一个冲洗通道。双极凝血、神经球囊和微镊子足以进行手术。先进技术的优势使在内窥镜手术中能够采集高清 HDI 4K 质量的图像。倾斜和柔性内窥?...

视频: 作者聚焦：用于第三脑室造口术和松果体活检的单入口内窥镜脑室内入路

Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling

The generation of patient-specific cell lines represents an invaluable tool for diagnostic or translational research, and these cells can be collected from skin or muscle biopsy tissue available during the patient&#8217;s diagnostic workup. In this protocol, we describe a technique for live cell isolation from small amounts of muscle or skin tissue for primary cell culture. Additionally, we provide a technique for the immortalization of myogenic cell lines and fibroblast cell lines from primary cells. Once cell lines are immortalized, substantial expansion of patient-derived cells can be achieved. Immortalized cells are amenable to many downstream applications, including drug screening and in vitro correction of the genetic mutation. Altogether, these protocols provide a reliable tool to generate and preserve patient-derived cells for downstream applications.

This protocol describes techniques for live cell isolation and primary culture of myogenic and fibroblast cell lines from muscle or skin tissue. A technique for the immortalization of these cell lines is also described. Altogether, these protocols provide a reliable tool to generate and preserve patient-derived cells for downstream applications.

从肌肉活检患者来源的细胞系疾病建模隔离和永生化

The generation of patient-specific cell lines represents an invaluable tool for diagnostic or translational research, and these cells can be collected ...

科研

医学

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

线粒体的制备及透气性评估，从骨骼肌组织通过穿刺活检获得的分离

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy

The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem cells (ISCs) and progeny that progressively migrate and differentiate toward the tip of the villi. These processes, essential for gastrointestinal homeostasis, have been extensively studied using multiple approaches. Ex vivo technologies, especially primary cell cultures have proven to be promising for understanding intestinal epithelial functions. A long-term primary culture system for mouse intestinal crypts has been established to generate 3-dimensional epithelial organoids. These epithelial structures contain crypt- and villus-like domains reminiscent of normal gut epithelium. Commonly, termed &#8220;enteroids&#8221; when derived from small intestine and &#8220;colonoids&#8221; when derived from colon, they are different from organoids that also contain mesenchyme tissue. Additionally, these enteroids/colonoids continuously produce all cell types found normally within the intestinal epithelium. This in vitro organ-like culture system is rapidly becoming the new gold standard for investigation of intestinal stem cell biology and epithelial cell physiology. This technology has been recently transferred to the study of human gut. The establishment of human derived epithelial enteroids and colonoids from small intestine and colon has been possible through the utilization of specific culture media that allow their growth and maintenance over time. Here, we describe a method to establish a small intestinal and colon crypt-derived system from human whole tissue or biopsies. We emphasize the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids.

We describe a method to establish human enteroids from small intestinal crypts and colonoids from colon crypts collected from both surgical tissue and biopsies. In this methodological article, we present the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids.

从整体组织和活检建立人类上皮Enteroids和Colonoids的

The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem ...

A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To obtain subcutaneous adipose tissue biopsies in the abdominal area, researchers and physicians use either a surgical or a needle-based technique. However, surgical subcutaneous fat biopsies can offer tissue samples that may provide a more comprehensive overview of the complexities of biological indices in white adipose tissue. Usually, a surgical adipose tissue biopsy includes a diathermy treatment for cauterizing blood vessels to prevent excessive bleeding. Nevertheless, side effects, such as flash fires and skin lesions in the tissue, have been reported after diathermy. Therefore, we aimed to standardize a surgical abdominal adipose tissue biopsy performed under local anesthesia using a non-diathermy method. We conducted 115 subcutaneous adipose tissue biopsies in healthy men using a non-diathermy abdominal surgical biopsy method. Our results showed three cases of excessive post-operation bleeding out of 115 operations (2.61%).In conclusion, our standardized subcutaneous abdominal adipose tissue surgical biopsy using a non-diathermy method can be safely applied to healthy men at the bedside, with minimal side effects.

