Name: a revised surgical approach to induce endolymphatic hydrops in the guinea pig
Uploaded: 2020-06-04T13:30:00
Description: Watch this Scientific Journal Video about A Revised Surgical Approach to Induce Endolymphatic Hydrops in the Guinea Pig at JoVE.com

我们感谢香农·勒夫勒在数字和材料表方面给予的帮助。本出版物中报告的研究得到了国家卫生研究院内国家耳聋和其他通信障碍研究所的支持，通过"学术耳鼻喉科临床医生/研究人员的发展"培训补助金，奖励编号T32DC000022（C.V.V.）和R01 DC014997（J.T.L.）。内容完全由作者负责，不一定代表国家卫生研究院的官方观点。

下文在《议定书》部分所列的所有程序均按圣路易斯华盛顿大学机构动物护理和使用委员会批准的议定书所述进行。
1. 生命体征的麻醉诱导和监测
注：这项研究使用了从内部繁殖群获得的色素NIH菌株豚鼠。
<ol><li>使用任何性别的豚鼠，体重至少350克。</li>
	<li>将豚鼠放入新生儿暖化异体中，并给予氯胺酮/锡兰辛混合物内腹（50毫克/千克氯?...

<ol>
	<li>Kimura, R. S., Schuknecht, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membranous+Hydrops+in+the+Inner+Ear+of+the+Guinea+Pig+after+Obliteration+of+the+Endolymphatic+Sac.">Membranous Hydrops in the Inner Ear of the Guinea Pig after Obliteration of the Endolymphatic Sac.</a> Pract oto-rhino-laryng. 27, 343-354 (1965).</li><li>Salt, A. N., DeMott, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+of+endolymph+volume+increase+in+experimental+hydrops+measured+in+vivo+with+an+ionic+volume+marker.">Time course of endolymph volume increase in experimental hydrops measured in vivo with an ionic volume marker.</a> Hearing Research. 74 (1-2), 165-172 (1994).</li><li>Walsted, A., Garbarsch, C., Michaels, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+craniotomy+and+cerebrospinal+fluid+loss+on+the+inner+ear.+An+experimental+study.">Effect of craniotomy and cerebrospinal fluid loss on the inner ear. An experimental study.</a> Acta Oto-Laryngologica. 114 (6), 626-631 (1994).</li><li>Andrews, J. C., Bohmer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+surgical+approach+to+the+endolymphatic+sac+and+the+cochlear+aqueduct+in+the+guinea+pig.">The surgical approach to the endolymphatic sac and the cochlear aqueduct in the guinea pig.</a> American Journal of Otolaryngology. 10 (1), 61-66 (1989).</li><li>Lee, J. R., Wright, C. G., Meyerhoff, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modified+occipital+approach+to+the+endolymphatic+sac+and+cochlear+aqueduct+of+the+guinea+pig.">Modified occipital approach to the endolymphatic sac and cochlear aqueduct of the guinea pig.</a> American Journal of Otolaryngology. 14 (2), 165-169 (1993).</li><li>Megerian, C. 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=Surgical+induction+of+endolymphatic+hydrops+by+obliteration+of+the+endolymphatic+duct.">Surgical induction of endolymphatic hydrops by obliteration of the endolymphatic duct.</a> Journal of Visualized Experiments. (35), (2010).</li><li>Lichtenhan, J. T., Cooper, N. P., Guinan, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+auditory+threshold+estimation+technique+for+low+frequencies:+proof+of+concept.">A new auditory threshold estimation technique for low frequencies: proof of concept.</a> Ear and Hearing. 34 (1), 42-51 (2013).</li><li>Lichtenhan, J. T., Hartsock, J., Dornhoffer, J. R., Donovan, K. M., Salt, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+delivery+into+the+cochlear+apex:+Improved+control+to+sequentially+affect+finely+spaced+regions+along+the+entire+length+of+the+cochlear+spiral.">Drug delivery into the cochlear apex: Improved control to sequentially affect finely spaced regions along the entire length of the cochlear spiral.</a> Journal of Neuroscience Methods. 273, 201-209 (2016).</li><li>Lichtenhan, J. T., Hartsock, J. J., Gill, R. M., Guinan, J. J., Salt, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+auditory+nerve+overlapped+waveform+(ANOW)+originates+in+the+cochlear+apex.">The auditory nerve overlapped waveform (ANOW) originates in the cochlear apex.</a> Journal of the Association for Research in Otolaryngology. 15 (3), 395-411 (2014).</li><li>Lichtenhan, J. T., Hirose, K., Buchman, C. A., Duncan, R. K., Salt, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+administration+of+2-Hydroxypropyl-Beta-Cyclodextrin+into+guinea+pig+cochleae:+Effects+on+physiological+and+histological+measurements.">Direct administration of 2-Hydroxypropyl-Beta-Cyclodextrin into guinea pig cochleae: Effects on physiological and histological measurements.</a> PloS One. 12 (4), e0175236 (2017).</li><li>Schneider, C. A., Rasband, W. S., Eliceiri, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIH+Image+to+ImageJ:+25+years+of+image+analysis.">NIH Image to ImageJ: 25 years of image analysis.</a> Nature Methods. 9 (7), 671-675 (2012).</li></ol>

