没有资金来源。

1。在胎盘娩出的制备<ol><li>标签（第一出生）或两个（第二个出生的）夹具的双胞胎的脐带。 </li><li>检查胎盘的完整性或中断的孕产妇和胎儿的表面。 </li><li>记录以下数据：型线插入（中央，偏心，边际或velamentous），在脐带（通常是一个静脉和动脉，有时只有一条动脉）和两个胎盘股之间的颜色差异的血管的数目。除以膜的一节可以送病理检查，以确认chorionicity类型。 </li><li>胎盘就可以被放置在一个塑料碗和冷藏，直到期末考试（最好的一个星期内）和彩色染料注射。 </li><li>胎盘不得冻结或固定（不使用福尔马林）。 </li></ol> 2。导尿脐船只<ol><li>用温水或生理盐水洗净胎盘。 </li><li>修剪周围的膜，除去除以跨双膜关闭amnions和剥离（更好的血管吻合和更优质的胎盘图片的可视化）。 </li><li>每个脐带断面线插入约5厘米的距离。 </li><li>轻轻挤压出来，从脐带船只和胎盘血管的血凝块。 </li><li>通常很容易识别由于其较大的直径相比，直径较小的两个脐动脉脐静脉。 </li><li> Cannulate一个适当大小的导管脐静脉。避免虚假的段落。 </li><li> Cannulate一个较小的导管脐动脉。使用镊子，以扩大脐动脉管腔。避免虚假的段落。 2脐动脉只有一个需要插管以来吻合（Hyrtl）连接线插入附近2动脉。 </li><li>其他脐带重复这两个步骤。 </li><li>导管的位置，可以促进轻轻的来回按摩脐船只。此过程可用于任何类型的导管。我们选择使用（回收）我们的新生儿病房用于在新生儿脐带导尿的导管。 </li><li>扎两个线左右的磁带片，以避免流动的彩色染料在染料注射。 </li></ol> 3。注射用有色染料<ol><li>连接20 ml注射器充满彩色染料每个导管。 </li><li>可用于任何粘性的彩色染料，以可视化的胎盘血管架构。使用对比色，让良好的可视化的吻合（深颜色的动脉，静脉鲜艳的色彩）。 </li><li>轻轻注入（低压）的彩色染料，同时在静脉助理轻轻推染料让有色染料来填补所有的胎盘血管，也是最小的。 </li><li>要特别注意血管赤道附近的小血管（血管赤道两张吻合的地方相互连接）。 </li><li>重复前面的步骤注入动脉彩色染料。地注意到：动脉注入和需要更多的耐心可能会更加困难。 </li><li>其他脐带重复这两个步骤。 </li></ol> 4。彩色染料注射后的评估和胎盘的文件<ol><li>仔细检查的血管赤道和记录吻合的数量和类型。 </li><li>胎盘上放置一个卷尺来测量的数码照片上的直径和胎盘股份。 </li><li>使用一个高分辨率的数码相机和拍照注射胎盘。确保，拍摄照片的胎盘垂直。 </li></ol> 5。代表性的成果： 在单绒毛膜双胎的胎盘血管架构根据不同的类型单绒毛膜双胎妊娠。注射液的研究表明，AA，AV和VV吻合目前分别为80％，95％和20％，无并发症的单绒毛膜双胎妊娠1（图1 ）。机管局吻合被认为是防止TTTS和水龙头6的发展。注射液的研究表明，机管局吻合发生TTTS的胎盘中只有20％和10％的水龙头胎盘 1,2,7（图2） 。 TTTS的胎盘在激光手术fetoscopic治疗，激光血管吻合的凝血造成的伤痕可以看出，胎盘表面（图3和4）。丝锥胎盘的特点是只有几个微不足道的AV吻合（图5）的存在。胎盘的TAPS收件人的份额往往是过多的，而捐助的胎盘份额是苍白无力的（图6）。单绒毛膜双胎妊娠与出生体重不协调，生长受限胎儿往往有一个velamentous线插入和胎盘份额小得多（图7）。机管局吻合往往是单绒毛膜胎盘双胞胎出生体重不协调8。单绒毛膜monoamniotic胎盘有一个特点血管结构与短的距离，两线插入（图8）。 monoamniotic单绒毛膜胎盘机管局吻合的发病率几乎是100％， 并防止在9对双胞胎monoamniotic发展的TTTS。在这篇文章中的数字显示，色粉中心注入的单绒毛膜胎盘的特征结果。我们经常使用较深的颜色（蓝色或绿色）为动脉和浅色脉（黄色，粉红色或橙色）。 <img alt="图1" src="/files/ftp_upload/3208/3208fig1.jpg" /> 图1从正常的，简单的单绒毛膜双显示几个影音绿色动脉吻合粉色静脉（白星），从蓝动脉到黄，脉（绿星）和1个大AA吻合（由确定的几个VA吻合怀孕单绒毛膜胎盘 。的染料混合蓝色和绿色，蓝色星）。 <img alt="图2" src="/files/ftp_upload/3208/3208fig2.jpg" /> 图2显示唯一的AV（白星）和VA存在与串行amnioreduction治疗TTTS怀孕的单绒毛膜胎盘（绿星）没有AA吻合吻合。 <img alt="图3" src="/files/ftp_upload/3208/3208fig3.jpg" /> 图3。TTTS使用选择性激光技术，在其中一个小的残余吻合无意中留下的专利（白星）的血管吻合后fetoscopic激光凝固胎盘。选择性激光技术，血管吻合是首次发现，随后凝固逐一。 <img alt="图4" src="/files/ftp_upload/3208/3208fig4.jpg" /> 图4。TTTS胎盘后使用所罗门技术在其中，每一个人的吻合后，识别和凝血，完整的血管赤道是从一个胎盘保证金凝固其他fetoscopic激光凝固。 <img alt="图5" src="/files/ftp_upload/3208/3208fig5.jpg" /> 图5。TAPS的胎盘，其中只有少数，微不足道的弗吉尼亚州吻合（白星）沿血管赤道可见。 <img alt="图6" src="/files/ftp_upload/3208/3208fig6.jpg" /> 图6。产妇胎盘的TAPS（如图5所示），表明双方胎盘股的特征色差。 <img alt="图7" src="/files/ftp_upload/3208/3208fig7.jpg" /> 图7单绒毛膜双胎妊娠与双胞胎中的一个选择性宫内生长受限胎盘。生长受限胎儿有velamentous线插入和胎盘份额小得多。白星表示AA吻合。 <img alt="图8" src="/files/ftp_upload/3208/3208fig8.jpg" /> 图8。Monoamniotic胎盘：注意各种AV（绿星），VA（蓝星）和2节AA吻合（白星），和两线插入短的距离。

