由国立卫生研究院（NIH）的补助金，以R01-HL088424提供的资金，退伍军人事务部优异奖以聚吡咯。

1。海绵的制备<ol><li>由0.9％（W / V）氯化钠水溶液中搅拌过夜水合物海绵。 </li><li>消毒灭菌他们水合海绵。对于75毫升，高压灭菌25分钟121°C。 </li></ol>注:载入海绵具有如下所述的治疗是可选的。 <ol start="3"><li>在无菌罩与不育的仪器，从解决方案使用镊子拿起一个海绵。镊子挤压海绵和使用真空尖，尽可能消除尽可能多的解决方案，从海绵。在组织培养皿中放置了海绵拧干，小心离开海绵之间的空间。多个孔板，可用于区分不同的治疗方法。 </li><li>直接到中心的海绵吸取的处理方案。海绵轻轻按下枪头后的解决方案已被放置在海绵上，这有助于海绵吸收的解决方案。注:6毫米直径2.75毫米厚的海绵，我们聘请的是能够容纳最多25μL。 </li><li>板，他们在所有的海绵一旦被加载，可以存储在冰块，直到他们在植入老鼠。 </li></ol> 2。外科手术<ol><li>适当麻醉动物和管理先发制人镇痛。整个过程将持续约20分钟，每头牲畜。对于30Ğ鼠标每分钟氧气的1.5升与1.5％异氟醚用于诱导和维持麻醉。当动物不再回应捏反射试验，在仰卧位在鼻锥的地方。使用电动剃须刀刮胡子1英寸1英寸面积上的低左肋下腹部。擦拭干净，剪去长发。 </li><li>剃光面积的70％的酒精优碘清洗消毒。重复此优碘酒精清洗三次。 </li><li>教而持有的皮肤，用手术刀作垂直切口直径的1.5-2倍要插入海绵米，是谨慎的，并非要削减通过体腔。 </li><li>使用镊子，以保持皮肤的边缘，并慢慢开始使用Metzembaum剪刀切口两侧皮肤从肌肉中分离出来。口袋应该整个鼠标的宽度，从肋骨到后腿。 </li><li>使用镊子，拿起第一个被放置在动物从两侧捏住海绵。海绵转移到直镊子，因此，它是从顶部和底部举行。 </li><li>使用镊子，以保持皮肤的边缘和解除它，这样可以插入海绵。慢慢地插入海绵，尽量避免接触皮肤或肌肉下方的海绵将难以移动，一旦触及组织。重复所有的海绵。 </li><li>一旦所有的海绵已经插入，地方两个或三个缝合关闭切口。使用一对镊子一起举行切口两侧和线程的苏图再通过两个层次的皮肤。拧紧第一个罚球刚够关闭皮肤边缘。完成一个从另一个方向，这个时候拉结紧罚的平结。然后把两个多平方海里，完成缝合（ 图2）。放置额外的缝合关闭整个切口。其他伤口粘接方法可用于如dermabond，伤口剪辑，或订书钉。 </li><li>从动物麻醉系统和监控，而意识恢复。动物应尽快开始走动，并返回到前运算流动。确保他们能够充分达到食物和水。有些湿润的食物，可放置在笼，以协助恢复的底部。继续每天监测动物遇险或不适的迹象，检查切口部位的感染，炎症，和裂开。如果发现任何复苏的并发症，采取适当的行动。 </li></ol> 3。可选注射液<ol><li>海绵注入所需的解决方案，首先要确定管理鼠标的物质的量。该物质需要集中，使注射量是不到一半的海绵可容纳的总体积。 </li><li>准备一个注射器注射（我们用28Ğ针的的小鼠）。 </li><li>将动物麻醉下接受注射。 </li><li>当动物不再响应捏测试可以注射。 </li><li>为了给推注射针，在45°的角度深入皮肤海绵中心。只去了一半的方式，然后推入针的海绵通过它的厚度。我们的目标是注入海绵确切中心。 </li><li>进行检查，以确保针是海绵，慢慢抬起垂直针。如果针是海绵，海绵应该向上移动的针。 </li><li>慢慢注入海绵取出针。如果有多个需要注射海绵很可能将被推迟，一些注射液注射前网站。 </li></ol> 4。海绵去除注意:在海绵去除，处理时要格外小心海绵。避免穿透的海绵，手术器械和海绵，避免对周围组织周围的主要动脉防止失血过多。 <ol><li>动物是安乐死后，去除皮肤使用镊子和切口关闭海绵的海绵。 </li><li>仔细分离的海绵，从周围的胶囊<html> ND避免收获额外的组织周围的海绵。如给药或形态分析，精度较差，需要切除。 </li></ol> 5。代表结果删除海绵可以存储在不同的条件取决于将要执行的分析类型。对于切片，撷取海绵可以嵌入在一个中等的最佳切削温度（OCT）的化合物，如冻结或放置在10％福尔马林嵌入切片和石蜡块（ 图3A和B）。切片的最好结果，得到海绵时减少了一半整个直径和嵌入式切面（ 图4A和B）。应采取的海绵部分，使整个海绵宽度图3A显示了两个海绵，海绵左嵌入式/切片不正确，正确处理和正确的海绵。 耳鼻喉科"&gt;海绵进行分析，可以放置在一个Eppendorf管中，并储存在-20°C，直到准备处理的RNA和实时定量PCR分析海绵应放置在一个Eppendorf管，闪光冻结，并存储在 - 80°C。此外，如从凯杰RNAlater RNA的防腐剂，可以稳定在较高温度下的安全储存的RNA。创面渗液多管挤海绵，可以收集，并汇集了流体多个海绵，以获得具有代表性的样本。活细胞，如成纤维细胞，巨噬细胞和淋巴细胞，也可以从海绵中提取，去除后从动物，海绵，可以在适当的媒体所需的细胞类型的地方。海绵可以处理物理（即切碎）或酶（如胶原酶）方法以释放细胞。 <img alt="图1。" src="/files/ftp_upload/3885/3885fig1.jpg" /> 图1。脱水特德PVA海绵在三种不同尺寸的磁盘。 <img alt="图2。" src="/files/ftp_upload/3885/3885fig2.jpg" /> 图2小鼠的伤口愈合模型显示侧切伤口的位置和四个聚乙烯醇海绵的图片。 <img alt="图3。" src="/files/ftp_upload/3885/3885fig3.jpg" /> 图3。 （一）在4X H＆E染色和（B）三色染色切片石蜡嵌入海绵10X。 <img alt="图4。" src="/files/ftp_upload/3885/3885fig4.jpg" /> 图4。 （一）在沿直径的一半海绵切割图片（B）PVA海绵一半，嵌入式削减10月朝下。

