我们要感谢内分泌科和手术室小组在这些病人的管理方面给予他们的帮助和支持。我们没有这项工作的资金来源, 也没有利益冲突。

这里的议定书说明了在洛桑大学医院的人类研究伦理委员会的指导方针。
注意: 这里解释的过程集中于对垂体腺瘤进行的经典方法。
1. 术前协议
<ol><li>系统地进行前颅面 CT 扫描和 MRI, 以及内分泌和眼科评估。</li>
</ol>2. 定位和麻醉
<ol><li>诱导全身麻醉并进行口气插管。</li>
	<li>将病人置于全身麻醉和口气插管后的仰卧位。</li>
	<li>将头部定位在头架上, 并固定在头部周围的磁性神经导航带上。</li>
	<li>将表的头部抬高至20°, 以改善静脉引流, 并略微旋转头部约30°对神经外科医生, 同时执行对侧头部倾斜。 注意: 患者头部的伸展或屈曲程度取决于手术靶点: 蝶鞍鞍方法的轻微屈曲 (15-20 °)17 , 对前颅底手术是中性的, 对下斜坡病变可弯曲45°。</li>
	<li>用无酒精的 chlorexidine 溶液消毒脸部。用消毒液清洗鼻腔窝, 然后用棉垫浸渍稀释的因1% 与肾上腺素 (1:1, 000,000)。 注意: 除了局部麻醉和血管收缩外, 这种吸汗还提供了中鼻甲的侧向收缩, 以使内镜的方法。可调节的握臂可固定在手术过程的特定阶段 (特别是在鞍期)。然而, 一些外科医生宁愿不修复内窥镜有一个更动态的视野的外科领域。在这种情况下, 他们要么把内窥镜交给助手, 要么用双手进行工作, 或者把内窥镜本身。</li>
	<li>清洁右腹象限, 并在右侧的围脐区域留下一个小窗口, 同时在术脑脊液瘘的情况下, 它代表一个可能的脂肪接枝收获地点。 注意: 麻醉应特别注意深 morphinic 镇痛, 并在术后的鼻腔阶段控制平均血压, 以减少由于鼻腔粘膜丰富的神经支配而引起的出血。</li>
</ol>3. 手术
注意: 鼻窝一侧的选择主要是由鼻腔解剖 (鼻中隔偏曲, 超鼻甲, 息肉, 既往手术,等) 决定的. 肿瘤的侧向延伸 (侧向伸展), 肿瘤的大小 (binostril 方法用于大肿瘤)。在大多数垂体腺瘤中, 单方面的方法足以使肿瘤及其扩展得到良好的曝光。
<ol><li>
鼻相
	<ol>
<li>介绍0°短刚性内窥镜 (18 厘米长, 4 毫米直径) 进入右鼻孔, 以确定中下鼻甲侧和鼻中隔内侧。喜欢左鼻孔的方法, 以防间隔偏差, 鼻甲异常, 或特定的肿瘤扩展, 在术前成像检测。</li>
		<li>将内窥镜管置于鼻孔的上或下侧, 以便为进入鼻腔的其他仪器提供足够的空间。</li>
		<li>轻轻缩回中间和上级鼻甲与钝刮刀, 以实现广泛的接触同侧蝶窦口。注意支持上级和中间鼻甲通过他们的整个长度与刮刀, 以避免骨折在撤回演习。</li>
	</ol>
</li>
	<li>
蝶相
	<ol>
<li>将鼻粘膜以垂直线性方式凝固, 在蝶骨 osteum 和后之间单约10-15 毫米, 以避免鼻中隔动脉出血 (腭动脉的分支在上缘运行后)。</li>
		<li>垂直打开凝固的鼻粘膜, 直到与犁相对应的骨接触被感觉到。</li>
		<li>跟随犁前直到其与鼻软骨隔膜的交界处, 并把它推向左侧, 露出对侧的蝶骨口, 同时注意保持在粘膜下的平面。</li>
		<li>用骨钳和钳将切除从一个口口延伸到另一口的蝶台上。这个开放的蝶窦允许暴露的 exocranial 颅底面。</li>
		<li>如果蝶骨的黏膜干扰了鞍区的开口, 请轻轻地将其移除。</li>
		<li>切除蝶骨隔膜 (经常有多个隔膜) 与 rongeurs, 以提供广泛的蝶鞍鞍暴露。</li>
		<li>仔细分析术前磁共振成像 (MRI) 和/或 CT 扫描的数量和方向, 这些分隔;这是至关重要的认识到相关的解剖。其中一些分隔可能直接插入颈动脉日珥, 因此应在手术的这一阶段确定为重要的地标 (图 1,图 2,图 3,图 4)。 注: CT 扫描采用64检测器基于系统进行。利用一系列薄切片数据对三维重建进行了处理。MRI 协议是在 3-特斯拉成像仪上进行的。该协议包括一个矢状 T1-weighted 梯度回声序列 (MPRAGE) 与没有对比注射, 160 相邻切片, 1 毫米各向同性体素, 重复时间 (TR) 2300 毫秒, 回声时间 (TE) 2.98 毫秒, 视场 256 mm 为基础的分割,和 3D T2 序列。</li>