<ol>
	<li>Lopriore, E., Middeldorp, J. M., Sueters, M., Vandenbussche, F. P., Walther, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twin-to-twin+transfusion+syndrome:+from+placental+anastomoses+to+long-term+neurodevelopmental+outcome.">Twin-to-twin transfusion syndrome: from placental anastomoses to long-term neurodevelopmental outcome.</a> Curr. Pediatr. Rev. 1, 191-203 (2005).</li><li>Slaghekke, F., Kist, W. J., Oepkes, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twin+anemia-polycythemia+sequence:+diagnostic+criteria,+classification,+perinatal+management+and+outcome.">Twin anemia-polycythemia sequence: diagnostic criteria, classification, perinatal management and outcome.</a> Fetal Diagn. Ther. 27, 181-190 (2010).</li><li>Lopriore, E., Middeldorp, J. M., Oepkes, D., Klumper, F. J., Walther, F. J., Vandenbussche, F. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Residual+anastomoses+after+fetoscopic+laser+surgery+in+twin-to-twin+transfusion+syndrome:+frequency,+associated+risks+and+outcome.">Residual anastomoses after fetoscopic laser surgery in twin-to-twin transfusion syndrome: frequency, associated risks and outcome.</a> Placenta. 28, 204-208 (2007).</li><li>Lopriore, E., Slaghekke, F., Middeldorp, J. M., Klumper, F. J., Oepkes, D., Vandenbussche, F. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Residual+anastomoses+in+twin-to-twin+transfusion+syndrome+treated+with+selective+fetoscopic+laser+surgery:+localization,+size,+and+consequences.">Residual anastomoses in twin-to-twin transfusion syndrome treated with selective fetoscopic laser surgery: localization, size, and consequences.</a> Am. J. Obstet. Gynecol. , (2009).</li><li>Lewi, L., Jani, J., Cannie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intertwin+anastomoses+in+monochorionic+placentas+after+fetoscopic+laser+coagulation+for+twin-to-twin+transfusion+syndrome:+is+there+more+than+meets+the+eye.">Intertwin anastomoses in monochorionic placentas after fetoscopic laser coagulation for twin-to-twin transfusion syndrome: is there more than meets the eye.</a> Am. J. Obstet. Gynecol. 194, 790-795 (2006).</li><li>Denbow, M. L., Cox, P., Taylor, M., Hammal, D. M., Fisk, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+angioarchitecture+in+monochorionic+twin+pregnancies:+relationship+to+fetal+growth,+fetofetal+transfusion+syndrome,+and+pregnancy+outcome.">Placental angioarchitecture in monochorionic twin pregnancies: relationship to fetal growth, fetofetal transfusion syndrome, and pregnancy outcome.</a> Am. J. Obstet. Gynecol. 182, 417-426 (2000).</li><li>van Meir, H., Slaghekke, F., Lopriore, E., van Wijngaarden, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterio-arterial+anastomoses+do+not+prevent+the+development+of+twin+anemia-polycythemia+sequence.">Arterio-arterial anastomoses do not prevent the development of twin anemia-polycythemia sequence.</a> Placenta. 31, 163-165 (2010).</li><li>De Paepe, M. E., Shapiro, S., Young, L., Luks, F. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+characteristics+of+selective+birth+weight+discordance+in+diamniotic-monochorionic+twin+gestations.">Placental characteristics of selective birth weight discordance in diamniotic-monochorionic twin gestations.</a> Placenta. 31, 380-386 (2010).</li><li>Umur, A., van Gemert, M. J., Nikkels, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoamniotic-versus+diamniotic-monochorionic+twin+placentas:+anastomoses+and+twin-twin+transfusion+syndrome.">Monoamniotic-versus diamniotic-monochorionic twin placentas: anastomoses and twin-twin transfusion syndrome.</a> Am. J. Obstet. Gynecol. 189, 1325-1329 (2003).