我们承认日本促进科学协会 (KAKENHI 赠款至 [26893261])、Okinaka 医学研究纪念研究所 (对 KN) 以及日本卫生部、劳工和福利部 (TN) 提供的资助;Mrs. Kohichi 蒲田和 Atsushi 山在崎玉医科大学国际医学中心的病理学系, 为其组织化学和免疫组织化学染色提供了良好的帮助;以及 Dr. 塞尔索·阿莫林 e. 戈麦斯-桑切斯提供的小鼠单克隆 CYP11B2 抗体和大鼠单克隆 CYP11B1 抗体。

这项研究由崎玉医科大学国际医疗中心和横滨 Rosai 医院的机构审查委员会批准 (批准号: 16-093 和 26-38, 分别)。在手术前, 所有患者均获得书面同意. 
 1. 患者准备 <ol> <li> 在检查床上设置病人. </li>
	<li> 在手术过程中, 在上臂 (或左腿) 上放置一条静脉导管, 用于用药. </li>
	<li> 在适当的皮肤消毒和局部麻醉后, 将通路鞘插入右股静脉. </li>
	<li> 收集血液样本 (1 毫升) 从右股静脉 (外周血样本的管理前促). </li>
</ol> 2。导尿管 <ol> <li> ssAVS 右肾上腺电视 <ol> 插入任一导管与 mk 肾上腺-R 形状 (<li> 图 1A 和 1B ) 或导管与 mk X 形状 ( 图 1C )进入右肾上腺静脉 (雷夫), 如先前所报告的 15 , 16 , 收集血液样本 (1 毫升), 并取出导管 (右骑士样本前的管理促)。如果需要, 使用微导管. </li>
		<li> 为右肾上腺选择合适的父导管: 当和 #34; 下腔和 #34 的长径; 短于25毫米, 使用 MK 肾上腺-R。否则, 使用 MK x. </li>
		如果需要, <li> 重新定型父导管。
		<ol> <li> 基于 CT 图像, 塑造选定导管的宽度 ( 图 1A 或 1C 中的红色双向箭头) 以适合 #34; 下腔和 #34 的长直径; 15 (也请参阅案例 #1)和导管尖端的角度, 以适应和 #34; 修改了雷夫和 #34 的横向角度; 15 (也请参见案例 #1). </li>
			<li> 重新塑造导管, 同时将高温蒸汽从开水 (如电子水壶) 应用到导管。在其他地方 15 中描述了详细的过程. </li>
		</ol> </li> <li> 将父导管的尖端插入雷夫, 并以适当的角度 (雷夫的平均修改横向角度: 123.6 和 #176;) 15 和深度 (1-2 mm). 
		注: 准确的母导管放置是将微导管送至靶电视的关键. </li>
		<li> 操作母导管以改变导管尖端的方向, 以便瞄准其中一个正确的电视, 然后插入一个含盐水的微导管, 丝. 
		注: 推拉母导管会改变母导管的垂直角度, 即 当 3 d 型导管向脚部拉动时, 导管尖端向上, 而导管尖端则向下定向, 当导管被推挤 (参见案件 #1 下面)。一旦导管尖端插入雷夫出口的适当角度和深度, 微导管可以插入。这是重要的考官, 以防止病人的呼吸深度或垂直角度可能改变。上面所述的微导管插入丝到电视中. </li>
		<li> 用数字减影血管造影通过微导管进行造影, 少量生理盐水稀释 (1:1) 造影剂 (0.1-0.3 毫升; 普罗注射液), 轻轻冲洗。在血浆醛固酮浓度 (PAC) 和血浆皮质醇浓度 (PCC) 的测量中, 缓慢收集1毫升血液样本. </li>
		<li> 微导管稍稍向后拉, 用至少0.5 毫升盐水冲洗微导管线。在下一台电视上瞄准母导管提示. </li>
		<li> 对所有正确的电视重复步骤2.1.6 和 2.1.7. </li>
	</ol> </li> <li> ssAVS 左肾上腺电视 <ol> <li> 将 L 形导管 ( 图 2 ) 插入左肾上腺静脉 (LAV), 收集血液样本 (1 毫升), 并取出导管 (左骑士样本前管理促)。使用微导管. </li>
		<li> 放置父导管. 
		注意: 解剖上, 左肾上腺中央静脉与下膈静脉汇合, 静脉血流到 LAV 8 。因此, 要只收集左肾上腺组织的静脉血标本, 需要插入左肾上腺中央静脉, 即 , 在与膈下静脉合并前插入穴位, 这通常需要微导管, 甚至骑士。当进行肾上腺中央静脉造影, 类似于右侧, 这是重要的, 以确定左肾上腺电视, 特别是侧电视 (见和 #34; 案例 #2 作为一个例子, LAV-ssAVS 和 #34;)。使用 L 型导管的首选。部分 #1, #2, 和 #3 ( 图 2 ) 的导管适合下腔静脉, 肾静脉, 以及 LAV 的共同躯干, 从而允许部分 #3 稳定地坐在共同的躯干. </li>
		<li> 将微导管插入左侧肾上腺电视, 并从左电视收集血液样本。查看步骤 2.1.5-2.1.6 以上为一般程序的右肾上腺电视. 
		注: 微导管和丝 (与右 ssAVS 相同) 插入电视 (通常是上级-中位, 上侧和侧电视), 如下所述 #2 的情况下, 并收集血液样本. </li>
	</ol> </li> </ol> 3。注射后 <ol> <li> 注入200和 #181; 通过静脉线进行合成肾上腺皮质激素激素 (促、丸) g, 然后连续促50和 #181; g/分钟 </li>
	<li> 十五分钟后, 促注射, 再次执行骑士, 如上文所述, 和 ssAVS, 如步骤2.1 和2.2。收集1毫升血液. </li>
	<li> 从右股静脉采集血液样本 </li>
	<li> 卸下所有导管和通道鞘, 完成便秘后的骑士和 ssAVS 检查. 
	注: 有关一般肾上腺静脉取样技术的详细资料 (除3.2 的步骤外), 请参阅其他教科书或期刊 17 . </li>
</ol>