提出的外膜法在实现组织学确认的内溶性水ps和低频听力损失方面成功率为86%。该方法在术后第30天可靠地获得了内溶性水ps组织学证据，这与先前使用内膜方法2的研究一致。该方法对现有方法的意义是不需要 CSF 泄漏，从而消除了一个潜在的混淆变量，该变量已被建议导致内性卷3的补偿性、暂时扩展。总体而言，该方法演示了一种快速、安全、可靠的诱导实验...

内部溶性水液是鳞状介质的扩大。可以使用scala介质的横截面区域测量内性水ps的存在。据认为，临床内心水ps可能与低频感神经性听力损失有关，如梅尼埃病中所看到的。但听力损失的来源仍不清楚。为了充分研究与内溶性水ps相关的低频听力损失的起源，需要一个可靠的模型。
1965年，木村和舒克内赫特描述了如何使用内膜方法1在豚鼠中诱导内淋巴水皮。他们的技术涉及使用后颅口方法访问手术和亚弧线。这些步骤包括切口杜拉，用林格溶液浸湿棉垫收回小脑，以及钻取内心管和内心囊的中间部分。骨蜡然后放入手术管中，将内质导管与远端内溶性囊分离开来。颅内切除术缺陷通过放置可吸收的明胶粉（例如，凝胶泡胶）和重合上覆的肌肉而关闭。在术后的第1、3、7、14、21和30天，不断发现内溶性水ps的组织证据，这表明内核方法是诱导组织确认的内溶性水培的可靠方法。使用与木村和舒克内赫特相同的分道法，但不同的时间点，萨尔特和德莫特证实，在第4天和第2天以后，在耳蜗的第二回合中，scala介质显著扩大。虽然原始研究中没有报告使用木村和舒克内赫特的内膜方法诱导脑脊液（CSF）泄漏的实际发病率，但CSF泄漏的存在可能会增加脑膜炎的风险。有人建议，CSF的丧失可能导致淋巴流出，导致同时暂时扩大几内亚猪3的内部性体积。诱导内淋巴水ps的外膜方法将是一个更安全的选择。
1989年，安德鲁斯和博默描述了两种超前手术方法，通过中颅刀法或后颅骨法方法，达到内质囊和导管，以消灭内心囊4。他们描述了用钻石钻去除手术线，然后钻取内心囊的中间部分，或者使用精细的拾取来破坏内溶性囊和管道。1993年，李、赖特和迈尔霍夫描述了类似的方法，包括钻穿内膜囊和管道，但不同之处在于，它们同时阻塞了耳蜗水渠5。它们表明，在消灭内溶性囊和阻塞耳蜗水渠四周后，通过组学评估的内淋巴水肿的存在。Megerian等人是第一个发表视频文章，展示内膜囊和管道的外层消融，涉及直接钻在手术的中段进入内溶囊和管道6。他们展示了在手术后28周牺牲的豚鼠中内性水蛋白的免疫学，以及在16千赫兹区域6的听力损失。能否在早期时间点用外听觉方法诱导组织学确认的内方水ps和低频听力损失，目前尚不得而知。
本报告的总体目标是演示一种外部方法，通过在术后30天内诱导实验性内膜水肿，通过消灭内溶性囊，用精细的挑料伤害内膜导管。使用这种技术背后的原理是避免在岩石上钻骨，从而消除意外伤害杜拉和导致CSF泄漏的风险，降低后半圆形运河受伤的可能性，并降低对sigmoid正着肠受伤的风险。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>12 mL syringe</td>
			<td>Henke-Sass Wolf</td>
			<td>5100-X00V0</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL and 3 mL syringe</td>
			<td>BD Precision(Ordered from Fischer Sci)</td>
			<td>14-826-87 15859152</td>
			<td></td>
		</tr>
		<tr>
			<td>27.5 butterfly gauge needle</td>
			<td>Terumo Surflo Winged Infusion Set, Terumo Corporation, Japan) (Ordered from McKesson)</td>
			<td>448407</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 suture</td>
			<td>McKesson</td>
			<td>1034507</td>
			<td></td>
		</tr>
		<tr>
			<td>4x4 gauze sponges</td>
			<td>Dukal (Ordered form McKesson)</td>
			<td>374454</td>
			<td></td>
		</tr>
		<tr>
			<td>60mL syringe</td>
			<td>Fisher Sci</td>
			<td>22-031-375</td>
			<td></td>
		</tr>
		<tr>
			<td>Anspach otologic drill</td>
			<td>Anspach</td>
			<td>SC2100</td>
			<td></td>
		</tr>
		<tr>
			<td>atipamezole</td>
			<td>Zoetis</td>
			<td>107204-6</td>
			<td></td>
		</tr>
		<tr>
			<td>autoclave</td>
			<td>Fisher sci</td>
			<td><a href="https://www.fishersci.com/shop/products/fisher-scientific-isotemp-general-purpose-heating-drying-ovens/151030508">15-103-0508</a></td>
			<td></td>
		</tr>
		<tr>
			<td>autoclave bags</td>
			<td>McKesson</td>
			<td>524881</td>
			<td></td>
		</tr>
		<tr>
			<td>bayonet separator</td>
			<td>Olympus</td>
			<td>AL 130564</td>
			<td></td>
		</tr>
		<tr>
			<td>bupivicaine</td>
			<td>auro Medics Pharma</td>
			<td>555150-169-10</td>
			<td></td>
		</tr>
		<tr>
			<td>clear sterile drape</td>
			<td>3M</td>
			<td>1020</td>
			<td></td>
		</tr>
		<tr>
			<td>cotton balls</td>
			<td>Fisherbrand (ordered from Fisher Sci)</td>
			<td>22-456-885</td>
			<td></td>
		</tr>
		<tr>
			<td>cotton swabs</td>
			<td>McKesson</td>
			<td>508716</td>
			<td></td>
		</tr>
		<tr>
			<td>diamond burrs #3, #2, #1, and #0.5 mm</td>
			<td>Anspach</td>
			<td>QD8-3SD; QD8-2SD; QD8-1SD; QD8-05SD</td>
			<td></td>
		</tr>
		<tr>
			<td>diaper pad</td>
			<td>McKesson</td>
			<td>945330</td>
			<td></td>
		</tr>
		<tr>
			<td>disposable 15 blade</td>
			<td>Swann-Morton</td>
			<td>0305</td>
			<td></td>
		</tr>
		<tr>
			<td>enrofloxacin</td>
			<td>Hospira</td>
			<td>0409-4888-01</td>
			<td></td>
		</tr>
		<tr>
			<td>epinephrine</td>
			<td>McKesson</td>
			<td><a href="https://www.findacode.com/code.php?set=NDC&amp;c=63739-0456">63739-0456</a></td>
			<td></td>
		</tr>