</li><li>Senat, M. V., Deprest, J., Boulvain, M., Paupe, A., Winer, N., Ville, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoscopic+laser+surgery+versus+serial+amnioreduction+for+severe+twin-to-twin+transfusion+syndrome.">Endoscopic laser surgery versus serial amnioreduction for severe twin-to-twin transfusion syndrome.</a> N. Engl. J. Med. 351, 136-144 (2004).</li><li>Lopriore, E., Sueters, M., Middeldorp, J. M., Vandenbussche, F. P., Walther, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Haemoglobin+differences+at+birth+in+monochorionic+twins+without+chronic+twin-to-twin+transfusion+syndrome.">Haemoglobin differences at birth in monochorionic twins without chronic twin-to-twin transfusion syndrome.</a> Prenat. Diagn. 25, 844-850 (2005).</li><li>Papathanasiou, D., Witlox, R., Oepkes, D., Walther, F. J., Bloemenkamp, K. W., Lopriore, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monochorionic+twins+with+ruptured+vasa+previa:+double+trouble!.">Monochorionic twins with ruptured vasa previa: double trouble!.</a> Fetal Diagn. Ther. 28, 48-50 (2010).</li><li>Weingertner, A. S., Kohler, A., Kohler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+and+placental+characteristics+in+four+new+cases+of+twin+anemia-polycythemia+sequence.">Clinical and placental characteristics in four new cases of twin anemia-polycythemia sequence.</a> Ultrasound Obstet. Gynecol. 35, 490-494 (2010).</li><li>Lewi, L., Cannie, M., Blickstein, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+sharing,+birthweight+discordance,+and+vascular+anastomoses+in+monochorionic+diamniotic+twin+placentas.">Placental sharing, birthweight discordance, and vascular anastomoses in monochorionic diamniotic twin placentas.</a> Am. J. Obstet. Gynecol. 197, 587-588 (2007).</li><li>Lewi, L., Gucciardo, L., Huber, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+outcome+and+placental+characteristics+of+monochorionic+diamniotic+twin+pairs+with+early-+and+late-onset+discordant+growth.">Clinical outcome and placental characteristics of monochorionic diamniotic twin pairs with early- and late-onset discordant growth.</a> Am. J. Obstet. Gynecol. , (2008).</li><li>De Paepe, M. E., Friedman, R. M., Poch, M., Hansen, K., Carr, S. R., Luks, F. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+findings+after+laser+ablation+of+communicating+vessels+in+twin-to-twin+transfusion+syndrome.">Placental findings after laser ablation of communicating vessels in twin-to-twin transfusion syndrome.</a> Pediatr. Dev. Pathol. 7, 159-165 (2004).</li><li>De Paepe, M. E., DeKoninck, P., Friedman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+distribution+patterns+in+monochorionic+twin+placentas.">Vascular distribution patterns in monochorionic twin placentas.</a> Placenta. 26, 471-475 (2005).</li><li>Nikkels, P. G., Hack, K. E., vanGemert, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+twin+placentas+with+special+attention+to+monochorionic+twin+placentas.">Pathology of twin placentas with special attention to monochorionic twin placentas.</a> J. Clin. Pathol. 61, 1247-1253 (2008).</li><li>Quintero, R. A., Martinez, J. M., Lopez, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+placental+territories+after+selective+laser+photocoagulation+of+communicating+vessels+in+twin-twin+transfusion+syndrome.">Individual placental territories after selective laser photocoagulation of communicating vessels in twin-twin transfusion syndrome.</a> Am. J. Obstet. Gynecol. 192, 1112-1118 (2005).</li><li>Quintero, R. A., Comas, C., Bornick, P. W., Allen, M. H., Kruger, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+versus+non-selective+laser+photocoagulation+of+placental+vessels+in+twin-to-twin+transfusion+syndrome.">Selective versus non-selective laser photocoagulation of placental vessels in twin-to-twin transfusion syndrome.</a> Ultrasound Obstet. Gynecol. 16, 230-236 (2000).</li></ol>