<ol>
	<li>Funder, J. W., 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+Management+of+Primary+Aldosteronism:+Case+Detection,+Diagnosis,+and+Treatment:+An+Endocrine+Society+Clinical+Practice+Guideline.">The Management of Primary Aldosteronism: Case Detection, Diagnosis, and Treatment: An Endocrine Society Clinical Practice Guideline.</a> J Clin Endocrinol Metab. 101 (5), 1889-1916 (2016).</li><li>Akehi, 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=Proposed+diagnostic+criteria+for+subclinical+Cushing's+syndrome+associated+with+adrenal+incidentaloma.">Proposed diagnostic criteria for subclinical Cushing's syndrome associated with adrenal incidentaloma.</a> Endocr J. 60 (7), 903-912 (2013).</li><li>Choi, 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=K++channel+mutations+in+adrenal+aldosterone-producing+adenomas+and+hereditary+hypertension.">K+ channel mutations in adrenal aldosterone-producing adenomas and hereditary hypertension.</a> Science. 331 (6018), 768-772 (2011).</li><li>Beuschlein, 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=Somatic+mutations+in+ATP1A1+and+ATP2B3+lead+to+aldosterone-producing+adenomas+and+secondary+hypertension.">Somatic mutations in ATP1A1 and ATP2B3 lead to aldosterone-producing adenomas and secondary hypertension.</a> Nat Genet. 45 (4), 440-444 (2013).</li><li>Scholl, U. I., 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=Somatic+and+germline+CACNA1D+calcium+channel+mutations+in+aldosterone-producing+adenomas+and+primary+aldosteronism.">Somatic and germline CACNA1D calcium channel mutations in aldosterone-producing adenomas and primary aldosteronism.</a> Nat Genet. 45 (9), 1050-1054 (2013).</li><li>Azizan, E. 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=Somatic+mutations+in+ATP1A1+and+CACNA1D+underlie+a+common+subtype+of+adrenal+hypertension.">Somatic mutations in ATP1A1 and CACNA1D underlie a common subtype of adrenal hypertension.</a> Nat Genet. 45 (9), 1055-1060 (2013).</li><li>Tamura, 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=Somatic+KCNJ5+mutation+occurring+early+in+adrenal+development+may+cause+a+novel+form+of+juvenile+primary+aldosteronism.">Somatic KCNJ5 mutation occurring early in adrenal development may cause a novel form of juvenile primary aldosteronism.</a> Mol Cell Endocrinol. 441, 134-139 (2017).</li><li>McLachlan, M. S., Roberts, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demonstration+of+the+normal+adrenal+gland+by+venography+and+gas+insufflation.">Demonstration of the normal adrenal gland by venography and gas insufflation.</a> Br J Radiol. 44 (525), 664-671 (1971).</li><li>Nishikawa, T., Matsuzawa, Y., Saito, J., Omura, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+it+Possible+to+Extirpate+Cardiovascular+Events+in+Primary+Aldosteronism+After+Surgical+Treatment.">Is it Possible to Extirpate Cardiovascular Events in Primary Aldosteronism After Surgical Treatment.</a> Jpn Clin Med. 1, 21-23 (2010).</li><li>Nishikawa, T., Matsuzawa, Y., Saito, J., Omura, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Super-selective+ACTH-stimulated+adrenal+venous+sampling+can+simply+differentiated+bilateral+adrenal+hyperplasia+from+bilateral+adenomas+in+primary+aldosteronism.">Super-selective ACTH-stimulated adrenal venous sampling can simply differentiated bilateral adrenal hyperplasia from bilateral adenomas in primary aldosteronism.</a> , 35-36 (2009).</li><li>Nishimoto, 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=Case+Report:+Nodule+Development+From+Subcapsular+Aldosterone-Producing+Cell+Clusters+Causes+Hyperaldosteronism.">Case Report: Nodule Development From Subcapsular Aldosterone-Producing Cell Clusters Causes Hyperaldosteronism.</a> J Clin Endocrinol Metab. 101 (1), 6-9 (2016).</li><li>Miekos, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomical+basis+of+radiodiagnosis+of+the+adrenal+gland.">Anatomical basis of radiodiagnosis of the adrenal gland.</a> Int Urol Nephrol. 11 (3), 193-200 (1979).</li><li>Omura, M., Saito, J., Matsuzawa, Y., Nishikawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supper-selective+ACTH-stimulated+adrenal+vein+sampling+is+necessary+for+detecting+precisely+functional+state+of+various+lesions+in+unilateral+and+bilateral+adrenal+disorders,+inducing+primary+aldosteronism+with+subclinical+Cushing's+syndrome.">Supper-selective ACTH-stimulated adrenal vein sampling is necessary for detecting precisely functional state of various lesions in unilateral and bilateral adrenal disorders, inducing primary aldosteronism with subclinical Cushing's syndrome.</a> Endocr J. 58 (10), 919-920 (2011).</li><li>Nishikawa, T., Omura, M., Saito, J., Matsuzawa, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primary+aldosteronism:+comparison+between+guidelines+of+the+Japanese+and+the+US+Endocrine+Society.">Primary aldosteronism: comparison between guidelines of the Japanese and the US Endocrine Society.</a> Expert Rev. Endocrinol. Metab. 7 (6), 637-645 (2012).</li><li>Araki, T., Okada, H., Onishi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+catheter+shape+influence+the+success+of+right+adrenal+venous+sampling?+The+interaction+of+catheter+shape+to+anatomical+factors+on+CT.">Does catheter shape influence the success of right adrenal venous sampling? The interaction of catheter shape to anatomical factors on CT.</a> Jpn J Radiol. 34 (11), 707-717 (2016).</li><li>Young, W. F., Stanson, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+are+the+keys+to+successful+adrenal+venous+sampling+(AVS)+in+patients+with+primary+aldosteronism?.">What are the keys to successful adrenal venous sampling (AVS) in patients with primary aldosteronism?.</a> Clin Endocrinol (Oxf). 70 (1), 14-17 (2009).</li><li>Harsha, A., Trerotola, S. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+aspects+of+adrenal+vein+sampling.">Technical aspects of adrenal vein sampling.</a> J Vasc Interv Radiol. 26 (2), 239 (2015).</li><li>Nishikawa, T., 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=Guidelines+for+the+diagnosis+and+treatment+of+primary+aldosteronism--the+Japan+Endocrine+Society+2009.">Guidelines for the diagnosis and treatment of primary aldosteronism--the Japan Endocrine Society 2009.</a> Endocr J. 58 (9), 711-721 (2011).</li><li>Nishimoto, 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=Immunohistochemistry+of+aldosterone+synthase+leads+the+way+to+the+pathogenesis+of+primary+aldosteronism.">Immunohistochemistry of aldosterone synthase leads the way to the pathogenesis of primary aldosteronism.</a> Mol Cell Endocrinol. , (2016).</li><li>Nishimoto, 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=Adrenocortical+zonation+in+humans+under+normal+and+pathological+conditions.">Adrenocortical zonation in humans under normal and pathological conditions.</a> J Clin Endocrinol Metab. 95 (5), 2296-2305 (2010).</li><li>Funder, J. W., 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=Case+detection,+diagnosis,+and+treatment+of+patients+with+primary+aldosteronism:+an+endocrine+society+clinical+practice+guideline.">Case detection, diagnosis, and treatment of patients with primary aldosteronism: an endocrine society clinical practice guideline.</a> J Clin Endocrinol Metab. 93 (9), 3266-3281 (2008).</li><li>Heitzer, E., Auer, M., Ulz, P., Geigl, J. B., Speicher, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+tumor+cells+and+DNA+as+liquid+biopsies.">Circulating tumor cells and DNA as liquid biopsies.</a> Genome Med. 5 (8), 73 (2013).</li><li>Alix-Panabieres, C., Pantel, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+Applications+of+Circulating+Tumor+Cells+and+Circulating+Tumor+DNA+as+Liquid+Biopsy.">Clinical Applications of Circulating Tumor Cells and Circulating Tumor DNA as Liquid Biopsy.</a> Cancer Discov. 6 (5), 479-491 (2016).</li><li>Pinzani, P., 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=Detection+of+circulating+tumor+cells+in+patients+with+adrenocortical+carcinoma:+a+monocentric+preliminary+study.">Detection of circulating tumor cells in patients with adrenocortical carcinoma: a monocentric preliminary study.</a> J Clin Endocrinol Metab. 98 (9), 3731-3738 (2013).</li><li>Pantel, 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=Circulating+epithelial+cells+in+patients+with+benign+colon+diseases.">Circulating epithelial cells in patients with benign colon diseases.</a> Clin Chem. 58 (5), 936-940 (2012).</li><li>Chiu, L. 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=Identification+of+differentially+expressed+microRNAs+in+human+hepatocellular+adenoma+associated+with+type+I+glycogen+storage+disease:+a+potential+utility+as+biomarkers.">Identification of differentially expressed microRNAs in human hepatocellular adenoma associated with type I glycogen storage disease: a potential utility as biomarkers.</a> J Gastroenterol. 49 (8), 1274-1284 (2014).</li><li>Nishimoto, 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=Aldosterone-stimulating+somatic+gene+mutations+are+common+in+normal+adrenal+glands.">Aldosterone-stimulating somatic gene mutations are common in normal adrenal glands.</a> Proc Natl Acad Sci U S A. 112 (33), E4591-E4599 (2015).</li></ol>