	</ol>
</li>
	<li>
鞍期
	<ol>
<li>确定鞍鞍前和优于斜坡休会的中线。 注意: 在大和/或侵袭性垂体肿瘤, 鞍底可能非常薄或完全侵蚀。鞍底是有限的前的抑郁症, 对应于鞍节和横向由颈动脉日珥 (C3 到 C5 段) 和 optico-颈动脉凹陷。</li>
		<li>在侧日珥颈动脉的识别后, 打开鞍底与一个5毫米大的骨刀, 以裁缝或铰链的骨瓣, 可能在手术结束时重新定位。在这一点上, 根据手术目标, 扩大鞍侧截骨或前与骨 rongeurs。 注: 一些外科医生可能更喜欢用菱形钻头打开鞍底地板, 然后用小骨钳扩大骨的张开。此过程可能更快, 并且通常与扩展方法一起使用。然而, 在手术结束时骨不能重新定位, 应注意在中线钻孔以避免颈动脉损伤。使用神经导航来指导过程。</li>
		<li>在侧微的情况下, 在中线为 macroadenoma 或侧面打开鞍硬脑膜。如有需要, 请将此硬脑膜开口侧向, 并注意避免颈动脉损伤。</li>
		<li>通过对硬脑膜进行小的前开口, 并将硬脑膜与吸入尖端一起推上, 以提供足够的通道进入鞍区的内容, 从而限制了前凹槽内蛛网膜破坏的风险。</li>
		<li>用环形角度的刮、镊子和抽吸技巧去除肿瘤组织。</li>
		<li>intra-sellar 生理盐水洗后, 使用30°角内窥镜检查鞍空间, 检查任何残留物, 特别是对海绵窦。</li>
		<li>在切除过程结束前, 要求进行窦机动, 以便在闭合步骤之前检测脑脊液漏。</li>
	</ol>
</li>
	<li>
关闭
	<ol>
<li>如果不是绝对需要, 不要在蝶鞍内放置任何止血材料, 以便更准确地解释术后 MRI。</li>
		<li>在垂体蝶鞍内止血, 用生理盐水冲洗, 用棉垫轻轻按压。</li>
		<li>重建 durotomy 的生物可吸收人工硬脑膜替代物, 被置于 extradurally 和加强它与一薄层的外科粘合剂 (如, bioglue)。</li>
		<li>更换骨瓣, 并用外科粘合剂固定。 注意: 在鞍底被钻孔的情况下, 关节讲台或来自鼻中隔后部的软骨块可用于闭合骨缺损。还介绍了使用合成材料的其他选择 (例如, 如、Porex 板)。</li>
		<li>不要把人工材料留在蝶窦, 以避免任何慢性炎症反应。</li>
		<li>注意鼻腔粘膜的止血, 尤其是两侧腭动脉鼻中隔分支区, 以防止术后出血。</li>
		<li>用钝的刮刀轻轻地将上、中鼻甲替换成原位。</li>
		<li>如果在手术过程中观察到脑脊液漏, 请用小脂肪移植的方法把蝶鞍包起来。除非有必要, 否则不要使用鼻腔包装。</li>
	</ol>
</li>
</ol>4. 术后管理
<ol><li>在围手术期内提供适当的荷尔蒙替代。在前24小时 (100 毫克每天) 使用静脉注射氢化可的松, 然后提供口服替代品 (从病例到病例的变量, 但在住院期间每天至少30毫克)。</li>
	<li>每天两次, 检查血液和尿渗透和电解质, 以及尿密度。持续监测尿量。进行激素检查, 评估术后4天垂体前功能。</li>
	<li>保持阿莫西林/克拉维在1.2 克, 每天两次, 直到5天后。提供标准的止痛治疗, 同时结合醋氨酚和非甾体抗炎药, 提供曲马多止痛。</li>
	<li>经常用生理盐水冲洗鼻窝以限制术后 crostous 鼻炎的风险。</li>
	<li>向病人解释, 避免窦演习的重要性, 以允许正确愈合手术部位在第一个术后几周。</li>
	<li>在术后早期进行 MRI 检查, 如果术中的方面是不典型的垂体腺瘤, 或者怀疑是并发症, 则必须进行手术。术后3月进行脑 MRI 检查。</li>
	<li>在术后3月进行第一次评估, 并与内分泌科合作组织随访, 并根据病理和切除程度进行安排。</li>
</ol>