		<tr>
			<td>eye ointment</td>
			<td>Dechra Vet Products</td>
			<td>17033-211-38</td>
			<td></td>
		</tr>
		<tr>
			<td>Freer elevator</td>
			<td>Grace Medical</td>
			<td>215100FX</td>
			<td></td>
		</tr>
		<tr>
			<td>gelfoam</td>
			<td>Pfizer (Ordered from McKesson)</td>
			<td>82830</td>
			<td></td>
		</tr>
		<tr>
			<td>hair trimmers</td>
			<td>Oster Power Pro Cordless (ordered from Amazon)</td>
			<td>078400-020-000</td>
			<td></td>
		</tr>
		<tr>
			<td>iodine scrub</td>
			<td>Purdue Pharma (ordered from mcKesson)</td>
			<td>521243</td>
			<td></td>
		</tr>
		<tr>
			<td>iris scissors</td>
			<td>Olympus</td>
			<td>CL-542114</td>
			<td></td>
		</tr>
		<tr>
			<td>ketamine</td>
			<td>Henry Schein Animal Health</td>
			<td>55853</td>
			<td></td>
		</tr>
		<tr>
			<td>lactated ringers</td>
			<td>B. Braun Medical (ordered from McKesson)</td>
			<td>186662</td>
			<td></td>
		</tr>
		<tr>
			<td>lancet knife by Rosen</td>
			<td>Grace Medical</td>
			<td>151100FX</td>
			<td>referred to as curette in the text</td>
		</tr>
		<tr>
			<td>lubricant</td>
			<td>Milex (ordered from Cooper Surgical)</td>
			<td>MX5030</td>
			<td></td>
		</tr>
		<tr>
			<td>masking tape</td>
			<td>3M (ordered from fisher sci</td>
			<td>19047259</td>
			<td></td>
		</tr>
		<tr>
			<td>metal rectangle basin</td>
			<td>Amazon</td>
			<td>B07NQDBC6T</td>
			<td></td>
		</tr>
		<tr>
			<td>needle holder</td>
			<td>Olympus</td>
			<td>CR 213015-ENT</td>
			<td></td>
		</tr>
		<tr>
			<td>needles: 27 gage, 18 gauge</td>
			<td>BD Precision(Ordered from Fischer Sci)</td>
			<td><a href="https://www.fishersci.com/shop/products/bd-general-use-precisionglide-hypodermic-needles-20/1482648">14-826-48 14-826-5D</a></td>
			<td></td>
		</tr>
		<tr>
			<td>neonatal warming isollete</td>
			<td>Air Borne Life Support Systems</td>
			<td>731-1800</td>
			<td></td>
		</tr>
		<tr>
			<td>operating microscope</td>
			<td>Carl Zeiss</td>
			<td>OPMI pico</td>
			<td></td>
		</tr>
		<tr>
			<td>oxygen tank</td>
			<td>AirGas</td>
			<td>OX USP200</td>
			<td></td>
		</tr>
		<tr>
			<td>pulse ox</td>
			<td>CapnoTrue (Ordered from Medacx)</td>
			<td>M-3090112001</td>
			<td></td>
		</tr>
		<tr>
			<td>rectal probe with heating blanket</td>
			<td>Harvard Apparatus</td>
			<td>probe: PY2 50-7217 Heating Blanket: PY2 50-7214</td>
			<td></td>
		</tr>
		<tr>
			<td>red body holder</td>
			<td>Lichtenhan Lab</td>
			<td>N/A</td>
			<td>In-house product</td>
		</tr>
		<tr>
			<td>right angle</td>
			<td>Olympus</td>
			<td>BV-230337</td>
			<td></td>
		</tr>
		<tr>
			<td>rosen needle</td>
			<td>Olympus</td>
			<td>AM-130566</td>
			<td>customized, it is the instrument I use to tear the sac</td>
		</tr>
		<tr>
			<td>rubber tubing for O2 administration</td>
			<td>Fisher Sci</td>
			<td>14-171-104</td>
			<td></td>
		</tr>
		<tr>
			<td>saran wrap</td>
			<td>Fisher Sci</td>
			<td>NC9617977</td>
			<td></td>
		</tr>
		<tr>
			<td>stereotactic head holder</td>
			<td>WUSTL Instrument Machine Shop</td>
			<td>N/A</td>
			<td>In-house product</td>
		</tr>
		<tr>
			<td>sterile drapes</td>
			<td>Cardinal Health</td>
			<td>7553</td>
			<td></td>
		</tr>
		<tr>
			<td>suction tube by Baron</td>
			<td>Grace Medical</td>
			<td>034903FX 034905FX</td>
			<td>#3 and #5 Suction</td>
		</tr>
		<tr>
			<td>tissue forceps adson brown</td>
			<td>Grace Medical</td>
			<td>325112FX</td>
			<td></td>
		</tr>
		<tr>
			<td>Weitlander retractor</td>
			<td>Olympus Grace Medical</td>
			<td>BL200011 100313FX</td>
			<td></td>
		</tr>
		<tr>
			<td>xylazine</td>
			<td>Akorn</td>
			<td>59399-110-20</td>
			<td></td>
		</tr>
	</tbody>
</table>