We standardized an abdominal adipose tissue biopsy using a non-diathermy method performed under local anesthesia. Three cases of excessive post-operation bleeding out of 115 operations (2.61%) occurred.We conclude that an abdominal adipose tissue surgical biopsy using a non-diathermy method can be safely applied to healthy men.

经非透法皮下腹脂肪组织活检技术

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

使用化学活检进行移植质量评估

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...

Endaural Endoscopic Atticoantrotomy (Retrograde Mastoidectomy) using a Constant Suction Bone-drilling Technique

Endoscopic middle ear surgery is a widely employed minimally invasive surgical technique to address middle ear and mastoid pathology. Bone drilling is the main technical challenge of endoscopic middle ear surgery. The accompanying video describes the detailed protocol of a constant-suction bone-drilling technique and the procedure of endaural exclusive endoscopic atticoantrotomy (retrograde mastoidectomy) using this technique. The main components of this bone-drilling technique include a soft and flexible suction tube, which is placed into the tympanic cavity to provide constant suction, and a soft sleeve, which is wrapped around the drill shaft to prevent the high-speed rotating shaft from damaging the lens of the endoscope. With these simple modifications, the traditional otological electrodrill can be used for drilling a tiny endaural incision in endoscopic middle ear surgery. Based on this bone-drilling technique, endaural endoscopic atticoantrotomy (retrograde mastoidectomy) can be successfully established for the removal of various amounts of bone, depending on the extent of the lesion. The short-term postoperative outcome seems promising.

Endaural Endoscopic Atticoantrotomy Retrograde Mastoidectomy using a Constant Suction Bone-drilling Technique

In this paper, we present the protocol and short-term outcomes of endaural exclusive endoscopic atticoantrotomy using a constant suction bone-drilling technique.

使用恒定抽吸骨术的内窥镜下盂骨切开术（逆行切除术）

Endoscopic middle ear surgery is a widely employed minimally invasive surgical technique to address middle ear and mastoid pathology. Bone drilling is the ...

Endoscopic Ultrasound-Guided Biliary Drainage: Endoscopic Ultrasound-Guided Hepaticogastrostomy in Malignant Biliary Obstruction

Patients with unresectable malignant biliary obstruction often require biliary drainage to decompress the biliary system. Endoscopic Retrograde Cholangiopancreatography (ERCP) is the primary biliary drainage method whenever possible. Percutaneous Transhepatic Biliary Drainage (PTBD) is used as a salvage method if ERCP fails. Endoscopic Ultrasound-Guided Biliary Drainage (EUS-BD) provides a feasible alternative biliary drainage method where one of the methods is EUS guided Hepaticogastrostomy (EUS-HGS). Here we describe the EUS-HGS technique in a case of unresectable malignant hilar biliary obstruction to achieve biliary drainage.
Presented here is the case of a 71-year-old female with painless jaundice and weight loss for 2 weeks. Computed Tomography (CT) imaging showed a 4 x 5 cm hilar tumor with lymphadenopathy and liver metastasis. EUS fine needle biopsy (FNB) of the lesion was consistent with cholangiocarcinoma. Her bilirubin levels were 212 &#181;mol/L (&lt;15) during presentation.
A linear echoendoscope was used to locate the left dilated intrahepatic ducts (IHD) of the liver. The segment 3 dilated IHD was identified and punctured using a 19 G needle. Contrast was used to opacify the IHDs under fluoroscopic guidance. The IHD was cannulated using a 0.025-inch guidewire. This was followed by the dilation of the fistula tract using a 6 Fr electrocautery dilator along with a 4 mm biliary balloon dilator. A partially covered metallic stent of 10 cm in length was deployed under fluoroscopic guidance. The distal part opens in the IHD and the proximal part was deployed within the working channel of the echoendoscope that subsequently released into the stomach. The patient was discharged three days after the procedure. Follow up performed in the second and fourth weeks showed that the bilirubin levels were 30 &#181;mol/L and 14 &#181;mol/L, respectively. This indicates that EUS-HGS is a safe method for biliary drainage in unresectable malignant biliary obstruction.