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Techniques+in+the+Surgical+Treatment+of+Acromegaly:+Applications+and+Efficacy.">Novel Techniques in the Surgical Treatment of Acromegaly: Applications and Efficacy.</a> Neuroendocrinology. 103 (1), 32-41 (2016).</li><li>Shirakawa, M., Yoshimura, S., Yamada, 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=Effectiveness+of+Endoscopic+Endonasal+Transsphenoidal+Surgery+Using+a+Neuronavigation:+Clinical+Results+of+178+Pituitary+Adenomas.">Effectiveness of Endoscopic Endonasal Transsphenoidal Surgery Using a Neuronavigation: Clinical Results of 178 Pituitary Adenomas.</a> Journal of Neurology and Neuroscience. 7 (1:71), (2016).</li><li>Dehdashti, A. R., Ganna, A., Karabatsou, K., Gentili, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pure+endoscopic+endonasal+approach+for+pituitary+adenomas:+early+surgical+results+in+200+patients+and+comparison+with+previous+microsurgical+series.">Pure endoscopic endonasal approach for pituitary adenomas: early surgical results in 200 patients and comparison with previous microsurgical series.</a> Neurosurgery. 62 (5), 1006-1015 (2008).</li><li>Starnoni, D., 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=Surgical+treatment+of+acromegaly+according+to+the+2010+remission+criteria:+systematic+review+and+meta-analysis.">Surgical treatment of acromegaly according to the 2010 remission criteria: systematic review and meta-analysis.</a> Acta Neurochir (Wien). 158 (11), 2109-2121 (2016).</li><li>Alahmadi, H., 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=Impact+of+technique+on+cushing+disease+outcome+using+strict+remission+criteria.">Impact of technique on cushing disease outcome using strict remission criteria.</a> Can J Neurol Sci. 40 (3), 334-341 (2013).</li><li>Donegan, D., 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=Surgical+Outcomes+of+Prolactinomas+in+Recent+Era:+Results+of+a+Heterogenous+Group.">Surgical Outcomes of Prolactinomas in Recent Era: Results of a Heterogenous Group.</a> Endocr Pract. 23 (1), 37-45 (2017).</li><li>Andereggen, L., 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=10-year+follow-up+study+comparing+primary+medical+vs.+surgical+therapy+in+women+with+prolactinomas.">10-year follow-up study comparing primary medical vs. surgical therapy in women with prolactinomas.</a> Endocrine. 55 (1), 223-230 (2017).</li><li>Onnestam, L., 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=National+incidence+and+prevalence+of+TSH-secreting+pituitary+adenomas+in+Sweden.">National incidence and prevalence of TSH-secreting pituitary adenomas in Sweden.</a> J Clin Endocrinol Metab. 98 (2), 626-635 (2013).</li><li>Malchiodi, 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=Thyrotropin-secreting+pituitary+adenomas:+outcome+of+pituitary+surgery+and+irradiation.">Thyrotropin-secreting pituitary adenomas: outcome of pituitary surgery and irradiation.</a> J Clin Endocrinol Metab. 99 (6), 2069-2076 (2014).</li><li>Yamada, 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=Clinicopathological+characteristics+and+therapeutic+outcomes+in+thyrotropin-secreting+pituitary+adenomas:+a+single-center+study+of+90+cases.">Clinicopathological characteristics and therapeutic outcomes in thyrotropin-secreting pituitary adenomas: a single-center study of 90 cases.</a> J Neurosurg. 121 (6), 1462-1473 (2014).</li></ol>

没有利益冲突。

对于大多数垂体肿瘤, 手术仍然是一线治疗的情况下症状患者19,20。仅对泌乳素分泌腺瘤, 第一行治疗是多巴胺激动剂和手术选择次要时, 医疗治疗失败21。
自从在1990代初的22,23中, 内窥镜在颅底手术领域中的引入, 这项技术在大多数医学中心都得到了普遍的应用<sup class="xr...