a revised surgical approach to induce endolymphatic hydrops in the guinea pig

内溶性水ps是一种鳞状体的扩大，通常与梅尼埃的疾病有关，尽管病理生理学机制仍不清楚。为了充分研究内溶性水ps的特性，如低频听力损失的起源，需要一个可靠的模型。豚鼠是一个很好的模型，因为它听到低频区域，这是假定受到内溶性水液的影响。先前的研究表明，内淋巴水肿可以通过内膜或外部方法手术诱发，这些方法涉及在内膜导管和囊上钻孔。然而，是否有可能创建一个内向水肿模型使用外膜方法，避免危险的钻在内膜管道和囊是未知的。这项研究的目的是演示一种经过修订的外膜方法，通过在术后30天内诱导实验性内膜水肿，通过消灭内目囊，用精细的挑料伤害内膜导管。样本大小包括七只豚鼠。对听力进行了功能测量，随后采集了时间骨骼进行组学分析。该方法在实现内性水ps方面成功率为86%。脑脊液泄漏的风险很小。样本中未发生术中半圆形运河的围手术死亡或伤害。该方法演示了一种安全可靠的方法，在相对快速的时间点30天诱导内性水解。临床意义在于，该方法为进一步探索低频听力损失的起源提供了可靠的模型，这种损失可与内溶性水ps相关。