作者没有什么可透露的。

本文介绍了具有代表性的案例结果的 ssAVS 技术。病例 #1-2 例, 病例 #3 分别根据 ssAVS 结果进行手术治疗。此外, #1-3 病例的结果表明, ssAVS 样本中的类固醇激素浓度清楚地反映了上游肾上腺组织, 特别是肿瘤的激素活性, 推测是因为肿瘤血流是直接得到的。ssAVS 方法可能在鉴别肾上腺皮质病变 (例如, #1-2), 在明确诊断 IHA (如:如, 病例 #3) 和基础科学研究中发挥不可估量的作用, 以阐明pathophysiologies 肾上腺皮质疾病, 并发现新的生物标志物的这些疾病如下所述。
pa1、1821建议执行骑士通过计算侧比率 ([高 pac/pcc]/[低 pac/pcc]) 来鉴别 pa 的单侧肾上腺损伤。然而, 这种计算可能导致误诊时, 皮质醇产生的病变共存。可能的皮质醇产生腺瘤的情况下 #2 清楚地产生过量的皮质醇在右肾上腺, 和皮质醇生产左肾上腺被压制。重要的是要注意, 许多 apa 也产生过量的皮质醇, 因为他们经常表达的 CYP11B2 和 CYP11B120。骑士侧比率的另一个限制是, 它不能区分双边 apa 与特发性症 (例如, 案例 #3)。
此外, ssAVS 血液样本可能会大大促进激素过剩疾病的研究和治疗的进展。例如, 从腺瘤收集的 ssAVS 血液目录可能含有高浓度的循环肿瘤细胞及其 DNA22,23。以往的研究报告, 液体活检方法是临床有用的鉴别肾上腺皮质癌从腺瘤24和诊断良性腺瘤疾病25,26。为了证明这一概念, 目前正在尝试检测 APA 相关的突变, 包括使用 ssAVS 样本的KCNJ5和高性能的下一代排序器71119 ,27, 这可能有助于将来的 APA 治疗。液态活检细胞也可能为研究人员提供在 IHA 上进行分子分析的机会, 这是目前不可能的, 因为这种疾病不能通过手术治疗。除了液体活检方法外, 单纯的肿瘤流出样可能有助于代谢研究, 以识别新的类固醇生物标志物。
ssAVS 方法的关键技术步骤是: (i) 在中央肾上腺静脉造影中识别各支流静脉。(ii) 在静脉造影时, 使用少量造影剂 (0.1-0.3 毫升), 轻轻冲洗, 避免肾上腺出血。(iii) 轻轻地推进丝, 不需要太多的力, 以避免穿透支流静脉。(iv) 当肾上腺出血发生或怀疑时立即终止该方法。
选择父母导管对 ssAVS 的成功也是至关重要的。关于骑士的雷夫, 荒 et al. 最近报告了一个三维 (3 d) 型导管的用处, 3D 形状的15。带有 R 形 (图 1A和1B) 的导管和带有 X 形 (图 1C) 的导管可作为 ssAVS 3 d 型导管使用。荒 et al. 分析了一些解剖参数的基础上的 CT 表现, 关于成功率的 3 d 型导管15。在单变量分析中 (i) 较短的 "下腔静脉短径 (下)" (ii) 较大的 "长径比下腔静脉的短直径" (iii) 更小的 "雷夫的横向角度" (iv) 更小的 "修改过的横向角度雷夫", 和 (五) 较小的 "雷夫的垂直角与雷夫的成功率相关。在多变量分析中, 只有 (iv) 较小的 "改良横角雷夫" 是成功雷夫插管的独立预测因子。他们的结论是, 多变量分析的结果可能是由于在下腔静脉导管的稳定性;即, 3 d 型导管的宽度 (图 1A和1C中的红色双向箭头) 非常适合 "下腔静脉的长径", 从而稳定了这些导管。总的来说, 当 "下腔静脉的长径" 短于25毫米时, 使用 mk 肾上腺-R, 否则 mk x。
总的来说, ssAVS 的意义是: (一) 它对促进激素过剩疾病研究的贡献 (ii) 通过将激素产生性病变隔离到肾上腺段的水平 (见 #1) (iii.)通过收集纯肾上腺流出来促进对皮质醇过量的评估 (见案例 #2) (iv) 检测实际特发性症 (见案例 #3) (五) 它对推动新疗法的发展作出了贡献, 其中可能包括肾上腺的跨静脉节段消融术。因此, 如果按照本文所述, 任何 angiographer 可能成功地执行 ssAVS 协议, 除了骑士, 并有助于促进研究和治疗肾上腺激素过剩疾病。
ssAVS 与骑士经常在横滨 Rosai 医院和崎玉医学院为 PA 病人。在2014年10月和2015年9月之间, 两个 angiographers (公里和 SM) 对125例 (分别为78和47例) 进行了 ssAVS, 100% 的成功率和合理时间内 (58-130 分钟) 不需要手术的肾上腺破裂或血栓形成。在这种方法中, 一个额外的成本是发生的微导管 (10 倍的费用在日本比传统的导管) 和 PAC/PCC 测量的支流样品。然而, 考虑到该程序的临床和科学效益, 额外的费用是合理的, 至少在工业化国家。ssAVS 和双边肾上腺的一个边际限制是, 它可能无法治愈双边病变, 需要进行后续评估, 包括评估 PA 复发 (见案例 #1)。然而, 对单方面的 PA 案件也是如此。因此, 这些结果表明, 任何 angiographer 都能够按照本视频文章中提供的协议执行 ssAVS 的高成功率。为了促进ssAVS 方法在世界各地, 在横滨 Rosai 医院和玉崎医科大学始终提供实践培训。如果您对此方法感兴趣, 请随时与这些机构联系。