采用内窥镜技术的颅底手术已明显改善的结果, 颅底手术本身使用或作为辅助的标准显微外科方法。内镜下鼻方法最初介绍治疗鞍区病变1, 但在过去20年中, 这项技术的应用已经扩大, 得益于重大的技术改进和增加的手术经验.内镜下的方法现在也用于前和前侧颅底的病变, 通过延长中线和旁的方法。从前颅底到齿状突的方法现在安全地在许多专门的中心执行, 并取得满意的结果2,3,4,5。
事实上, 内窥镜允许直接可视化病灶通过自然通路, 而不跨越重要的神经血管结构。多项评价对传统 microscope-based 方法的内镜检查程序的安全性和相对优势进行了维持, 并确认主要结局, 如肿瘤切除的程度和激素水平的变化不两种方法不同。然而, 神经并发症, 手术时间和住院长度, 以及病人的术后不适, 明显低于内镜方法6。根据一项挑衅性的出版物, 即使是经验不足的外科医生也可能会在功能垂体腺瘤的队列中使用显微镜技术作为一名非常有经验的外科医生, 获得一个类似的结果.7。
内镜下鼻跨蝶方法实际上是广泛推荐的鞍区病变, 即使在海绵窦侵袭或鞍延长的情况下。如果鞍扩展非常重要, 则可以执行完全内窥的两阶段过程, 同时等待残滓的自然下降8。如果膜片非常紧 ("腰部标志"), 可以执行经颅的方法来解决鞍组件9。关于海绵窦的侵袭, 内窥镜技术似乎优于 Knosp 级2和3功能垂体腺瘤的显微技术10, 而内镜鼻检查可有助于检测侵入海绵窦11。
可能阻碍内镜根治性切除的因素有: 肿瘤延伸到床内动脉, 或 "接吻动脉", 不对称的额延伸, 肿瘤到达 Monro 的孔, 或纤维侵袭周围脑实质的肿瘤9。在这些复杂的病例中, 可以向患者提出经颅结合的方法。此外, 蝶窦的气化是决定手术可行性的一个重要因素, 因为孔沙尔型鼻窦使内窥镜方法更具挑战性12。
通过他们的个人经验约500内镜鼻方法垂体腺瘤, 作者这里描述的不同阶段的方法垂体肿瘤。手术可分为三主要步骤: 鼻, 蝶, 和鞍期。它们是通过完全内镜手术进行的。这里所描述的队列包括微或巨腺瘤的患者, 有一个可变延伸到海绵窦, 蝶窦, 或鞍空间。包括功能和功能腺瘤。
对于其他各种病变, 如咽、垂体转移、斜坡脊索或石油斜坡脑膜瘤, 也可进行全内窥镜手术。然而, 这些病症往往需要更广泛的方法和描述这些变种超出了本文的目的。
指示
这里描述的程序是垂体腺瘤 (功能或功能)。鞍延长不是手术的禁忌, 也不是扩张或侵入海绵窦。情景 CT 扫描分析蝶窦的气化和蝶窦的内隔膜方向。同时, 视神经和颈内动脉的定位是规划手术策略的重要因素。
这种手术技术也可以用于其他肿瘤位于鞍区像咽13, 症状拉特克的裂囊肿, 垂体转移瘤14, 或淋巴瘤15。此外, 其他病变可通过内镜的方法, 如鞍膜脑膜瘤或蝶平原, 石油斜坡区的脑膜瘤和斜坡脊索5,16, 和表皮样或prepontine 池的皮样囊肿。然而, 在这些情况下, 必须采取更广泛的办法。

