作者没有什么可透露的。

本研究获得了上海九人民医院伦理委员会的伦理认同和每位患者的书面知情同意。
1. 病人和外科材料
备注:根据欧洲泌尿外科协会的指导原则, 确诊为 BPH 并需要手术治疗的患者可包括10。这种手术过程需要经验和长时间的学习曲线。外科医生的经验是完成这项技术的最重要的因素之一, 特别是前列腺的体积超过100克。钬激光微爆炸效应适用于摘除术。
<ol><li>手术前准备<ol>
<li>根据《香水指南》10, 评估需要手术治疗的前列腺增生患者。</li>
		<li>排除那些具有以下条件: 严重泌尿系感染, 严重肺部或心脏病, 肾功能损害, 严重出血紊乱, 血压控制不佳, 神经源性膀胱功能障碍, 膀胱癌, 前前列腺手术, 前列腺癌, 尿道狭窄, 和其他手术禁忌在我们以前的报告中提到的11。</li>
		<li>根据结果进行术前评估并进行一定的围手术期治疗。</li>
		<li>对病人施行腰椎或全身麻醉, 并将病人置于碎石位置。</li>
	</ol>
</li>
	<li>手术前评估 备注:对于术前评估, 执行以下分析包括常规手术检查: 尿流动力学分析, 前列腺超声分析, 前列腺特异抗原 (PSA) 水平分析和 urethroscopy 分析。<ol>
<li>使用550µm 端发射激光光纤和100瓦特脉冲钬激光器执行计画, 电源设置为80-100 笏 1.6-2 J/秒和 40-50 Hz. 用机械 morcellator 执行经尿道 morcellation 与 26 F 肾。用1升的生理盐水 (0.9%) 作为灌溉ﬂuid。</li>
	</ol>
</li>
</ol>2. 摘除程序
备注:使用等离子灭菌消毒仪器, 并将仪器放入无菌包装中, 直至使用。对患者进行腰椎或全身麻醉, 并在手术期间将病人置于碎石位置。
<ol><li>前列腺和尿道的观察<ol>
<li>插入连续流膀胱, 确保组织损伤最小, 并仔细观察前列腺、尿道和膀胱。</li>
		<li>用视觉闭孔定位输尿管口以进行一般评估, 包括中位叶容积和膀胱组织数量。</li>
		<li>确定 verumontanum 是摘除术中最重要的地标之一。将牙科与激光机连接, 激光机的功率设置为 80-100 W, 每秒 1.6-2 J 和40-50 赫兹。</li>
	</ol>
</li>
	<li>隧道切口周围的6点的位置前列腺与患者在碎石位置。<ol>
<li>使用钬激光, 在 verumontanum 前面的6点位置进行短水平切口 (如图 1A所示), 以定位前列腺手术胶囊。</li>
		<li>将前列腺手术胶囊从 verumontanum 暴露到膀胱颈部。在手术胶囊的层上, 创建一个隧道, 从顶端向膀胱颈部后部, 如图 1A和1D所示。</li>
		<li>当手术接近膀胱颈部时, 确保粘膜和肌肉得到保护, 以促进有效的尿液控制。</li>
	</ol>
</li>
	<li>侧瓣移除 备注:该胶囊的特点是光滑的白色组织与足够的血管在表面上。出血血管也可以用比无核过程更远的距离 (更大的距离适用于止血, 而较小的距离适合切割) 的激光来封闭。在手术过程中, 应毫不犹豫地回到前列腺先端, 以确保确切的手术胶囊位于, 这有助于避免在手术中迷路, 当操作员找不到清晰的视野。<ol>
<li>首先, enucleate 左外侧叶从6点到12点位置在手术胶囊的水平, 然后 enucleate 其他侧瓣类似。</li>
		<li>在该过程中, 使用范围鞘结合激光切割的模糊分离。</li>
		<li>将扩大的前列腺挂在12点的膀胱颈部位置, 而不减少。</li>
		<li>从膀胱颈部切开组织, 在摘除术结束时将其推入膀胱。</li>
		<li>检查前列腺胶囊, 确保不发生活动出血。移除所有放大的腺体并停止严重出血点 (图 1A-1F中所示)。</li>
	</ol>
</li>
</ol>3. Morcellation
备注:有二个齿轮在 morcellator, 可以根据脚压力被转移应用。第一齿轮采用轻压吸收组织和水, 而第二齿轮用于 morcellation。为了预防并发症, 提高程序的效率, 我们开发了传统的 morcellation 技能和程序。在本例中, 我们在前面的研究12中详细讨论了实现的方法。从膀胱取出的组织显示在图 2中。
<ol><li>Morcellation 程序<ol>
<li>根据 morcellator 指令装备 morcellator 和火柴管, 无论使用何种 morcellator。有效使用 morcellator 从膀胱中取出前列腺组织。</li>
		<li>在手术过程中, 要非常小心地治疗膀胱组织13。在膀胱内保持足够的灌溉水, 以保持膀胱肿胀, 避免膀胱损伤, 并保持 morcellator 远离膀胱壁, 以最大限度地保护膀胱壁。</li>
		<li>最后, 观察膀胱的一般情况, 确保膀胱内不留有保留的组织。</li>
	</ol>
</li>
	<li>Morcellation 技能<ol>
<li>正如我们早期研究报告的12, 在 morcellation 之前, 评估腺体组织的质地。</li>
		<li>用激光来挖孔, 以提供与腺体表面的摩擦, 从而获得平滑的表面。</li>
		<li>使用逆技术 morcellate 大的和牢固的组织以及纤维化, 球形腺体与光滑的表面和牢固的组织。在过程中逆转 morcellator 为它的开头面对向下在膀胱。确保循环水畅通, 在 morcellation 期间不断感受膀胱压力。</li>
	</ol>
</li>
</ol>4. 术后治疗
<ol style="margin-left: 40px;"><li>手术后, 插入导管, 然后将生理盐水冲洗到膀胱内, 直到灌溉水清澈或微红。</li>
	<li>取出导管, 直到尿液变得清晰而不出现严重血尿。</li>
	<li>如果观察不到明显的保留, 病人可以在导尿管取出后排尿, 则从医院释放病人14,15。</li>
</ol>5. 后续观察
<ol><li>出院后, 检查患者的随访数据 1, 6, 12 月, 其中包括国际前列腺症状评分 (IPSS), 过度活跃膀胱评分系统 (OABSS), 生活质量 (质量) 和 Qmax。如果发生术后并发症, 请应用进一步治疗14。</li>
</ol>