Endoscopic Ultrasound-Guided Biliary Drainage (EUS-BD) is an alternative method of biliary decompression in malignant biliary obstruction. Here we describe the technique of EUS guided-Hepaticogastrostomy (EUS-HGS) in a case of unresectable malignant hilar biliary obstruction.

内镜超声引导下胆道引流:内镜超声引导下肝胃造瘘术治疗恶性胆道梗阻

Patients with unresectable malignant biliary obstruction often require biliary drainage to decompress the biliary system. Endoscopic Retrograde ...

Endoscopic Cholesteatoma Surgery

Implementation of endoscopes in cholesteatoma surgery resulted in considerable changes in the management of cholesteatoma in the last two decades. Compared to the microscopic approach with an excellent but straight-line view and limited illumination, the introduction of endoscopes provides a wide-angled panoramic view. Moreover, angled lenses allow the surgeon to visualize the middle ear and its hidden recesses through a transcanal, minimally-invasive approach. The endoscope enables the surgeon to remove limited cholesteatoma of the middle ear and its recesses using an exclusive endoscopic technique by taking advantage of these benefits. This reduces the rate of residual disease and sparing external incisions and excessive temporal bone drilling as in a transmastoid approach. Since transcanal endoscopic access is mainly a one-handed technique, it implies the need for specific procedures and technical refinements. This article describes a step-by-step guide as a surgical manual for endoscopic removal of epitympanic cholesteatoma. Different techniques for cholesteatoma dissection and bone removal for epitympanectomy, including curettage and powered instruments such as drills and ultrasonic devices with their outcomes, are discussed. This may offer ear surgeons insight into technical refinements and the latest technological developments and open the horizon for different techniques.

The present protocol describes a step-by-step guide for the complete endoscopic removal of epitympanic cholesteatoma with different techniques for cholesteatoma dissection and bone removal for epitympanectomy.

内窥镜胆脂瘤手术

Implementation of endoscopes in cholesteatoma surgery resulted in considerable changes in the management of cholesteatoma in the last two decades. ...

超声评估膈肌功能障碍和监测治疗

Measuring Diaphragm Thickness and Function Using Point-of-Care Ultrasound

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes affected individuals to respiratory failure. In mechanically ventilated patients, the diaphragm is susceptible to atrophy and dysfunction through disuse and other mechanisms. This contributes to failure to wean and poor long-term clinical outcomes. Point-of-care ultrasound provides a valid and reproducible method for evaluating diaphragm thickness and contractile activity (thickening fraction during inspiration) that can be readily employed by clinicians and researchers alike. This article presents best practices for measuring diaphragm thickness and quantifying diaphragm thickening during tidal breathing or maximal inspiration. Once mastered, this technique can be used to diagnose and prognosticate diaphragm dysfunction, and guide and monitor response to treatment over time in both healthy individuals and acute or chronically ill patients.

作者聚焦:揭示机械通气对隔膜功能和患者预后的影响 

Diaphragm thickness and function can be assessed in healthy individuals and critically ill patients using point-of-care ultrasound. This technique offers an accurate, reproducible, feasible, and well-tolerated method for evaluating diaphragm structure and function.

使用床旁超声测量隔膜厚度和功能

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes ...

内窥镜下第三脑室造口术和松果体活检从单一入口点

副本

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探索更多视频

此视频中的章节

从肌肉活检患者来源的细胞系疾病建模隔离和永生化

线粒体的制备及透气性评估，从骨骼肌组织通过穿刺活检获得的分离

从整体组织和活检建立人类上皮Enteroids和Colonoids的

经非透法皮下腹脂肪组织活检技术

使用化学活检进行移植质量评估

使用恒定抽吸骨术的内窥镜下盂骨切开术（逆行切除术）

内镜超声引导下胆道引流:内镜超声引导下肝胃造瘘术治疗恶性胆道梗阻

内窥镜胆脂瘤手术

使用床旁超声测量隔膜厚度和功能

使用床旁超声测量隔膜厚度和功能