<table>
	<tbody>
		<tr>
			<td>The StealthStation surgical navigation system</td>
			<td>Medtronic</td>
			<td>ENT Fusion Software 9733652</td>
			<td></td>
		</tr>
		<tr>
			<td>4-mm 18cm 0&#176; rigid endoscope</td>
			<td>Karl Storz</td>
			<td>28132AA</td>
			<td></td>
		</tr>
		<tr>
			<td>4-mm 18cm 30&#176; rigid endoscope</td>
			<td>Karl Storz</td>
			<td>28132BVA</td>
			<td></td>
		</tr>
		<tr>
			<td>4-mm 18cm 45&#176; rigid endoscope</td>
			<td>Karl Storz</td>
			<td>28132FVA</td>
			<td></td>
		</tr>
		<tr>
			<td>Suction Tube</td>
			<td>Karl Storz</td>
			<td>28164XA</td>
			<td></td>
		</tr>
		<tr>
			<td>Coagulation Cannulas</td>
			<td>Karl Storz</td>
			<td>28164MXB</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar Forceps</td>
			<td>Karl Storz</td>
			<td>28164BDB</td>
			<td></td>
		</tr>
		<tr>
			<td>Kerrison Bone Punches</td>
			<td>Karl Storz</td>
			<td>28164MKA</td>
			<td></td>
		</tr>
		<tr>
			<td>Kerrison Bone Punches</td>
			<td>Karl Storz</td>
			<td>28164MKB</td>
			<td></td>
		</tr>
		<tr>
			<td>Kerrison Bone Punches</td>
			<td>Karl Storz</td>
			<td>28164MKC</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164RN</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164RO</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164RI</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164RG</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164GLL</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Curettes</td>
			<td>Karl Storz</td>
			<td>28164GLR</td>
			<td></td>
		</tr>
		<tr>
			<td>de DIVITIS-CAPPABIANCA Scalpels</td>
			<td>Karl Storz</td>
			<td>28164M</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Karl Storz</td>
			<td>28164TD</td>
			<td></td>
		</tr>
		<tr>
			<td>High-speed drill</td>
			<td>Midas Rex Medtronic</td>
			<td>1898430</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic agent</td>
			<td>Floseal, Baxter</td>
			<td>1503350</td>
			<td></td>
		</tr>
		<tr>
			<td>Fibrin sealant</td>
			<td>Tisseel, Baxter</td>
			<td>600065</td>
			<td></td>
		</tr>
		<tr>
			<td>CT Lightspeed</td>
			<td>GE medical systems, Milwaukee, WI, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Carestream Vue PACS</td>
			<td>Carestream Health, Rochester, NY, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>3-Tesla Siemens Tim Trio scanner</td>
			<td>Siemens AG, Erlangen, Germany</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

endoscopic endonasal trans-sphenoidal approach: minimally invasive surgery for pituitary adenomas

内窥镜下鼻跨蝶手术已成为手术治疗垂体腺瘤和许多其他垂体病变的金标准。外科技术的改进, 技术的进步, 以及神经导航的融合使这项手术微创。这一手术的并发症率很低, 而通过这种方法获得的优异结果始终如一。
本文着重介绍了基于个人经验的垂体腺瘤的 step-by 步手术方法, 并详细介绍了这种微创手术所取得的结果。

自2009年以来, 我们的垂体计划已经发展到一个地步, 一个完整的内镜鼻方法已被用于治疗垂体肿瘤的良好术后结果。从2009年到 2016, 两名高级外科医生 (RTD 和 MM) 通过全内窥镜技术对473例垂体腺瘤患者进行手术。组织学在表 1中详细说明: 这些病例中大多数是功能垂体腺瘤 (57%)(图 1), 其次是生长激素 (GH) 分泌腺瘤 (图 4...

Watch this Scientific Journal Video about Endoscopic Endonasal Trans-sphenoidal Approach: Minimally Invasive Surgery for Pituitary Adenomas at JoVE.com

Endoscopic Endonasal Trans-sphenoidal Approach: Minimally Invasive Surgery for Pituitary Adenomas

本文的目的是描述内镜鼻方法对蝶鞍鞍的不同步骤。

内镜下鼻跨蝶入路: 垂体腺瘤的微创手术

Endoscopic endonasal trans-sphenoidal surgery has become the gold standard for the surgical treatment of pituitary adenomas and many other pituitary ...

endoscopic-endonasal-trans-sphenoidal-approach-minimally-invasive

Research

JoVE Journal

Neuroscience

18.7K 次观看.  University Hospital of Lausanne. 本文的目的是描述内镜鼻方法对蝶鞍鞍的不同步骤。

视频: Endoscopic Endonasal Trans-sphenoidal Approach: Minimally Invasive Surgery for Pituitary Adenomas

Minimally-invasive Technique for Injection into Rat Optic Nerve

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.

Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.

微创技术注入到老鼠视神经

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central ...