<ol>
	<li>Efron, D. T., Barbul, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subcutaneous+sponge+models.">Subcutaneous sponge models.</a> Methods Mol. Med. 78, 83-93 (2003).</li><li>Gottrup, F., Agren, M. S., Karlsmark, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+use+in+wound+healing+research:+a+survey+focusing+on+in+vitro+and+in+vivo+adult+soft+tissue.">Models for use in wound healing research: a survey focusing on in vitro and in vivo adult soft tissue.</a> Wound Repair. 8, 83-96 (2000).</li><li>Sprugel, K. H., McPherson, J. M., Clowes, A. W., Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+growth+factors+in+vivo.+I.+Cell+ingrowth+into+porous+subcutaneous+chambers.">Effects of growth factors in vivo. I. Cell ingrowth into porous subcutaneous chambers.</a> Am. J. Pathol. 129, 601-613 (1987).</li><li>Alaish, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+polyvinyl+alcohol+sponge+and+expanded+polytetrafluoroethylene+subcutaneous+implants+as+models+to+evaluate+wound+healing+potential+in+human+beings.">Comparison of the polyvinyl alcohol sponge and expanded polytetrafluoroethylene subcutaneous implants as models to evaluate wound healing potential in human beings.</a> Wound Repair. 3, 292-298 (1995).</li><li>Davidson, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+wound+repair.">Animal models for wound repair.</a> Arch. Dermatol. Res. 290, 1-11 (1998).</li><li>Alfaro, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=sFRP2+suppression+of+bone+morphogenic+protein+(BMP)+and+Wnt+signaling+mediates+mesenchymal+stem+cell+(MSC)+self-renewal+promoting+engraftment+and+myocardial+repair.">sFRP2 suppression of bone morphogenic protein (BMP) and Wnt signaling mediates mesenchymal stem cell (MSC) self-renewal promoting engraftment and myocardial repair.</a> J. Biol. Chem. 285, 35645-35653 (2010).</li><li>Andrade, S. P., Ferreira, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+sponge+implant+model+of+angiogenesis.">The sponge implant model of angiogenesis.</a> Methods Mol. Biol. 467, 295-304 (2009).</li><li>Diegelmann, R. F., Lindblad, W. J., Cohen, I. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+subcutaneous+implant+for+wound+healing+studies+in+humans.">A subcutaneous implant for wound healing studies in humans.</a> J. Surg. Res. 40, 229-237 (1986).</li><li>Efron, D. T., Most, D., Shi, H. P., Tantry, U. S., Barbul, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+of+studying+wound+healing.">A novel method of studying wound healing.</a> J. Surg. Res. 98, 16-20 (2001).</li><li>Lindblad, 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=Considerations+for+selecting+the+correct+animal+model+for+dermal+wound-healing+studies.">Considerations for selecting the correct animal model for dermal wound-healing studies.</a> J. Biomater. Sci. Polym. Ed. 19, 1087-1096 (2008).</li></ol>