没有利益冲突的声明。

TTTS和水龙头都可以发生在单绒毛膜双胞胎的两个严重障碍。这两种疾病是由于造成不平衡intertwin血流AV吻合。在过去20年中，fetoscopic激光手术成为被证明是最好的方法治疗TTTS 10 。 fetoscopic激光治疗的目的是通过胎盘表面的血管吻合凝固的跨双循环中断。然而，TTTS激光治疗是远远不够完善。用激光治疗可导致多种并发症，包括宫内胎儿死亡，复发TTTS和水龙头1,4。这些并发症可以?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论</th></tr><tr><td>脐导管（各种尺寸，长度40厘米） </td><td> Vygon </td><td> 1270.08（8）1270.08（5）1270.04（4）1270.03（3.5 F）1270.02（2.5 F） </td><td>可用于任何类型的导管</td></tr><tr><td> 20毫升注射器</td><td>屋宇署Plastipak </td><td> 300613 </td><td></td></tr><tr><td>色粉（不同颜色） </td><td>皇家Talens Schoolverf </td><td> 36716010（蓝色）36715010（绿色）36712350（黄）36713570（粉红色） </td><td>可用于任何粘性色粉</td></tr></tbody></table>

accurate and simple evaluation of vascular anastomoses in monochorionic placenta using colored dye

胎盘血管吻合的存在是双对双胞胎输血症候群（TTTS）和双性贫血真性红细胞增多症序列（TAPS）1,2 的发展非必要条件。双胞胎的胎盘注射液的研究表明，这种吻合几乎都是目前在单绒毛膜双胞胎， 双胞胎 1 dichorionic极为罕见。已记录三种类型的吻合：从动脉到动脉，静脉静脉和动脉静脉。动静脉（AV）吻合是单向的，被称为“深”吻合，因为他们继续通过一个共同的胎盘子叶的，而动脉硬化，动脉（AA）和静脉 - 静脉（VV）吻合是双向的，被称为“肤浅”，因为他们的绒毛膜板的谎言。 TTTS和水龙头都引起血流由于影音吻合的双胞胎之间的净不平衡。双胞胎中的一个（捐赠者）的血液通过动脉泵到共享胎盘子叶，然后通过静脉排入其他双胞胎（收件人）的流通。除非血液泵回通过相反指示深AV吻合或通过浅表吻合，从收件人的捐助，发生的血液量不平衡，逐渐导致TTTS或水龙头的发展。机管局吻合的存在已被证明对保护补偿单向AV 吻合 1,2造成循环系统不平衡发展的TTTS和水龙头。单绒毛膜胎盘注射剂在出生后不久是一个有用的平均了解各种（血液）在单绒毛膜双胎的并发症的病因，是必需的测试，以达到诊断的水龙头2。此外，注射fetoscopic激光手术治疗的TTTS的胎盘可以识别可能残留吻合 3-5 。这些额外的信息是至关重要TTTS或水龙头单绒毛膜双胞胎的管理和护理工作的所有perinatologists的。当前正在使用的几个胎盘注射技术。我们提供了一个简单的协议，准确地评价（剩余）使用彩色染料注射血管吻合的存在。

Watch this Scientific Journal Video about Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye at JoVE.com