<ol>
	<li>Bushman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiology,+epidemiology,+and+natural+history+of+benign+prostatic+hyperplasia.">Etiology, epidemiology, and natural history of benign prostatic hyperplasia.</a> Urol Clin North Am. 36 (4), 403-415 (2009).</li><li>Millin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retropubic+prostatectomy;+a+new+extravesical+technique;+report+of+20+cases.">Retropubic prostatectomy; a new extravesical technique; report of 20 cases.</a> Lancet. 2 (6380), 693-696 (1945).</li><li>Kahokehr, A. A., Gilling, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Which+laser+works+best+for+benign+prostatic+hyperplasia?.">Which laser works best for benign prostatic hyperplasia?.</a> Curr Urol Rep. 14 (6), 614-619 (2013).</li><li>Gilling, P. J., Cass, C. B., Malcolm, A. R., Fraundorfer, 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=Combination+holmium+and+Nd:YAG+laser+ablation+of+the+prostate:+initial+clinical+experience.">Combination holmium and Nd:YAG laser ablation of the prostate: initial clinical experience.</a> J Endourol. 9 (2), 151-153 (1995).</li><li>Fraundorfer, M. R., Gilling, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Holmium:YAG+laser+enucleation+of+the+prostate+combined+with+mechanical+morcellation:+preliminary+results.">Holmium:YAG laser enucleation of the prostate combined with mechanical morcellation: preliminary results.</a> Eur Urol. 33 (1), 69-72 (1998).</li><li>Chen, Y. B., 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=A+prospective,+randomized+clinical+trial+comparing+plasmakinetic+resection+of+the+prostate+with+holmium+laser+enucleation+of+the+prostate+based+on+a+2-year+followup.">A prospective, randomized clinical trial comparing plasmakinetic resection of the prostate with holmium laser enucleation of the prostate based on a 2-year followup.</a> J Urol. 189 (1), 217-222 (2013).</li><li>Kahokehr, A., Gilling, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enucleation+techniques+for+benign+prostate+obstruction:+which+one+and+why?.">Enucleation techniques for benign prostate obstruction: which one and why?.</a> Curr Opin Urol. 24 (1), 49-55 (2014).</li><li>Kuntz, R. M., Lehrich, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transurethral+holmium+laser+enucleation+versus+transvesical+open+enucleation+for+prostate+adenoma+greater+than+100+gm.::+a+randomized+prospective+trial+of+120+patients.">Transurethral holmium laser enucleation versus transvesical open enucleation for prostate adenoma greater than 100 gm.:: a randomized prospective trial of 120 patients.</a> J Urol. 168 (4 Pt 1), 1465-1469 (2002).</li><li>Elzayat, E., Habib, E., Elhilali, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Holmium+laser+enucleation+of+the+prostate+in+patients+on+anticoagulant+therapy+or+with+bleeding+disorders.">Holmium laser enucleation of the prostate in patients on anticoagulant therapy or with bleeding disorders.</a> J Urol. 175 (4), 1428-1432 (2006).</li><li>Oelke, 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=EAU+guidelines+on+the+treatment+and+follow-up+of+non-neurogenic+male+lower+urinary+tract+symptoms+including+benign+prostatic+obstruction.">EAU guidelines on the treatment and follow-up of non-neurogenic male lower urinary tract symptoms including benign prostatic obstruction.</a> Eur Urol. 64 (1), 118-140 (2013).</li><li>Xu, 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=Efficacy+and+safety+of+an+improved+technique+of+holmium+laser+enucleation+of+the+prostate+versus+open+prostatectomy+in+treating+enlarged+prostates+combined+with+multiple+bladder+stones:+one-year+of+followup.">Efficacy and safety of an improved technique of holmium laser enucleation of the prostate versus open prostatectomy in treating enlarged prostates combined with multiple bladder stones: one-year of followup.</a> Int J Clin Exp Med. 9 (12), 23612-23617 (2016).</li><li>Chen, Q., 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=An+improved+morcellation+procedure+for+holmium+laser+enucleation+of+the+prostate.">An improved morcellation procedure for holmium laser enucleation of the prostate.</a> J Endourol. 26 (12), 1625-1628 (2012).</li><li>El Tayeb, M. M., Borofsky, M. S., Paonessa, J. E., Lingeman, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wolf+Piranha+Versus+Lumenis+Versacut+Prostate+Morcellation+Devices:+A+Prospective+Randomized+Trial.">Wolf Piranha Versus Lumenis Versacut Prostate Morcellation Devices: A Prospective Randomized Trial.</a> J Urol. , (2015).</li><li>Gilling, P. J., Aho, T. F., Frampton, C. M., King, C. J., Fraundorfer, 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=Holmium+laser+enucleation+of+the+prostate:+results+at+6+years.">Holmium laser enucleation of the prostate: results at 6 years.</a> Eur Urol. 53 (4), 744-749 (2008).</li><li>Shah, H. N., 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=Prospective+evaluation+of+the+learning+curve+for+holmium+laser+enucleation+of+the+prostate.">Prospective evaluation of the learning curve for holmium laser enucleation of the prostate.</a> J Urol. 177 (4), 1468-1474 (2007).</li><li>Gratzke, 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=EAU+Guidelines+on+the+Assessment+of+Non-neurogenic+Male+Lower+Urinary+Tract+Symptoms+including+Benign+Prostatic+Obstruction.">EAU Guidelines on the Assessment of Non-neurogenic Male Lower Urinary Tract Symptoms including Benign Prostatic Obstruction.</a> Eur Urol. 67 (6), 1099-1109 (2015).</li><li>Vincent, M. W., Gilling, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HoLEP+has+come+of+age.">HoLEP has come of age.</a> World J Urol. 33 (4), 487-493 (2015).</li><li>Qian, X., 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=Functional+outcomes+and+complications+following+B-TURP+versus+HoLEP+for+the+treatment+of+benign+prostatic+hyperplasia:+a+review+of+the+literature+and+Meta-analysis.">Functional outcomes and complications following B-TURP versus HoLEP for the treatment of benign prostatic hyperplasia: a review of the literature and Meta-analysis.</a> Aging Male. 20 (3), 184-191 (2017).</li><li>Xu, H., Chen, Y. B., Gu, M., Chen, Q., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+noise+hazard+during+the+holmium+laser+enucleation+of+prostate.">Evaluation of noise hazard during the holmium laser enucleation of prostate.</a> BMC Urol. 17 (1), 71 (2017).</li><li>Gilling, P. J., 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=Long-term+results+of+a+randomized+trial+comparing+holmium+laser+enucleation+of+the+prostate+and+transurethral+resection+of+the+prostate:+results+at+7+years.">Long-term results of a randomized trial comparing holmium laser enucleation of the prostate and transurethral resection of the prostate: results at 7 years.</a> BJU Int. 109 (3), 408-411 (2012).</li></ol>

本研究得到上海市浦东卫生局重点学科群建设项目 (PWZxq2014-11)、优秀医学学术带头人方案、中西医结合专项建设的支持。Wedicine 上海综合医院 (ZHYY-ZXYJHZX-1-03)。

在本议定书中, 我们介绍了前列腺增生症的外科治疗方法。计画可以在日常的临床实践中使用。如前所述, 前列腺增生手术的发展包括三个地标。第一个是开放性手术。根治前列腺切除术和耻骨上开放性前列腺切除术在临床上有广泛的应用。尽管如此, 这种技术还是有严重的不良副作用, 在围手术期经常会出现。这些副作用包括出血, 手术后的大疤痕, 缓慢的病人恢复。此外, 手术领域仅限于骨盆腔?...

良性前列腺增生 (BPH) 是前列腺增生, 发生在老年男性。这种情况开始在大约 40 y 老, 并且发病率可能到达 50%-60% 为男性在他们的60s 和 80%-90% 为男性在他们的70s 和 80s1。BPH 导致渐进性梗阻, 下尿路症状, 保留及其并发症, 甚至肾衰竭, 所有这些都降低了老年男性的生活质量。经医学治疗后, 外科治疗常被用作症状性前列腺增生症的最有效治疗方法。
为治疗前列腺增生症的症状, 包括在 1894年2和经尿道内窥镜手术 (前列腺切除术) 的第一次开放性前列腺摘除术, 已开发了各种类型的手术。经尿道前列腺切除术自其发展以来, 已被认为是手术纾缓扩张的金标准。
钬激光的波长为 2120 nm, 类似于组织水的3, 它会诱发水的汽化。组织穿透率达0.4 毫米, 凝血也可与之一起操作。该过程最初是在 1994年4中为 BPH 引入的。在 1998年, 彼得针梳机使用该设备执行摘除应用程序5。正如彼得针梳机等人在二十世纪九十年代4中提出的那样, 钬激光摘除前列腺 (计画) 是用来治疗前列腺增生症的两个早期类型的手术, 包括开放摘除和经尿道前列腺切除术4。随着增生过程的发展, 前列腺周围区由过渡区围困, 形成所谓的手术胶囊。计画允许切除前列腺的增生部分, 分离前列腺胶囊。因此, 计画, 作为解剖手术, 可以 enucleate 扩大的前列腺区从形成分离之间的间隔和胶囊。与传统的手术方法相比, 计画可能更安全、更有效、更彻底, 并具有持久的效果。与经尿道前列腺切除术相比, 钬降低了手术时间、灌水时间、导尿管的时机, 缩短了住院天数。6,7,8该技术还可以安全地执行抗凝治疗或出血紊乱的患者9。虽然计画有不同的程序, 但主要的程序与下面提到的是相同的。我们还介绍了我们自己的经验和这种方法的创新, 以使它更容易接受和缩短学习曲线。该程序特别适用于大前列腺和那些结合膀胱结石, 鉴于钬激光也可以用于碎石术8。本文介绍了该技术及具体程序。

<table border="0" cellpadding="0" cellspacing="0" width="1069">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:260px;">holmium laser</td>
			<td style="width:205px;">Lumenis, USA</td>
			<td style="width:344px;"></td>
			<td style="width:260px;">PowerSuite 100W</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">mechanical morcellator</td>
			<td>Lumenis, USA</td>
			<td></td>
			<td>VersaCut</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">27Fr continuous flow resectoscope</td>
			<td>Storz, Tuttlingen, Germany</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

surgical treatment for benign prostatic hyperplasia: holmium laser enucleation of the prostate (holep).

良性前列腺增生 (BPH) 通常发生在老年男性。病情导致下尿路症状, 从而降低生活质量。前列腺增生症包括2主要治疗: 药物和手术。外科治疗通常是最有效和最后的干预。钬激光摘除前列腺 (计画), 其中一个最有效的手术程序, BPH, 是进行 transurethrally。在外科手术中, 最重要和最困难的部分是定位前列腺手术胶囊, 这与解剖手术的理念是一致的。治疗前列腺不同部位可能需要不同的技能。计画具有良好的止血性能, 能够治疗膀胱结石, 这可能是前列腺增生症的并发症。该技术特别适用于不同体积和大小的前列腺。事实上, 计画也提出了一些缺点, 如长时间学习曲线和昂贵的设备。无论如何, 这种方法可能是 "新标准" 和优秀的手术治疗前列腺增生症的方法。

回顾性数据, 收集从2015年到 2016, 是从我的临床数据库的部门。本研究共招收100例前列腺增生症患者。围手术期结果包括: 术后时间 (OT)、术后血红蛋白减少、Na+变异、住院持续时间、膀胱冲洗时间和随访数据。在我们的研究中也收集了基本的病人特征。此外, 还注意到插管时间和住院时间。术后1、4、24周采集随访资料和术后尿并发症。所有数据都显示在表 1</...

Watch this Scientific Journal Video about Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate HoLEP. at JoVE.com

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate HoLEP.

在这里, 我们提出一个安全和有效的协议, 钬激光摘除前列腺, 治疗良性前列腺增生。

良性前列腺增生的外科治疗: 钬激光前列腺摘除术 (计画)。

Benign prostatic hyperplasia (BPH) commonly occurs among elderly males. The condition causes symptoms in the lower urinary tract, thereby reducing the ...

surgical-treatment-for-benign-prostatic-hyperplasia-holmium-laser

Research

JoVE Journal

Medicine

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate (HoLEP).

16.8K 次观看.  Shanghai Jiaotong University School of Medicine. 在这里, 我们提出一个安全和有效的协议, 钬激光摘除前列腺, 治疗良性前列腺增生。

视频: Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate HoLEP.

Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma 1,2. The dissected cells can be used for a variety of investigations, such as transcriptomic or proteomic studies, DNA assessment or chromosomal analysis 2,3. An especially challenging application of LMD is transcriptome analysis, which, due to the lability of RNA 4, can be particularly prominent when cells are dissected from tissues that are rich of RNases, such as the pancreas. A microdissection protocol that enables fast identification and collection of target cells is essential in this setting in order to shorten the tissue handling time and, consequently, to ensure RNA preservation. 
Here we describe a protocol for acquiring human pancreatic beta cells from surgical specimens to be used for transcriptomic studies 5. Small pieces of pancreas of about 0.5-1 cm3 were cut from the healthy appearing margins of resected pancreas specimens, embedded in Tissue-Tek O.C.T. Compound, immediately frozen in chilled 2-Methylbutane, and stored at -80 &deg;C until sectioning. Forty serial sections of 10 &mu;m thickness were cut on a cryostat under a -20 &deg;C setting, transferred individually to glass slides, dried inside the cryostat for 1-2 min, and stored at -80 &deg;C. 
Immediately before the laser microdissection procedure, sections were fixed in ice cold, freshly prepared 70% ethanol for 30 sec, washed by 5-6 dips in ice cold DEPC-treated water, and dehydrated by two one-minute incubations in ice cold 100% ethanol followed by xylene (which is used for tissue dehydration) for 4 min; tissue sections were then air-dried afterwards for 3-5 min. Importantly, all steps, except the incubation in xylene, were performed using ice-cold reagents - a modification over a previously described protocol 6. utilization of ice cold reagents resulted in a pronounced increase of the intrinsic autofluorescence of beta cells, and facilitated their recognition. For microdissection, four sections were dehydrated each time: two were placed into a foil-wrapped 50 ml tube, to protect the tissue from moisture and bleaching; the remaining two were immediately microdissected. This procedure was performed using a PALM MicroBeam instrument (Zeiss) employing the Auto Laser Pressure Catapulting (AutoLPC) mode. The completion of beta cell/islet dissection from four cryosections required no longer than 40-60 min. Cells were collected into one AdhesiveCap and lysed with 10 &mu;l lysis buffer. Each single RNA specimen for transcriptomic analysis was obtained by combining 10 cell microdissected samples, followed by RNA extraction using the Pico Pure RNA Isolation Kit (Arcturus). This protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their rapid and accurate recognition and collection. Further improvement of this procedure could enable the dissection of phenotypically different beta cells, with possible implications for better understanding the changes associated with type 2 diabetes.

Laser microdissection is a technique that allows the recovery of selected cells from minute amounts of parenchyma. Here we describe a protocol for acquiring human pancreatic islets from surgical specimens to be used for transcriptomic studies. Our protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their collection.

从手术标本的人类胰岛细胞激光显微切割的改进方法

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma 1,2. The dissected ...

科研

医学

Renal Capsule Xenografting and Subcutaneous Pellet Implantation for the Evaluation of Prostate Carcinogenesis and Benign Prostatic Hyperplasia

New therapies for two common prostate diseases, prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend critically on experiments evaluating their hormonal regulation. Sex steroid hormones (notably androgens and estrogens) are important in PrCa and BPH; we probe their respective roles in inducing prostate growth and carcinogenesis in mice with experiments using compressed hormone pellets. Hormone and/or drug pellets are easily manufactured with a pellet press, and surgically implanted into the subcutaneous tissue of the male mouse host. We also describe a protocol for the evaluation of hormonal carcinogenesis by combining subcutaneous hormone pellet implantation with xenografting of prostate cell recombinants under the renal capsule of immunocompromised mice. Moreover, subcutaneous hormone pellet implantation, in combination with renal capsule xenografting of BPH tissue, is useful to better understand hormonal regulation of benign prostate growth, and to test new therapies targeting sex steroid hormone pathways.

We describe the manufacture of compressed hormone pellets, as well as subcutaneous surgical implantation into mice. This strategy can be combined with the growth of cell and tissue xenografts under the renal capsule of athymic nude mice to evaluate hormonal carcinogenesis and regulation of benign prostate growth.

肾小囊和异种移植皮下颗粒植入前列腺癌发生和良性前列腺增生症的评价

New therapies for two common prostate diseases,  prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend  critically on experiments ...

Laser-capture Microdissection of Human Prostatic Epithelium for RNA Analysis

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types. Healthy prostate is comprised of fibromuscular stroma surrounding discrete epithelial-lined secretory lumens and a very small population of immune and neuroendocrine cells. In contrast, areas of prostate cancer have increased dysplastic luminal epithelium with greatly reduced or absent stromal population. Given the profound difference between stromal and epithelial cell types, it is imperative to separate the cell types for any type of downstream molecular analysis. Despite this knowledge, the bulk of gene expression studies compare benign prostate to cancer without micro-dissection, leading to stromal bias in the benign samples. Laser-capture micro-dissection (LCM) is an effective method to physically separate different cell types from a specimen section. The goal of this protocol is to show that RNA can be successfully isolated from LCM-collected human prostatic epithelium and used for downstream gene expression studies such as RT-qPCR and RNAseq.

The goal of this protocol is to use laser-capture micro-dissection as an effective method to isolate pure populations of cell types from heterogeneous prostate tissues for downstream RNA analysis.

人类前列腺上皮的激光捕获显微切割的RNA分析

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types.  Healthy prostate is comprised of ...

Murine Prostate Micro-dissection and Surgical Castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

鼠前列腺微解剖和手术去势

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

在耐药性颞叶癫痫患者卵圆孔电极记录的网络分析

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Transcanalicular Diode Laser-assisted Dacryocystorhinostomy for the Treatment of Primary Acquired Nasolacrimal Duct Obstruction

Today&#39;s gold standard in the treatment of infrasaccal primary acquired nasolacrimal duct obstruction (PANDO) is external dacryocystorhinostomy (DCR), a relatively invasive procedure that can be performed after failure of recanalizing treatments. However, with progress in the field of diode laser technology, new approaches have emerged. Laser-assisted transcanalicular DCR with subsequent bicanalicular silicon intubation is a new option showing great promise as a viable minimally invasive procedure. Under permanent endoscopic visual control from the nasal cavity, a diode laser fiber is inserted into the lacrimal sac and laser energy is applied to create a bony ostium between the lacrimal sac and the nasal cavity. Since no skin incision needs to be made, advantages of this method comprise the sparing of the skin as well as the medial palpebral structures and the physiological palpebral-canalicular pump mechanism. The duration of surgery as well as reconvalescence is generally shorter than with external DCR. Complications include silicon tube prolapse, mild swelling and, rarely, canalicular infection and thermal injury. One-year functional success rates, defined as complete resolution of symptoms and ostium patency, are high, yet still range behind those of external DCR. However, secondary external DCR after failure of laser-assisted DCR can be performed without difficulty. Thus, laser-assisted transcanalicular DCR is a valid option that should be considered as a second-step procedure after failure of recanalization procedures and before external DCR.

The goal of this protocol is to present transcanalicular laser-assisted dacryocystorhinostomy as a minimally invasive approach in the treatment of primary acquired nasolacrimal duct obstruction.

Transcanalicular 二极管激光辅助泪囊鼻腔吻合术治疗原发性鼻泪管阻塞

Today&#39;s gold standard in the treatment of infrasaccal primary acquired nasolacrimal duct obstruction (PANDO) is external dacryocystorhinostomy (DCR), ...

Photoselective Vaporesection of the Prostate via an End-firing Lithium Triborate Crystal Laser

The occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem with a high incidence in the aging male population. Although it is not a life-threatening disease, BPH causes problems that seriously impact the quality of life. Here, we introduce a new technique called photoselective vaporesection of the prostate (PVRP) in treating BPH, which can be seen as a variation of photoselective vaporization of the prostate (PVP). This procedure presents several advantages compared to the PVP technique including less laser energy loss, less intraoperative complications as well as more tissue resection rate.

Here we present a protocol to perform Photoselective Vaporesection of the Prostate (PVRP) for benign prostatic hyperplasia (BPH) treatment.

选择性绿激光锂三硼酸晶体激光 Vaporesection 前列腺的研究

The occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem with a high incidence in the aging ...

Enucleation of the Prostate for the Treatment of Benign Prostatic Hyperplasia Using a 980 nm Diode Laser

In the aging male population, the occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem. Here, we introduce a new technique called 980 nm diode laser enucleation (DiLEP) to treat BPH1. Diode lasers can absorb both water and hemoglobin at the same time, so they are good for cutting and hemostasis2. The diode laser was approved by the FDA in 2007, and has been used in the treatment of BPH because of its effective cutting and hemostasis effect3. DiLEP presents several advantages over other techniques, such as TURP, HoLEP, and PVP. During the procedure, we define the boundary of a high-volume prostate and separate it into three lobes with a diode laser by burning two rings and one groove (like a Cupid&#39;s arrow). Compared to other procedures, mDiLEP has fewer intraoperative complications, a shorter learning curve, and achieves more tissue resection.

Here, we present a protocol for modified 980 nm diode laser enucleation to treat large volume benign prostatic hyperplasia.

使用 980 nm 二极管激光进行前列腺摘除术治疗良性前列腺增生

In the aging male population, the occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem. ...

Diagnosis and Surgical Treatment of Human Brucellar Spondylodiscitis

Brucellar spondylodiscitis (BS) is the most prevalent and significant osteoarticular presentation of human Brucellosis, which is commonly manifested in pastoral communities. It is difficult to differentially diagnose and usually leads to irreversible neurologic deficits and spinal deformities. The initial diagnosis of BS is based on clinical findings and radiographic assessments, and the confirmed diagnosis should be established by the isolation of Brucella species from the blood and/or the standard tube agglutination test. Differential diagnosis of multifocal BS from either degenerative disc diseases or tuberculosis is especially highlighted. The surgical approach, either endoscopic or open, is demonstrated in detail, accompanied by radiographic evidence of structural compression or severe instability. Further, the crucial surgical steps, including single-stage transforaminal decompression, debridement, interbody fusion, and internal fixation, are explained. Moreover, perioperative care and postoperative rehabilitation are also addressed. Taken together, this clinical algorithm presents a practical guide that has yielded substantially satisfactory outcomes in the past decades, which can also be introduced for large-scale application to manage human BS, especially in endemic regions.

We describe a clinical algorithm, based on decades of front-line experience of diagnosis and surgical treatment of human Brucellar spondylodiscitis in the largest medical center of the China's Xinjiang Pastoral Area.

人类布鲁塞拉性皮诺皮诺皮炎的诊断和手术治疗

Brucellar spondylodiscitis (BS) is the most prevalent and significant osteoarticular presentation of human Brucellosis, which is commonly manifested in ...

Focal Laser Ablation of Prostate Cancer: An Office Procedure

In this article, we describe and illustrate an outpatient procedure for focal laser ablation (FLA) of prostate cancer (PCa). The procedure is conceptually similar to a fusion biopsy and is performed under local anesthesia in a clinic setting; treatment time is usually less than one hour. Laser insertion is guided by ultrasound; lesion targeting is via magnetic resonance imaging-ultrasound (MRI/US) fusion, as in targeted prostate biopsy. Real-time ablation monitoring is achieved utilizing a thermal probe adjacent to the laser fiber. The video demonstrates procedure planning, patient preparation, various steps during the procedure, and treatment monitoring. Safety, feasibility, and efficacy of this approach have been established during a previous trial. Outpatient FLA under local anesthesia is an option for management of intermediate risk prostate cancer.

This article presents and describes an outpatient treatment for prostate cancer using focal laser ablation. Laser catheter placement is guided by MRI-ultrasound fusion imaging in a fashion similar to prostate needle biopsy. Treatment is monitored in real-time with a thermal probe, placed adjacent to the laser fiber.

前列腺癌聚焦激光消融:办公室程序

In this article, we describe and illustrate an outpatient procedure for focal laser ablation (FLA) of prostate cancer (PCa). The procedure is conceptually ...