我们什么都没有透露。

<p class="jove_content"&gt;伤口愈合海绵模型有多种变化，它可以在许多不同的物种，包括人类和海绵进行，也可植入背部<sup&gt; 1,2,8</sup&gt;。对于人类和其他物种的研究，海绵可能需要放置在多孔硅管，海绵，以防止被封装并允许便于搬迁<sup&gt; 4,8</sup&gt;。海绵植入的位置，可以改变预期的效果和技师技能的基础上。背植入可以让海绵在皮肤下移动，而腹侧植入都比较困难由于切割腹腔概率。海绵可以设置用导管给药，使常数和控制，消除了需要多次注射<sup&gt; 9</sup&gt;。甚至可以通过针注射较小，管状海绵<sup&gt; 8</sup&gt;。不仅限于只有聚乙烯醇海绵的组成。他们可以准备许多材料完全或合并<sup&gt; 7</sup&gt;。</p&gt;<p class="jove_content"&gt;这是海绵准备和动物的位置是一致的，正确的理解是至关重要的。如果从表削减海绵，它们必须是类似的可比性重量。如果海绵细胞或治疗是摆在每个海绵金额必须是准确的。愈合过程中可以改变的基础上安置在动物和愈合海绵的位置，从头部的增加将减少距离<sup&gt; 1,10</sup&gt;。</p

<table border="1"><tbody><tr><td> 材料名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> PVA海绵</td><td>美敦力公司</td><td> CF120 </td><td>的大小和海绵的孔隙率取决于实验</td></tr><tr><td>手术刀刀片大小15 </td><td>屋宇署内侧</td><td> 371115 </td><td></td></tr><tr><td> metzembaum剪刀</td><td> Thermo Scientific的</td><td> 79-211 </td><td></td></tr><tr><td>止血钳</td><td>罚款科学工具</td><td> 13004-14 </td><td></td></tr><tr><td>持针器</td><td>罚款科学工具</td><td> 12004-16 </td><td></td></tr><tr><td> 5-0尼龙线</td><td> ethilon </td><td> 698 </td><td></td></tr><tr><td>华侨城compouND </td><td>组织TEK </td><td> 4583 </td><td></td></tr></tbody></table>