提出的方法采用外部方法，在7头豚鼠中，由2头雄性猪和5只雌性猪组成，用精挑剂来消灭内膜囊，并伤害内心管。手术的平均持续时间从切口到闭合2小时。总钻孔时间从5-10分钟不等。豚鼠需要长达4小时才能完全从麻醉中出来。样本中没有术内或术后死亡。任何豚鼠的后半圆形运河或杜拉没有受伤。一只豚鼠（不包括数据分析），对sigmoid罪子的损伤。
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A Revised Surgical Approach to Induce Endolymphatic Hydrops in the Guinea Pig

本文演示了一种外膜方法，用于消灭豚鼠内膜囊，用精细的挑料伤害内膜导管，以诱导实验性内膜水肿。

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) ...

使用这个程序，内淋巴水培可以相对快速诱导豚鼠与足够的成功率。该技术的主要优点是，内淋巴水培可以诱导与最小的风险CSF泄漏或伤害后半圆运河。在确认对踏板反射缺乏反应后，将12毫升的皮下液体玻利瓦尔的哺乳环器溶液注射到麻醉豚鼠的背面，将豚鼠放在加热垫上的苏皮位置。将 27.5 量的蝴蝶针放在针内，确认正确放置，仅在针尖内存在空气，并放置豚鼠在易发位置。将头部固定到立体定向支架中，将脉冲血小计固定到未发人动的脚上。使用直肠探针和加热毯将体温保持在 38 摄氏度，并在动物的眼睛上涂抹软膏。将一块遮蔽胶带放在背面，以沿皮肤提供足够的张力，将胶带的末端固定到立体定向支架上。然后自由准备皮肤覆盖的奥奇普特和后颈与三个连续的碘磨砂和70%酒精，并放置一个无菌披肩在动物。开始外科手术，使用15号刀片，使一个小中线切口沿后侧切口延伸到后颈。一旦下皮，使用虹膜剪刀将右后颈肌从腹骨分离，用无菌棉球的压力控制任何出血。使用解剖显微镜和数字三，二，和一毫米金刚石毛刺与5-0吸气和无菌灌溉的组合，执行颅骨切除术，边界远至外部腹体峰横向，羔羊脊优越，腹状质缝合线在地中海，和前体巨无霸的背边低。并使用 0.5 毫米金刚石毛刺对西格莫德正弦进行骨架化。轻轻地将一小块盐水湿润的棉花放在骨头下面，同时将腹骨与杜拉分离。小心地取出覆盖鼻窦的骨头，并使用棉球轻轻收回西莫德鼻窦。小心地收回西格莫德正弦，将操作器识别为位于 petrous 时间骨内的狭缝状结构。内淋巴囊的外侧部分可以可视化为操作体和覆盖西莫德正窦的杜拉状连接。对西格莫德正弦应用温和的中缩，以清楚地可视化内淋巴囊的外质部分，并增加囊的外间和内分部之间的张力。使用细角拾取，轻轻清除内淋巴囊的中间部分，并沿骨内侧广泛刮取操作体内的细取。对于此过程的成功，杜拉和操作器之间没有可见的连接至关重要。使用小花栗沿着时间骨刮，以获得骨片，并慷慨地用骨片包装手术。用骨蜡密封手术库，注意没有多余的蜡脱落到头骨，并使用骨蜡来掩盖颅骨切除术的缺陷。然后以中断的方式用 4-0 编织可吸收缝合线近似后颈椎肌肉，并使用 4-0 编织可吸收缝合线近似深层和表皮层，以 4-0 编织可吸收缝合线近似深层。在这个具有代表性的实验中，对时间骨骼的组织学分析揭示了整个右耳蜗的内淋巴水合，而左耳蜗则有七分之六。每个转弯时对斯卡拉介质的横截面面积进行量化表明，与在控制动物中观察到的相比，在消灭内淋巴囊30天后存活30天的动物的耳朵中，横截面区域一般较大。在具有内淋巴水培的动物中，听觉神经重叠波形阈值增加，表明存在低频听力损失，而大多数高频耳蜗复合作用潜在阈值在8至20千赫之间。在正常范围内。如果 dura 和 operculum 之间的区域在清除囊后干燥，请尝试在西格莫德正弦上放置额外的缩回，以确保它和蛋白石之间没有明显的连接。按照这个程序，可以进行关于耳蜗解剖和生理学的其他研究，以解决有关内淋巴水合的重要研究问题。

Research

JoVE Journal

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科研

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