科研

神经科学

Intramuscular Injections Along the Motor End Plates: A Minimally Invasive Approach to Shuttle Tracers Directly into Motor Neurons

Diseases affecting the integrity of spinal cord motor neurons are amongst the most debilitating neurological conditions. Over the last decades, the development of several animal models of these neuromuscular disorders has provided the scientific community with different therapeutic scenarios aimed at delaying or reversing the progression of these conditions. By taking advantage of the retrograde machinery of neurons, one of these approaches has been to target skeletal muscles in order to shuttle therapeutic genes into corresponding spinal cord motor neurons. Although once promising, the success of such gene delivery approach has been hampered by the sub-optimal number of transduced motor neurons it has so far shown to yield. Motor end plates (MEPs) are highly specialized regions on the skeletal musculature that are in direct synaptic contact to the spinal cord &#945; motor neurons. In this regard, it is important to note that, so far, the efforts to retrogradely transfer genes into motor neurons were made without reference to the location of the MEP region in the targeted muscles. Here, we describe a simple protocol 1) to reveal the exact location of the MEPs on the surface of skeletal muscles and 2) to use this information to guide the intramuscular delivery and subsequent optimal retrograde transport of retrograde tracers into motor neurons. We hope to utilize the results from these tracing experiments in further studies into investigating retrograde transport of therapeutic genes to spinal cord motor neurons through the targeting of MEPs.

The efficacy of intramuscular uptake and retrograde transport of molecules to corresponding motor neurons depends on the location of the injection sites with respect to the motor end plates (MEPs). Here, we describe how to locate MEPs on skeletal muscles to optimise retrograde transport of tracers into motor neurons.

肌肉注射沿运动终板:微创方法来接送示踪剂直接为运动神经元

Diseases affecting the integrity of spinal cord motor neurons are amongst the most debilitating neurological conditions. Over the last decades, the ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific frequency and in turn modulate ongoing cortical oscillatory activity. This neurotool allows the establishment of a causal link between endogenous oscillatory activity and behavior. Most of the tACS studies have shown online effects of tACS. However, little is known about the underlying action mechanisms of this technique because of the AC-induced artifacts on Electroencephalography (EEG) signals. Here we show a unique approach to investigate online physiological frequency-specific effects of tACS of the primary motor cortex (M1) by using single pulse Transcranial Magnetic Stimulation (TMS) to probe cortical excitability changes. In our setup, the TMS coil is placed over the tACS electrode while Motor Evoked Potentials (MEPs) are collected to test the effects of the ongoing M1-tACS. So far, this approach has mainly been used to study the visual and motor systems. However, the current tACS-TMS setup can pave the way for future investigations of cognitive functions. Therefore, we provide a step-by-step manual and video guidelines for the procedure.

Transcranial Alternating Current Stimulation (tACS) allows the modulation of cortical excitability in a frequency-specific fashion. Here we show a unique approach which combines online tACS with single pulse Transcranial Magnetic Stimulation (TMS) in order to "probe" cortical excitability by means of Motor Evoked Potentials.

经颅磁刺激在线联合方法对经颅交流电刺激对原发性运动皮层的影响

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Healthy Brain-pituitary Slices for Electrophysiological Investigations of Pituitary Cells in Teleost Fish

Electrophysiological investigations of pituitary cells have been conducted in numerous vertebrate species, but very few in teleost fish. Among these, the clear majority have been performed on dissociated primary cells. To improve our understanding of how teleost pituitary cells, behave in a more biologically relevant environment, this protocol shows how to prepare viable brain-pituitary slices using the small freshwater fish medaka (Oryzias latipes). Making the brain-pituitary slices, pH and osmolality of all solutions were adjusted to values found in body fluids of freshwater fish living at 25 to 28 &#176;C. Following slice preparation, the protocol demonstrates how to conduct electrophysiological recordings using the perforated whole-cell patch-clamp technique. The patch-clamp technique is a powerful tool with unprecedented temporal resolution and sensitivity, allowing investigation of electrical properties from intact whole cells down to single ion channels. Perforated patch is unique in that it keeps the intracellular environment intact preventing regulatory elements in the cytosol from being diluted by the patch pipette electrode solution. In contrast, when performing traditional whole-cell recordings, it was observed that medaka pituitary cells quickly lose their ability to fire action potentials. Among the various perforation techniques available, this protocol demonstrates how to achieve perforation of the patched membrane using the fungicide Amphotericin B.

The article describes an optimized protocol for making viable brain-pituitary tissue slices, using the teleost fish medaka (Oryzias latipes), followed by electrophysiological recordings of pituitary cells using the patch-clamp technique with the perforated patch configuration.

健康脑-垂体切片对硬骨鱼类鱼垂体细胞电生理的研究

Electrophysiological investigations of pituitary cells have been conducted in numerous vertebrate species, but very few in teleost fish. Among these, the ...

Minimally Invasive Endoscopic Intracerebral Hemorrhage Evacuation

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based treatment options for this devastating disease process. In the past decade, a number of minimally invasive surgeries have emerged to address this issue, one of which is endoscopic evacuation. Stereotactic ICH Underwater Blood Aspiration (SCUBA) is a novel endoscopic evacuation technique performed in a fluid-filled cavity using an aspiration system to provide an additional degree of freedom during the procedure. The SCUBA procedure utilizes a suction device, endoscope, and sheath and is divided into two phases. The first phase involves maximal aspiration and minimal irrigation to decrease clot burden. The second phase involves increasing irrigation for visibility, decreasing aspiration strength for targeted aspiration without disturbing the cavity wall, and cauterizing any bleeding vessels. Using the endoscope and aspiration wand, this technique aims to maximize hematoma evacuation while minimizing collateral damage to the surrounding brain. Advantages of the SCUBA technique include the use of a low-profile endoscopic sheath minimizing brain disruption and improved visualization with a fluid-filled cavity rather than an air-filled one.

This paper details the surgical protocol for minimally invasive endoscopic intracerebral hemorrhage evacuation using the SCUBA technique.

微创内窥镜内腹出血疏散

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based ...

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

Aplysia californica is a model system for studying the neural control of learning and behavior. This animal has a semi-open circulatory system, making it possible to access many of its internal structures without causing any significant damage. Many manipulations can be easily performed both in vivo and in vitro, making it a highly tractable model for the analysis of behavior and neural circuitry. To better understand the functions of muscles within the feeding grasper, we have developed a technique for lesioning them without opening the main body cavity of the animal or damaging the outer layers of the feeding organ (i.e., the buccal mass). In this technique, the grasper is partially everted, allowing direct access to the musculature. This procedure allows animals to recover quickly and reliably. This has made it possible to lesion the I7 muscles and sub-radular fibers, allowing us to show that both muscles significantly contribute to the opening in vivo.

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

Here we present a protocol for minimally invasive surgical lesioning of muscles intrinsic to the feeding apparatus of the marine mollusk Aplysia californica to understand the roles of these muscles during feeding behavior.

阿普利西亚的奥多托福雷内在肌肉的微创病变技术

Aplysia californica is a model system for studying the neural control of learning and behavior. This animal has a semi-open circulatory system, making it ...

Statistical Modelling of Cortical Connectivity Using Non-invasive Electroencephalograms

Non-invasive electrophysiological recordings are useful for the evaluation of nervous system function. These techniques are inexpensive, fast, replicable, and less resource-intensive than imaging. Further, the functional data produced have excellent temporal resolution, which is not achievable with structural imaging.
Current applications of electroencephalograms (EEG) are limited by data processing methods. Standard analysis techniques using raw time series data at individual channels are very limited methods of interrogating nervous system activity. More detailed information about cortical function can be achieved by examining relationships between channels and deriving statistical models of how areas are interacting, allowing visualization of connectivity between networks.
This manuscript describes a method for deriving statistical models of cortical network activity by recording EEG in a standard manner, then examining the interelectrode coherence measures to assess relationships between the recorded areas. Higher order interactions can be further examined by assessing the covariance between the coherence pairs, producing high-dimensional &#34;maps&#34; of network interactions. These data constructs can be examined to assess cortical network function and its relationship to pathology in ways not achievable with traditional techniques.
This approach offers greater sensitivity to network level interactions than is achievable with raw time series analysis. It is, however, limited by the complexity of drawing specific mechanistic conclusions about the underlying neural populations and the high volumes of data generated, requiring more advanced statistical techniques for evaluation, including dimensionality reduction and classifier-based approaches.

Standard EEG analysis techniques offer limited insight into nervous system function. Deriving statistical models of cortical connectivity offers far greater ability to investigate underlying network dynamics. Improved functional assessment opens new possibilities for diagnosis, prognostication, and outcome prediction in nervous system diseases.

使用非侵入性脑电图的皮质连通性统计建模

Non-invasive electrophysiological recordings are useful for the evaluation of nervous system function. These techniques are inexpensive, fast, replicable, ...