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

双对双胞胎输血症候群和双性贫血真性红细胞增多症序列是两个潜在的破坏性，在围产医学的问题。这两种疾病只发生在单绒毛膜双胞胎和不平衡的血流通过胎盘血管吻合的结果。我们提供了一个简单的协议，准确地评价使用彩色染料注射出生后，胎盘血管的血管吻合的存在。

在单绒毛膜胎盘血管吻合用有色染料的精确和简单的评价

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

accurate-simple-evaluation-vascular-anastomoses-monochorionic

Research

JoVE Journal

Medicine

26.6K 次观看.  Leiden University Medical Center. 双对双胞胎输血症候群和双性贫血真性红细胞增多症序列是两个潜在的破坏性，在围产医学的问题。这两种疾病只发生在单绒毛膜双胞胎和不平衡的血流通过胎盘血管吻合的结果。我们提供了一个简单的协议，准确地评价使用彩色染料注射出生后，胎盘血管的血管吻合的存在。

视频: Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&amp;#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

使用全血刺激试验的促炎性细胞因子IL -1β的精确和简单的测量

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

科研

医学

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

导丝辅助导管和彩色染料注射的双人单绒毛膜胎盘血管映射

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

一个简单的方法小鼠肺插管

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Fast and Accurate Exhaled Breath Ammonia Measurement

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that uses a quantum cascade based laser. The monitor is coupled to a sampler that measures mouth pressure and carbon dioxide. The system is temperature controlled and specifically designed to address the reactivity of this compound. The sampler provides immediate feedback to the subject and the technician on the quality of the breath effort. Together with the quick response time of the monitor, this system is capable of accurately measuring exhaled breath ammonia representative of deep lung systemic levels. 
Because the system is easy to use and produces real time results, it has enabled experiments to identify factors that influence measurements. For example, mouth rinse and oral pH reproducibly and significantly affect results and therefore must be controlled. Temperature and mode of breathing are other examples. As our understanding of these factors evolves, error is reduced, and clinical studies become more meaningful. This system is very reliable and individual measurements are inexpensive. 
The sampler is relatively inexpensive and quite portable, but the monitor is neither. This limits options for some clinical studies and provides rational for future innovations.

Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

快速，准确的呼出气测量氨

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or subvert anti-tumor immune responses in order to survive and flourish. Tumors take advantage of a number of different mechanisms of immune &#8220;escape,&#8221; including the recruitment of tolerogenic DC, immunosuppressive regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSC) that inhibit cytotoxic anti-tumor responses. Conversely, anti-tumor effector immune cells can slow the growth and expansion of malignancies: immunostimulatory dendritic cells, natural killer cells which harbor innate anti-tumor immunity, and cytotoxic T cells all can participate in tumor suppression. The balance between pro- and anti-tumor leukocytes ultimately determines the behavior and fate of transformed cells; a multitude of human clinical studies have borne this out. Thus, detailed analysis of leukocyte subsets within the tumor microenvironment has become increasingly important. Here, we describe a method for analyzing infiltrating leukocyte subsets present in the tumor microenvironment in a mouse tumor model. Mouse B16 melanoma tumor cells were inoculated subcutaneously in C57BL/6 mice. At a specified time, tumors and surrounding skin were resected en bloc and processed into single cell suspensions, which were then stained for multi-color flow cytometry. Using a variety of leukocyte subset markers, we were able to compare the relative percentages of infiltrating leukocyte subsets between control and chemerin-expressing tumors. Investigators may use such a tool to study the immune presence in the tumor microenvironment and when combined with traditional caliper size measurements of tumor growth, will potentially allow them to elucidate the impact of changes in immune composition on tumor growth. Such a technique can be applied to any tumor model in which the tumor and its microenvironment can be resected and processed.

This protocol describes a method for the detailed evaluation of leukocyte subsets within the tumor microenvironment in a mouse tumor model. Chemerin-expressing B16 melanoma cells were implanted subcutaneously into syngeneic mice. Cells from the tumor microenvironment were then stained and analyzed by flow cytometry, allowing for detailed leukocyte subset analyses.