the polyvinyl alcohol sponge model implantation

伤口愈合是一个复杂的，多步骤的过程，涉及到许多类型的细胞，生长因子和化合物1-3。由于这种复杂性，促进伤口愈合的研究时，在体内进行的最全面的。有许多在体内模型可用来研究急性伤口愈合，包括切口，切除，死亡空间，烧伤。死亡空间模型是人为的，这是用来研究组织的形成和物质的影响，在伤口上的多孔植入物。一些常用的死空间模型包括聚乙烯醇（PVA）海绵，钢丝网气瓶，膨体聚四氟乙烯（膨体聚四氟乙烯）材料，与Cellstick 1,2。 每个死区空间模型有其自身的局限性，根据其材料的成分和植入方法。钢丝网圆柱模型有一个植入后浸润的滞后期，需要相当长的时间前肉芽组织形成begiNS 1。后来伤口愈合的阶段是最好的分析使用的聚四氟乙烯模式1,4。在该Cellstick是硅管模式，它通常被用于研究人类的手术伤口，伤口液2内的纤维素海绵。 PVA海绵急性研究是有限的，因为随着时间的推移，它开始引起异物反应，导致巨细胞反应，在动物5。不像其他材料，聚乙烯醇海绵很容易插入和删除，惰性和非生物降解材料制成的，但有足够的软，切片的病理分析2,5。 在伤口愈合的PVA海绵是非常有用的分析肉芽组织形成，胶原沉积，伤口液成分，物质1,2,5愈合过程的影响。除了利用其在学习各种各样的伤口愈合的属性，PVA海绵也被用在许多其他类型的研究。它Ĥ作为被用来研究肿瘤血管生成的药物输送和干细胞的存活和植入1,2,6,7。 PVA海绵随着伟大更改性，事先广泛使用，和可重复性的结果，是1,2许多研究的理想模型。 在这里，我们将描述的准备，植入和检索PVA海绵磁盘，在伤口愈合的小鼠模型（ 图1）。

Watch this Scientific Journal Video about The Polyvinyl Alcohol Sponge Model Implantation at JoVE.com

The Polyvinyl Alcohol Sponge Model Implantation

一个有用的工具，药物，生长因子，和/或操纵细胞在创面修复的动物模型的影响分析描述。这种技术利用了聚乙烯醇（PVA）海绵的性能，提供包含所需的治疗，还提供一个被切除和分析平台。

聚乙烯醇海绵模型植入

Wound healing is a complicated, multistep process involving many cell types, growth factors and compounds1-3. Because of this complexity, wound healing ...

the-polyvinyl-alcohol-sponge-model-implantation

Research

JoVE Journal

Medicine

19.7K 次观看.  Vanderbilt University School of Medicine. 一个有用的工具，药物，生长因子，和/或操纵细胞在创面修复的动物模型的影响分析描述。这种技术利用了聚乙烯醇（PVA）海绵的性能，提供包含所需的治疗，还提供一个被切除和分析平台。

视频: The Polyvinyl Alcohol Sponge Model Implantation

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

科研

医学

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Implantation of the Syncardia Total Artificial Heart

With advances in technology, the use of mechanical circulatory support devices for end stage heart failure has rapidly increased. The vast majority of such patients are generally well served by left ventricular assist devices (LVADs). However, a subset of patients with late stage biventricular failure or other significant anatomic lesions are not adequately treated by isolated left ventricular mechanical support. Examples of concomitant cardiac pathology that may be better treated by resection and TAH replacement includes: post infarction ventricular septal defect,&#160;aortic root aneurysm / dissection, cardiac allograft failure, massive ventricular thrombus, refractory malignant arrhythmias (independent of filling pressures), hypertrophic / restrictive cardiomyopathy, and complex congenital heart disease. Patients often present with cardiogenic shock and multi system organ dysfunction. Excision of both ventricles and orthotopic replacement with a total artificial heart (TAH) is an effective, albeit extreme, therapy for rapid restoration of blood flow and resuscitation. Perioperative management is focused on end organ resuscitation and physical rehabilitation. In addition to the usual concerns of infection, bleeding, and thromboembolism common to all mechanically supported patients, TAH patients face unique risks with regard to renal failure and anemia. Supplementation of the abrupt decrease in brain natriuretic peptide following ventriculectomy appears to have protective renal effects. Anemia following TAH implantation can be profound and persistent. Nonetheless, the anemia is generally well tolerated and transfusion are limited to avoid HLA sensitization. Until recently, TAH patients were confined as inpatients tethered to a 500 lb pneumatic console driver. Recent introduction of a backpack sized portable driver (currently under clinical trial) has enabled patients to be discharged home and even return to work. Despite the profound presentation of these sick patients, there is a 79-87% success in bridge to transplantation.

The purpose of this manuscript is to briefly review indications, management, and outcomes for the total artificial heart. Video operative techniques for device implantation are presented.

SynCardia全人工心脏植入术

With advances in technology, the use of mechanical circulatory support devices for end stage heart failure has rapidly increased. The vast majority of ...

Implantation of Total Artificial Heart in Congenital Heart Disease

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital heart disease (CHD), the malformed heart presents a challenge to TAH implantation. 
In the case presented here, a 17 year-old patient with congenital transposition of the great arteries (CCTGA) experienced progressively worsening HF due to his congenital condition. He was hospitalized multiple times and received an implantable cardioverter defibrillator (ICD). However, his condition soon deteriorated to end-stage HF with multisystem organ failure. 
Due to the patient's grave clinical condition and the presence of complex cardiac lesions, the decision was made to proceed with a TAH. The abnormal arrangement of the patient's ventricles and great arteries required modifications to the TAH during implantation.
With the TAH in place, the patient was able to return home and regain strength and physical well-being while awaiting a donor heart. He was successfully bridged to heart transplantation 5 months after receiving the device. This report highlights the TAH is feasible even in patients with structurally abnormal hearts, with technical modification.

This is a case report of a patient with congenitally corrected transposition of the great arteries (CCTGA) who received a total artificial heart (TAH) as a bridge to heart transplant. The TAH was successfully implanted with modifications to accommodate the patient's congenitally malformed heart.

的全人工心脏的先天性心脏病植入

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

下腔静脉间置移植物在小鼠模型中植入

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model

The enormous upsurge of interest in immune-based treatments for cancer such as vaccines and immune checkpoint inhibitors, and increased understanding of the role of the tumor microenvironment in treatment response, collectively point to the need for immune-competent orthotopic models for pre-clinical testing of these new therapies. This paper demonstrates how to establish an orthotopic immune-competent rat model of pleural malignant mesothelioma. Monitoring disease progression in orthotopic models is confounded by the internal location of the tumors. To longitudinally monitor disease progression and its effect on circulating immune cells in this and other rat models of cancer, a single tube flow cytometry assay requiring only 25 &#181;l whole blood is described. This provides accurate quantification of seven immune parameters: total lymphocytes, monocytes and neutrophils, as well as the T-cell subsets CD4 and CD8, B-cells and Natural Killer cells. Different subsets of these parameters are useful in different circumstances and models, with the neutrophil to lymphocyte ratio having the greatest utility for monitoring disease progression in the mesothelioma model. Analyzing circulating immune cell levels using this single tube method may also assist in monitoring the response to immune-based treatments and understanding the underlying mechanisms leading to success or failure of treatment.