原发性醛 (PA) 和亚临床库欣氏综合征 (SCS) 的情况下, 肾上腺自主产生过量的醛固酮和皮质醇, 分别。在成人中, PA 主要由醛固酮产生腺瘤 (APA) 或特发性症 (IHA)1引起, 而 SCS 主要是由皮质醇产生的腺瘤2引起的。离子通道/泵基因中的体细胞突变, 包括钾通道、内整流亚科 J、成员 5 (KCNJ5), 已在 apa 中被识别, 并与自主醛固酮生成有关3,4,5,6. 家族性症类型 1-3 是 PA 的稀有类型, 而类型3是由KCNJ5生殖细胞变异3引起的。一个新类型的少年 PA 的情况下, 这大概是由于遗传嵌的正常和KCNJ5突变的肾上腺皮质细胞, 最近被确定为7。在少年 PA 的情况下, 肾上腺皮质由正常部分和增生性醛固酮产生的损害, 与正常部分被识别的小说 ' 选择性 ' 肾上腺静脉取样 (ssAVS, 也称为节段性肾上腺静脉取样) 方法的描述。ssAVS 方法允许这种双边 PA 患者手术治疗的双边肾上腺, 同时保留正常部分7。
肾上腺静脉取样最初报告在 1971年8时, 计算机断层扫描 (CT) 尚未开发。在 "常规" 肾上腺静脉取样 (骑士), 导管插入两个肾上腺中央静脉, 血液样本收集。因此, 骑士只适用于判断 pa 的侧, 而不用于确定双边 pa 的手术选择。Kohzoh 牧 (本文作者) 在横滨 Rosai 医院开发了 ssAVS 方法, 以调查 PA7、8、9的其他外科治疗选项, 10,11, 这对 SCS 也很有用, 如情况 #2 (请参见代表性结果) 中所述。
在 ssAVS, 血液样本收集的小肾上腺支流静脉 (电视: 较小的上游分支肾上腺中央静脉) 使用专门的分裂尖端微导管9。右侧和左侧肾上腺通常有三条支流静脉, 即在右侧肾上腺中的上级、侧面和下位电视以及左侧肾上腺中的中位、上侧和侧电视。这些电视12之间的连接很少被识别。因此, ssAVS 能够识别一个特定的肾上腺段 (s) 不产生醛固酮在一个受影响的肾上腺, 从而允许双侧肾上腺手术在双边 PA, 同时保留不影响肾上腺段 (s)。此外, ssAVS 方法也适用于少年 pa 等新型 pa 的病理生理学研究 (如上述7) 和双侧 APA13、14。由于对双边 PAs 的治疗和对其发病机制的说明至关重要, 其中可能包括 "特发性" 症, 因此建议在全世界范围内 ssAVS, 因此, 本文提供了有关方法的详细描述。

<table><tbody><tr><td>CX-Catheter EII with the MK Adrenal-R shape</td><td>Silux co.</td><td>GA-E5F-MK1-60S</td><td>3D catheter for right adrenal venous sampling</td></tr><tr><td>CX-Catheter EII with the MK X shape</td><td>Silux co.</td><td>GA-E5F [MK-X] 60S</td><td>3D catheter for right adrenal venous sampling</td></tr><tr><td>Aqua V3 guidewire</td><td>Formec co.</td><td>HS8616H</td><td>guidewire for the micro-catheter</td></tr><tr><td>Gold Crest Micro-Catheter</td><td>Goldcrest Medic Inc.</td><td>KCV29S1S-OM</td><td>micro-catheter for superselective adrenal venous sampling</td></tr><tr><td>CX catheter-UII with the MK ADRENAL-L shape</td><td>Silux co.</td><td>GA-US5F [MK-3] B65S</td><td>3D catheter for left adrenal venous sampling</td></tr></tbody></table>