A Revised Surgical Approach to Induce Endolymphatic Hydrops in the Guinea Pig

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) remain unclear. In order to adequately study the attributes of endolymphatic hydrops, such as the origins of low-frequency hearing loss, a reliable model is needed. The guinea pig is a&#160;good model because it hears in the low-frequency regions that are putatively affected by endolymphatic hydrops. Previous research has demonstrated that endolymphatic hydrops can be induced surgically via intradural or extradural approaches that involve drilling on the endolymphatic duct and sac. However, whether it was possible to create an endolymphatic hydrops model using an extradural approach that avoided dangerous drilling on the endolymphatic duct and sac was unknown. The objective of this study was to demonstrate a revised extradural approach to induce experimental endolymphatic hydrops at 30 days post-operatively by obliterating the endolymphatic sac and injuring the endolymphatic duct with a fine pick. The sample size consisted of seven guinea pigs. Functional measurements of hearing were made and temporal bones were subsequently harvested for histologic analysis. The approach had a success rate of 86% in achieving endolymphatic hydrops. The risk of cerebrospinal fluid leak was minimal. No perioperative deaths or injuries to the posterior semicircular canal occurred in the sample. The presented method demonstrates a safe and reliable way to induce endolymphatic hydrops at a relatively quick time point of 30 days. The clinical implications are that the presented method provides a reliable model to further explore the origins of low-frequency hearing loss that can be associated endolymphatic hydrops.

This article demonstrates an extradural approach to obliterate the guinea pig endolymphatic sac and injure the endolymphatic duct with a fine pick in order to induce experimental endolymphatic hydrops.

几内亚猪诱导内部水解的修正手术方法

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) ...

Adaptable Angled Stereotactic Approach for Versatile Neuroscience Techniques

Stereotactic surgery is an essential tool in the modern neuroscience lab. However, the ability to precisely and accurately target difficult-to-reach brain regions still presents a challenge, particularly when targeting brain structures along the midline. These challenges include avoiding of the superior sagittal sinus and third ventricle and the ability to consistently target selective and discrete brain nuclei. In addition, more advanced neuroscience techniques (e.g., optogenetics, fiber photometry, and two-photon imaging) rely on targeted implantation of significant hardware to the brain, and spatial limitations are a common hindrance. Presented here is a modifiable protocol for stereotactic targeting of rodent brain structures using an angled coronal approach. It can be adapted to 1) mouse or rat models, 2) various neuroscience techniques, and 3) multiple brain regions. As a representative example, it includes the calculation of stereotactic coordinates for targeting of the mouse hypothalamic ventromedial nucleus (VMN) for an optogenetic inhibition experiment. This procedure begins with the bilateral microinjection of an adeno-associated virus (AAV) encoding a light-sensitive chloride channel (SwiChR++) to a Cre-dependent mouse model, followed by the angled bilateral implantation of fiberoptic cannulae. Using this approach, findings show that activation of a subset of VMN neurons is required for intact glucose counterregulatory responses to insulin-induced hypoglycemia.

Described here is a stereotactic procedure that can target challenging and difficult-to-reach brain regions (due to spatial limitations) using an angled coronal approach. This protocol is adaptable to both mouse and rat models and can be applied to diverse neuroscientific applications, including cannula implantation and microinjections of viral constructs.

多功能神经科学技术的适应性角度立体定向方法

Stereotactic surgery is an essential tool in the modern neuroscience lab. However, the ability to precisely and accurately target difficult-to-reach brain ...

Role of Diffusion MRI Tractography in Endoscopic Endonasal Skull Base Surgery

Endoscopic endonasal surgery has gained a prominent role in the management of complex skull base tumors. It allows the resection of a large group of benign and malignant lesions through a natural anatomical extra-cranial pathway, represented by the nasal cavities, avoiding brain retraction and neurovascular manipulation. This is reflected by the patients' prompt clinical recovery and the low risk of permanent neurological sequelae, representing the main caveat of conventional skull base surgery. This surgery must be tailored to each specific case, considering its features and relationship with surrounding neural structures, mostly based on preoperative neuroimaging. Advanced MRI techniques, such as tractography, have been rarely adopted in skull base surgery due to technical issues: lengthy and complicated processes to generate reliable reconstructions for inclusion in the neuronavigation system. 
This paper aims to present the protocol implemented in the institution and highlights the synergistic collaboration and teamwork between neurosurgeons and the neuroimaging team (neurologists, neuroradiologists, neuropsychologists, physicists, and bioengineers) with the final goal of selecting the optimal treatment for each patient, improving the surgical results and pursuing the advancement of personalized medicine in this field.

We present a protocol to integrate diffusion MRI tractography in patient work-up to endoscopic endonasal surgery for a skull base tumor. The methods for adopting these neuroimaging studies in the pre- and intra-operative phases are described.

扩散MRI造影术在内窥镜内窥镜颅骨基础外科中的作用

Endoscopic endonasal surgery has gained a prominent role in the management of complex skull base tumors. It allows the resection of a large group of ...