The generation of an orthotopic rat model of pleural malignant mesothelioma by implantation of II-45 mesothelioma cells into the pleural cavity of immune competent rats is presented. A flow cytometric method to analyze seven immune cell subsets in these animals from a 25 &#181;l&#160;blood sample is also described.

在II-45同基因大鼠模型间皮瘤原位移植和外周免疫细胞监视

The enormous upsurge of interest in immune-based treatments for cancer such as vaccines and immune checkpoint inhibitors, and increased understanding of ...

Surgical Training for the Implantation of Neocortical Microelectrode Arrays Using a Formaldehyde-fixed Human Cadaver Model

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. Recent technological progress has enabled the fabrication of microelectrode arrays that allow recording the activity of multiple individual neurons in the neocortex of the human brain. These arrays have the potential to bring unique insight onto the neuronal correlates of cerebral function in health and disease. Furthermore, the identification and decoding of volitional neuronal activity opens the possibility to establish brain-computer interfaces, and thus might help restore lost neurological functions. The implantation of neocortical microelectrode arrays is an invasive procedure requiring a supra-centimetric craniotomy and the exposure of the cortical surface; thus, the procedure must be performed by an adequately trained neurosurgeon. In order to provide an opportunity for surgical training, we designed a procedure based on a human cadaver model. The use of a formaldehyde-fixed human cadaver bypasses the practical, ethical and financial difficulties of surgical practice on animals (especially non-human primates) while preserving the macroscopic structure of the head, skull, meninges and cerebral surface and allowing realistic, operating room-like positioning and instrumentation. Furthermore, the use of a human cadaver is closer to clinical daily practice than any non-human model. The major drawbacks of the cadaveric simulation are the absence of cerebral pulsation and of blood and cerebrospinal fluid circulation. We suggest that a formaldehyde-fixed human cadaver model is an adequate, practical and cost-effective approach to ensure proper surgical training before implanting microelectrode arrays in the living human neocortex.

We designed a procedure in which a formaldehyde-fixed human cadaver is used to assist neurosurgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain.

应用甲醛固定人尸体模型植入皮层微电极阵列的外科训练

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. ...

Cochlear Implantation in the Guinea Pig

Cochlear implants are highly efficient devices that can restore hearing in subjects with profound hearing loss. Due to improved speech perception outcomes, candidacy criteria have been expanded over the last few decades. This includes patients with substantial residual hearing that benefit from electrical and acoustical stimulation of the same ear, which makes hearing preservation during cochlear implantation an important issue. Electrode impedances and the related issue of energy consumption is another major research field, as progress in this area could pave the way for fully implantable auditory prostheses. To address these issues in a systematic way, adequate animal models are essential. Therefore, the goal of this protocol is to provide an animal model of cochlear implantation, which can be used to address various research questions. Due to its large tympanic bulla, which allows easy surgical access to the inner ear, as well as its hearing range which is relatively similar to the hearing range of humans, the guinea pig is a commonly used species in auditory research. Cochlear implantation in the guinea pig is performed via a retroauricular approach. Through the bullostomy a cochleostomy is drilled and the cochlear implant electrode is inserted into the scala tympani. This electrode can then be used for electrical stimulation, determination of electrode impedances and the measurement of compound action potentials of the auditory nerve. In addition to these applications, cochlear implant electrodes can also be used as drug delivery devices, if a topical delivery of pharmaceutical agents to the cells or fluids of the inner ear is intended.

The goal of this protocol is to provide an animal model of cochlear implantation, which can be used to address a multitude of research questions. Potential applications include the evaluation of pharmaceutical interventions or electrical stimulation for beneficial effects on hearing thresholds or electrode impedances.

豚鼠耳蜗植入术

Cochlear implants are highly efficient devices that can restore hearing in subjects with profound hearing loss. Due to improved speech perception ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

The Lower Body Positive Pressure Treadmill for Knee Osteoarthritis Rehabilitation

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

Here, based on a clinician&#8217;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

下体正压跑步机用于膝关节骨关节炎康复

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in ...