a novel method: super-selective adrenal venous sampling

原发性醛 (PA) 和亚临床库欣氏综合征 (SCS) 的情况下, 肾上腺自主产生过量的醛固酮和皮质醇, 分别。传统的肾上腺静脉取样 (骑士) 方法收集来自肾上腺中心静脉的血样, 并可用于鉴别单侧病变 (s) 中过剩激素的侧, 如 PA 病例所示。在骑士队, 血浆皮质醇浓度 (PCCs) 被用来正常化血浆醛固酮浓度 (PACs)。一种新的 "选择性" 肾上腺静脉取样 (ssAVS) 的方法是利用微导管, 收集血液样本从肾上腺支流静脉 (电视)。ssAVS 样本中的 PACs 不需要 PCC 正常化, 因为样本含有有限数量的全身静脉血, 如果有的话。ssAVS 方法可以在两个肾上腺中检测到节段性病变, 这可能由双侧肾上腺, 从而保留无损伤段 (s)。右和左肾上腺通常有三电视,即, 右肾上腺的上位、侧、下位电视以及左侧肾上腺中的中位、上侧和侧电视。在 ssAVS 方法中, 需要特定的母导管和处理它们的技术, 并在此描述。此外, 三例 PA 的 ssAVS 结果提出: 双侧醛固酮产生腺瘤 (apa) (病例 #1), 左 APA 和右侧可能产生的皮质醇分泌腺瘤 (病例 #2), 和特发性症, 其中双边肾上腺段产生过量的醛固酮 (病例 #3)。ssAVS 方法是不难的专家 angiographers, 因此, 建议在全球范围内治疗 PA 的情况下, 骑士不代表一个可行的外科治疗选项。

案例 #1 (YRPA #3472)
案例 #1 是一个 46 year-old 的女性。当她33岁的时候, 她在当地医院住院治疗严重高血压合并低钾血症 (血清钾 1.8 [正常范围: 3.5-5.9] 当量/升)。她被诊断为根据血液测试结果 (PAC 320 [35.7-240] pg/毫升, PRA 和 #60; 0.1 [0.3-2.9] ng/毫升/小时)。CT 显示双侧肾上腺皮质腺瘤 (数据不可用)。在骑士之后 (数据不可用), 她接受了左肾上腺, 但她的 PA 坚持了。第一次手术后十三年 (46 岁), 她被称为横滨 Rosai 医院评估 PA。身体检查是正常的, 除了高身体质量索引 (29.3 [和 #60; 25] 公斤或 m2)。实验室测试是正常的, 除了非常高的 PAC (1490 pg/毫升) 和非常低血清 K (2.2 当量/升)。她的 PAC 是非常高 (2550 [截止: 和 #60; 60] pg/毫升), 甚至4小时后, 2 升盐水 (盐水输液测试), 表明她有严重的 PA。腹部 CT 扫描显示22毫米右侧肾上腺肿瘤 (图 3A)。
骑士和 ssAVS 在刺激下进行了合成肾上腺皮质激素激素 (ACTH)。基于 CT "下腔静脉的长径" (图 3A中红色虚线的长度) 和 "修改过的雷夫的横向角度" (图 3A中红色和蓝色虚线之间的较大角度) 分别为 28 mm 和145度。导管的 "宽度" 和 "尖角" 与 X 形状 (图 1C) 前重塑以适合下腔静脉和雷夫, 详见15。插管进入雷夫的出口很快就完成了, 没有任何困难。右肾上腺静脉造影显示, 侧电视明显扩大 (粉红色箭头在图 3B) 在其分支机构概述了肿瘤的形状, 表明大量的血液从腺瘤流出到这台电视机里。在证实其静脉造影 (图 3C) 后, 插入了一根微导管, 并从侧电视采集血液样本。通过拉动和推导管, 微导管很容易插入到上、下位电视中进行静脉造影 (图 3D和3E), 随后进行取样。
在中央静脉和侧电视的 PAC 是非常高的 (42.2万 pg/毫升和 58.8万 pg/毫升, 分别; 正常范围和 #60; 1.4万 pg/毫升为两个18), 建议肿瘤是一个 APA (图 3C, 表 1)。在高级和下位电视的 PAC 分别为 8230 pg/毫升和 12600 pg/毫升 (图 3D和3E), 这表明这些电视是从正常的肾上腺组织中采集血液的。在中央静脉, 高级电视, 侧电视和下位电视的 PCC 水平相似 (1110 µg/dl, 1150 µg/dl, 1050 µg/dl, 和1080µg/dl, 分别), 表明皮质醇的生产是均匀的整个肾上腺皮质包括肿瘤轴承部分。因此, 通常用于骑士数据分析的 pac/PCC 值与骑士和 ssAVS 的 pac 值是一致的。她接受了部分肾上腺的正常部分。病理检查发现肾上腺皮质腺瘤 (t 在图 3F), 表达醛固酮合成酶 (CYP11B2) 在许多细胞 (t 在图 3G) 和类固醇11β羟化酶 (CYP11B1, 皮质醇合成酵素) 在少量的单元格 (T 在图 3H中), 确认了 APA19、20的诊断。在毗邻正常肾上腺, 虽然 CYP11B2 没有表达的透明球状, 这可能是由于低循环肾素, CYP11B1 表达的透明束和透明网状 (N 在图 3H), 表明皮质醇生产正常。这些病理结果的 APA 和抑制 CYP11B2 表达在相邻的正常肾上腺组织是一致的 ssAVS 结果 (表 1)。手术后, 她的血压 (114/62 柱), 以及 PAC 和 PRA 在她的外周血 (52 pg/毫升和 0.8 ng/毫升/小时, 分别) 正常化没有任何降压药物。
案例 #2 (YRPA #4119)
案件 #2 是 59 year-old 男性与高血压, 因为他是45岁。CT 在例行体检中偶然发现了双侧肾上腺结节, 这是增强的对比培养基 (图 4A)。他被转介到横滨 Rosai 医院, 进一步评估高血压和肾上腺结节。体检正常, 没有明显的 Cushingoid 特征。血液测试正常, 包括 PCC (7.6 [6.2-18.0] µg/dL), ACTH (20.8 [7.2-63.3] pg/毫升), 和 PAC (201 pg/毫升), 除了 PRA (和 #60; 0.2 ng/毫升/小时) 和血清钾 (3.0 当量/升)。盐水灌注试验显示高 PAC (374 pg/毫升)。PCC 在下午11点和在隔夜管理以后1毫克地塞米松是6.8 和 7.2 (切断: 5 和≤1.8) µg/dL, 分别。因此, 他被诊断为 PA 与 SCS2,18。
为了确定哪个肿瘤是导致过量激素分泌的原因, 骑士与 ssAVS 是在合成促肾上腺皮质激素刺激下进行的。左肾上腺静脉造影使用微导管通过导管与 L 形 (图 2) 确定了典型的高级中位数 (图 4B中的黄色箭头)、上级侧 (红色箭头) 和侧电视 (粉红色箭头)。中位电视有一个短的装填瑕疵, 大概归结于腺瘤 (绿色箭头在图 4B)。值得注意的是, 微导管的头部 (图 4B中的黑色箭头) 被放置在肾上腺中央静脉内, 然后与膈下静脉合并, 从而使侧电视的成像通畅。在丝, 微导管插入到上级中位 (静脉造影是不可用的) 和上级侧 (图 4C) 电视的静脉造影和样本收集。横向电视与垂直合并了中央静脉, 这是一个典型的发现。微导管及其丝的尖端被弯曲并插入侧电视, 采集血液样本。如 #1 的情况所述, 骑士从雷夫和 ssAVS 从右上电视 (红色箭头在图 4E), 侧面电视 (粉红色箭头) 的下游肿瘤, 和劣质电视 (黄色箭头) 也执行。
在骑士和 ssAVS 的数据分析中, 使用了 pac 和 pcc 值, 而不是 pac/pcc 值, 因为 pcc 的值在右边和左边的电视中明显变化 (中值和分范围: 分别为99.6 和 70.3-577.5 µg/dL),表 1)。PAC 在左肾上腺中央静脉高 (94800, #60; 1.4万] pg/毫升), 并在左上-中位电视是非常高的 (30.4万 pg/毫升: 3.2 倍, 在中央静脉), 表明左肾上腺肿瘤是损害 PA 负责。然而, PAC 在左上侧和侧面电视是低的 (分别2060和 2240 pg/毫升), 建议他们收集血液从肿瘤部分。左中央静脉的 PCC 上位电视、上侧电视和侧电视 (74.7 µg/dl、87.1 µg/dl、75.7 µg/dl、54.1 µg/dl 分别) 明显低于 #1, 表明皮质醇生产在整个左肾上腺皮质包括肿瘤由于过量的皮质醇生产从右肾上腺, 如下所述。关于右肾上腺 PAC 在右中央静脉 (5190 pg/毫升,即, 在正常范围内和 #60; 1.4万) 和侧电视 (5300 pg/毫升) 高于那些在高级和劣质电视 (1710 和 2180 pg/毫升, 分别),建议右瘤产生少量醛固酮。右侧电视 (1050 µg/dl) 的 PCC 明显高于右侧电视 (112 µg/dl)、右下电视 (420 µg/dl) 和左电视, 这表明右肾上腺肿瘤产生过量的皮质醇 (即皮质醇产生腺瘤) 和引起的 SCS。为了治疗 PA, 患者接受了正确的部分肾上腺, 正常化他的高血压 (136/82 柱没有药) 和 PAC (50 pg/毫升) 手术后3天。病理检查确定了一个肾上腺皮质腺瘤 (t 在图 4F), 表示 CYP11B2 (t 在图 4G), 但不是 CYP11B1 (t 在图 4H), 确认诊断 APA。相邻的正常肾上腺没有表达 CYP11B1 (N 在图 4H), 这表明皮质醇生产被抑制, 由于可能的皮质醇产生腺瘤在相反的一侧。这些病理结果的腺瘤和相邻的肾上腺组织是一致的 ssAVS 的结果 (表 1)。SCS 目前没有治疗, 因为它没有引起高血压或糖耐量受损2。
总的来说, 在 #1 和 #2 的情况下, ssAVS 方法清楚地显示出节段肾上腺激素的产生, 不仅对醛固酮, 而且对皮质醇, 并使这些患者的手术治疗。
案例 #3 (YRPA #8243)
病例 #3 是一名50岁的 year-old 女性, 自48岁以来因严重高血压而头晕。一个高 PAC 到 PRA 比率 ([131 pg/毫升]/[0.3 ng/毫升/小时] = 436.7, [截止: 和 #60; 200]18) 建议她有 PA。她被转到横滨 Rosai 医院进一步评估高血压。体检是正常的身体质量指数 (23.4 公斤/米2)。血液检查正常, 包括 PAC (183 pg/毫升) 和 PRA (0.4 ng/毫升/小时)。盐水灌注试验显示微高 PAC (66 [截止: 和 #60; 60] pg/毫升)18, 表明她有轻度 PA。一个高 PAC 到 PRA 比率 ([146 pg/毫升]/[0.4 ng/毫升/小时] = 365 [截止: 和 #60; 200]) 后, 卡托普利的管理确认她有 PA (卡托普利挑战测试)18。CT 检测无明显肾上腺腺瘤 (图 5A)。为了确定醛固酮产生的肾上腺段 (s), 骑士与 ssAVS 进行了综合促肾上腺皮质激素刺激。在骑士队, PAC 或 PCC 在右和左中央静脉是 ([57600 pg/毫升]/[901 µg/dl] = 63.9) 和 ([1.8万 pg/毫升]/[389 µg/dl] = 46.3), 分别表明, 她有双边 PA (侧比率 = 1.4, 截止: 和 #60; 2.618, 表 1)。在右 ssAVS, PAC 在高级电视 (#1 在图 5B) 中, 中位电视 (#2)、侧电视 (#3) 和下位电视 (#4) 分别为 59100 pg/毫升、66400 pg/毫升、57300 pg/毫升、45400 pg/毫升。在左 ssAVS, PAC 在高级中值电视 (#1 在图 5C) 中, 上位电视 (#2)、侧电视 (#3) 和下位电视 (#4) 分别为 43900 pg/毫升、19600 pg/毫升、2.3万 pg/毫升、36900 pg/毫升。因此, PAC 超过 1.4万 pg/毫升, 在整个双边电视, 表明案件 #3 是真正的 IHA。她目前正接受皮质受体拮抗剂的治疗。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55716/55716fig1.jpg"> 
图 1:用于右骑士的导管.(A和B)导管的正面和侧面视图分别为 R 型。(C) 导管与 X 形状的正面视图。由图 1A和1C中的双向箭头表示的长度适合骑士的 "下腔静脉的长直径"15 。<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716fig1large.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/55716/55716fig2.jpg"> 
图 2:左骑士用导管.导管的正面观点与 L 形状。部分 #1、#2 和 #3 在图中分别适合下膈静脉和 LAV 的下腔、肾静脉和共同躯干, 让部分 #3 稳定地坐在共同的树干上。<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716fig2large.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/55716/55716fig3v4.jpg"> 
图 3:CT, 静脉造影, ssAVS, 组织学切除肾上腺 #1.(A) 对比度增强 CT。红色虚线的长度表示 "下腔静脉的长直径"15。红色和蓝色虚线之间的较大夹角表示 "雷夫的修正横向角"s = "xref" > 15。下腔静脉;Ao, 主动脉;孩子, 肾脏;T 肿瘤( B) 右肾上腺静脉造影。红色、黄色和粉红色箭头分别表示上级、侧面和劣质电视。( C、D、和E) 分别为侧向 (lat)、上级 (inf) 和劣质电视的静脉造影图像。粉红色、红色和黄色箭头指向的黑点表示微导管头。(F) 经切除的肾上腺皮质肿瘤 (T) 和相邻的肾上腺 (N) 的苏木素和曙红染色。(G和H)醛固酮合成酶 (CYP11B2: 缩写为 B2 在图中) 和类固醇11β羟化 (CYP11B1: B1) 的串行部分, 在图 3F。在 A 和 F H 中的刻度线分别表示1厘米和1毫米。<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55716/55716fig4.jpg"> 
图 4:CT, 静脉造影, ssAVS, 组织学切除肾上腺 #2.(A) 造影增强 CT. 下腔静脉;Ao, 主动脉;左肾上腺皮质肿瘤;右肾上腺皮质肿瘤(B) 左肾上腺静脉造影。黄色、红色和粉红色箭头分别显示中位、上侧和侧电视。静脉造影采用微导管, 其头部由一个黑色箭头表示。绿色箭头表示一个短的填补缺陷大概由于腺瘤。(C和D)分别进行外侧 (lat) 和侧向 (拉特) 电视的静脉造影图像。红色和粉红色箭头表示微导管头 (图 4C中的黑点和4D,分别)。值得注意的是, 在图 4B和数字 4C - 4D中, 同样的彩色箭头显示了相同的电视部分, 尽管中值电视的造影在图 4B中由黄色箭头表示, 但不可用。(F) 经切除的肾上腺皮质肿瘤 (T) 和相邻的肾上腺 (N) 的苏木素和曙红染色。(G和H)醛固酮合成酶 (CYP11B2: 缩写为 B2 在图中) 和类固醇11β羟化 (CYP11B1: B1) 的串行部分, 在图 4F。在 A 和 F H 中的刻度线分别表示1厘米和0.5 毫米。<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/55716/55716fig5.jpg"> 
图 5: CT 和静脉造影在 ssAVS 病例 #3.(A) 造影增强 CT. Rt, 它表示肾上腺。(B) 右肾上腺静脉造影。数字1、2、3和4与红色箭头表示上级, 中位, 横向和劣质电视. ( C) 左肾上腺静脉造影。数字1、2、3和4带有红色箭头, 表示中位、上侧、上侧和高级侧电视. 刻度线 = 1 厘米 (A)。<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Table 1" src="/files/ftp_upload/55716/55716table1v2.jpg"> 
表 1: 骑士和 ssAVS 案件数据 #1-3.<a href="http://ecsource.jove.com/files/ftp_upload/55716/55716table1v2large.jpg" target="_blank">请单击此处查看此表的较大版本.</a>

Watch this Scientific Journal Video about A Novel Method: Super-selective Adrenal Venous Sampling at JoVE.com

A Novel Method: Super-selective Adrenal Venous Sampling

' 超选择性 ' 肾上腺静脉取样 (ssAVS, 也称为节段性肾上腺静脉取样: sAVS) 是使用微导管来鉴别肾上腺段 (s) 产生过多的激素。本文对 ssAVS 技术进行了描述, 并对肾上腺节段性病变进行了 ssAVS 鉴定, 并探讨了 ssAVS 在今后肾上腺研究中的应用价值。

一种新的方法: 超选择性肾上腺静脉取样

Primary aldosteronism (PA) and subclinical Cushing&#39;s syndrome (SCS) are conditions in which the adrenal glands autonomously produce excessive amounts ...

a-novel-method-super-selective-adrenal-venous-sampling

Research

JoVE Journal

Medicine

23.2K 次观看.  Nerima Hikarigaoka Hospital. ' 超选择性 ' 肾上腺静脉取样 (ssAVS, 也称为节段性肾上腺静脉取样: sAVS) 是使用微导管来鉴别肾上腺段 (s) 产生过多的激素。本文对 ssAVS 技术进行了描述, 并对肾上腺节段性病变进行了 ssAVS 鉴定, 并探讨了 ssAVS 在今后肾上腺研究中的应用价值。

视频: A Novel Method: Super-selective Adrenal Venous Sampling

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

科研

医学

A Convenient Method for Extraction and Analysis with High-Pressure Liquid Chromatography of Catecholamine Neurotransmitters and Their Metabolites

The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and related diseases, but their precise measurement is still a challenge. Many protocols have been described for neurotransmitter measurement by a variety of instruments, including high-pressure liquid chromatography (HPLC). However, there are shortcomings, such as complicated operation or hard-to-detect multiple targets, which cannot be avoided, and presently, the dominant analysis technique is still HPLC coupled with sensitive electrochemical or fluorimetric detection, due to its high sensitivity and good selectivity. Here, a detailed protocol is described for the pretreatment and detection of catecholamines with high pressure liquid chromatography with electrochemical detection (HPLC-ECD) in real urine samples of infants, using electrospun composite nanofibers composed of polymeric crown ether with polystyrene as adsorbent, also known as the packed-fiber solid phase extraction (PFSPE) method. We show how urine samples can be easily precleaned by a nanofiber-packed solid phase column, and how the analytes in the sample can be rapidly enriched, desorbed, and detected on an ECD system. PFSPE greatly simplifies the pretreatment procedures for biological samples, allowing for decreased time, expense, and reduction of the loss of targets. 
Overall, this work illustrates a simple and convenient protocol for solid-phase extraction coupled to an HPLC-ECD system for simultaneous determination of three monoamine neurotransmitters (norepinephrine (NE), epinephrine (E), dopamine (DA)) and two of their metabolites (3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxy-phenylacetic acid (DOPAC)) in infants' urine. The established protocol was applied to assess the differences of urinary catecholamines and their metabolites between high-risk infants with perinatal brain damage and healthy controls. Comparative analysis revealed a significant difference in urinary MHPG between the two groups, indicating that the catecholamine metabolites may be an important candidate marker for early diagnosis of cases at risk for brain damage in infants.

We present a convenient solid-phase extraction coupled to high-pressure liquid chromatography (HPLC) with electrochemical detection (ECD) for simultaneous determination of three monoamine neurotransmitters and two of their metabolites in infants' urine. We also identify the metabolite MHPG as a potential biomarker for the early diagnosis of brain damage for infants.

用高压液相色谱法提取和分析儿茶酚胺神经递质及其代谢物的简便方法

The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and ...

化学

探索棕色脂肪组织中的代谢物交换

Arteriovenous Metabolomics to Measure In Vivo Metabolite Exchange in Brown Adipose Tissue

Brown adipose tissue (BAT) plays a crucial role in regulating metabolic homeostasis through a unique energy expenditure process known as non-shivering thermogenesis. To achieve this, BAT utilizes a diverse menu of circulating nutrients to support its high metabolic demand. Additionally, BAT secretes metabolite-derived bioactive factors that can serve as either metabolic fuels or signaling molecules, facilitating BAT-mediated intratissue and/or intertissue communication. This suggests that BAT actively participates in systemic metabolite exchange, an interesting feature that is beginning to be explored. Here, we introduce a protocol for in vivo mouse-level optimized BAT arteriovenous metabolomics. The protocol focuses on relevant methods for thermogenic stimulations and an arteriovenous blood sampling technique using Sulzer's vein, which selectively drains interscapular BAT-derived venous blood and systemic arterial blood. Next, a gas chromatography-based metabolomics protocol using those blood samples is demonstrated. The use of this technique should expand the understanding of BAT-regulated metabolite exchange at the inter-organ level by measuring the net uptake and release of metabolites by BAT.

作者聚焦:阐明脂肪组织代谢和疾病预防的新途径 

In this protocol, methods relevant for BAT-optimized arteriovenous metabolomics using GC-MS in a mouse model are outlined. These methods allow for the acquisition of valuable insights into BAT-mediated metabolite exchange at the organismal level.

动静脉代谢组学测量棕色脂肪组织 中的体内 代谢物交换

Brown adipose tissue (BAT) plays a crucial role in regulating metabolic homeostasis through a unique energy expenditure process known as non-shivering ...

生物化学

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful compared to other more aggressive methods such as retro-orbital plexus sample collection. However, this blood collection method is sometimes limited by an unsatisfactory success rate. Here, we introduce a method for blood collection through the subclavian vein puncture. The subclavian vein is located just under the clavicle and this vein is large enough to fulfill the volume requirements of blood collection. Our results show that this method is safe and applicable for blood collection sampling with the required blood volume. Blood collection through the subclavian vein puncture could serve as an alternative blood collection method in case of failed tail vein blood sampling in rats.

Here, we present a protocol to collect blood samples from the rat subclavian vein.

锁骨下静脉穿刺作为大鼠采血的一种替代方法

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...

Confocal Microscopy to Measure Three Modes of Fusion Pore Dynamics in Adrenal Chromaffin Cells

Dynamic fusion pore opening and closure mediate exocytosis and endocytosis and determine their kinetics. Here, it is demonstrated in detail how confocal microscopy was used in combination with patch-clamp recording to detect three fusion modes in primary culture bovine adrenal chromaffin cells. The three fusion modes include 1) close-fusion (also called kiss-and-run), involving fusion pore opening and closure, 2) stay-fusion, involving fusion pore opening and maintaining the opened pore, and 3) shrink-fusion, involving shrinkage of the fusion-generated &#937;-shape profile until it merges completely at the plasma membrane.
To detect these fusion modes, the plasma membrane was labeled by overexpressing mNeonGreen attached with the PH domain of phospholipase C &#948; (PH-mNG), which binds to phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) at the cytosol-facing leaflet of the plasma membrane; vesicles were loaded with the fluorescent false neurotransmitter FFN511 to detect vesicular content release; and Atto 655 was included in the bath solution to detect fusion pore closure. These three fluorescent probes were imaged simultaneously at ~20-90 ms per frame in live chromaffin cells to detect fusion pore opening, content release, fusion pore closure, and fusing vesicle size changes. The analysis method is described to distinguish three fusion modes from these fluorescence measurements. The method described here can, in principle, apply to many secretory cells beyond chromaffin cells.

This protocol describes a confocal imaging technique to detect three fusion modes in bovine adrenal chromaffin cells. These fusion modes include 1) close-fusion (also called kiss-and-run), involving fusion pore opening and closure, 2) stay-fusion, involving fusion pore opening and maintaining the opened pore, and 3) shrink-fusion, involving fused vesicle shrinkage.

共聚焦显微镜测量肾上腺嗜铬细胞融合孔隙动力学的三种模式

Dynamic fusion pore opening and closure mediate exocytosis and endocytosis and determine their kinetics. Here, it is demonstrated in detail how confocal ...

神经科学

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. The method offers a non-invasive option to assess long term patterns in both domestic and free ranging horses. This protocol describes the enzyme linked immunoassay involved and the associated biochemical validation.

糖皮质激素粪分析:在马属动物无创监测肾上腺

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

生物学

同时记录脑电图的大鼠脑脊液和血液

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats During EEG Recordings

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

作者聚焦:获取高质量的 CSF 和血液样本以发现癫痫生物标志物 

The protocol shows repeated cerebrospinal fluid and blood collections from epileptic rats performed in parallel with continuous video-electroencephalogram (EEG) monitoring. These are instrumental for exploring possible links between changes in various body fluid molecules and seizure activity.

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many ...

一种新的方法: 超选择性肾上腺静脉取样

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