在皮下肿瘤模型的评估肿瘤浸润白细胞亚群

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

套筒技术在主动脉移植小鼠模型中的应用-教学视频

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Model Surgical Training: Skills Acquisition in Fetoscopic Laser Photocoagulation of Monochorionic Diamniotic Twin Placenta Using Realistic Simulators

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome (TTTS), but is technically challenging and can lead to significant complications. Acquiring and maintaining the necessary surgical skills require consistent practice, a critical caseload, and time. Training on realistic surgical simulators can potentially shorten this steep learning curve and enables several proceduralists to acquire procedure-specific skills simultaneously. Here we describe realistic simulators designed to allow the user familiarity with the equipment and specific steps required in the surgical treatment of TTTS, including fetoscopic handling, approaches to anterior and posterior placenta, recognition of anastomoses, and efficient coagulation of vessels. We describe the skills that are especially important in conducting placental laser coagulation that the surgeon can practice on the model and apply in a clinical case. These models can be adapted easily depending on the availability of materials and require standard fetoscopy equipment. Such training systems are complementary to traditional surgical apprenticeships and can be useful aids for fetal medicine units that provide this clinical service.

Practicing the specific skills required for fetoscopic laser coagulation of monochorionic placental anastomoses on realistic models can aid less experienced surgeons in overcoming the steep learning curve associated with this procedure that is now regarded as the standard of care for twin-twin transfusion syndrome.

模型外科训练: 用逼真模拟器 Fetoscopic 激光光凝单绒毛膜 Diamniotic 双胎胎盘的技能习得

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome ...

Noninvasive Determination of Vortex Formation Time Using Transesophageal Echocardiography During Cardiac Surgery

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular (LV) filling compared with a continuous linear jet. Vortex ring development is most often quantified with vortex formation time (VFT), a dimensionless parameter based on fluid ejection from a rigid tube. Our group is interested in factors that affect LV filling efficiency during cardiac surgery. In this report, we describe how to use standard two-dimensional (2D) and Doppler transesophageal echocardiography (TEE) to noninvasively derive the variables needed to calculate VFT. We calculate atrial filling fraction (&#946;) from velocity-time integrals of trans-mitral early LV filling and atrial systole blood flow velocity waveforms measured in the mid-esophageal four-chamber TEE view. Stroke volume (SV) is calculated as the product of the diameter of the LV outflow track measured in the mid-esophageal long axis TEE view and the velocity-time integral of blood flow through the outflow track determined in the deep transgastric view using pulse-wave Doppler. Finally, mitral valve diameter (D) is determined as the average of major and minor axis lengths measured in orthogonal mid-esophageal bicommissural and long axis imaging planes, respectively. VFT is then calculated as 4 &#215; (1-&#946;) &#215; SV/(&#960;D3). We have used this technique to analyze VFT in several groups of patients with differing cardiac abnormalities. We discuss our application of this technique and its potential limitations and also review our results to date. Noninvasive measurement of VFT using TEE is straightforward in anesthetized patients undergoing cardiac surgery. The technique may allow cardiac anesthesiologists and surgeons to assess the impact of pathological conditions and surgical interventions on LV filling efficiency in real time.

We describe a protocol to measure vortex formation time, an index of left ventricular filling efficiency, using standard transesophageal echocardiography techniques in patients undergoing cardiac surgery. We apply this technique to analyze vortex formation time in several groups of patients with differing cardiac pathologies.

经食管超声心动图在心脏手术中无创地测定涡旋形成时间

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular ...

Echocardiographic Evaluation of Atrial Communications before Transcatheter Closure

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale (PFO) detection, for patient selection for transcatheter ASD/PFO closure,&#160;for intraoperative guidance and for long-term follow-up. The size, shape, location and the number of the atrial communications schould be determined. The accuracy of PFO detection can be improved by using agitated saline together with maneuvers to transiently increase the right atrial (RA) pressure. The appearance of microbubbles in the left atrium (LA) within 3 cardiac cycles after opacification of the RA is considered positive for the presence of an intracardiac shunt. Three dimensional TEE identifies further septal fenestrations and describes the dynamic morphology of ASD/PFO and atrial septal aneurysm. Follow-up evaluations with TTE is recommended at 1, 6, and 12 months after the procedure, with a subsequent evaluation every year. Previous studies showed an increased incidence of atrial arrhythmias early after device closure. Speckle tracking analysis may help to understand functional left atrial remodeling following percutaneous closure and its impact on atrial arrhythmias.

Transthoracic (TTE) and transesophageal (TEE) echocardiography represent the basic imaging tools for interatrial septum examination. Three dimensional (3D) TEE provides incremental information in the assessment of the interatrial septum. Further advanced echocardiography techniques using speckle tracking echocardiography are applied for sensitive volumetric and functional assessment of the heart chambers.&#160;

经导管闭合前心房通路的超声心动图评估

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale ...