This study was supported by a grant from the Dutch Kidney Foundation (KJPB 08.0003). 
Carolien Rothuizen is acknowledged for her contribution to the study. Hoang Pham is acknowledged for his assistance with the pathology work-up.

所有实验均通过在莱顿大学医学中心的动物福利委员会的批准。 1.动物的制备和麻醉<ol><li>麻醉在充满3-4％异氟醚持续麻醉诱导室鼠标（1-3个月）。 </li><li>剃颈部和使用电剃刀左大腿的内侧部分的腹侧和用一块胶带来去除毛发。 </li><li>上双眼眼应用软膏。 </li><li>在其上鼻子通气面罩固定在加热毯动物的位置。固定头部用胶带的鼻罩。 <ol><li>使用富氧空气集在下面的流在1.5-2.0％的浓度递送异氟烷:空气0.3升/分钟，100％氧气0.2升/分。捏脚趾之间的皮肤，以评估麻醉深度，并根据需要调整异氟醚的浓度。 </li></ol></li><li>固定的动物的加热毯用胶带上是在仰卧位置设定在约37℃的四肢。 </li><li>注入丁丙诺啡溶解在氯化钠（NaCl）0.9％，0.1毫克的剂量/在动物的侧翼0.5毫升无菌的0.9％生理盐水皮下注射一起公斤。 </li><li>申请洗必泰酊0.5％到准备好的区域。 </li></ol> 2.皮肤切口<ol><li>其对外科医生的头在显微镜下动物的位置。 </li><li>使在使用微剪刀腹侧颈部的中线的约1.5厘米的纵向切口。 </li><li>抓住用钳子唾液腺和颅取代正确的唾液腺，直到它部分是伤口之外。 </li></ol> 3.解剖和准备静脉<ol><li>确定并直言不讳地通过散布完全从它的血管周围组织解剖右颈外静脉的分支背内侧只是一个长边和在血管的方向。 </li><li>将一个循环（10.0缝合）周围的解剖静脉和应用一个结没有锁定它。 </li></ol> 4.删除胸锁乳突肌<ol><li>使用镊子，直言沿边境蔓延，从剖析其周围右胸锁乳突肌。将一对开放钳分离肌肉下，用6.0缝线结扎其近端和远端。随后，消费使用cauterizer肌肉。 </li></ol> 5.解剖和准备颈总动脉<ol><li>识别并使用镊子解剖右颈总动脉。放置一个6.0缝合线周围的动脉中的动脉的操作中提供帮助。 </li><li>作为远端和近端尽可能应用血管夹。 </li><li>使在约1mm使用专门微剪刀动脉的中间的纵向切口。 </li><li>冲洗动脉用肝素溶液[100 IU / ml的]，直到容器是明确的血液。 </li><li>测量并调整切口的长度，如果需要将血管夹[1.1 mm]的匹配的宽度。 </li></ol> 6.静脉结扎<ol><li>近端早已准备好的静脉应用血管钳夹，并将静脉周围缝合线。 </li><li>使用被放置在缝合线上和结扎静脉的远端尽可能使用10.0缝线被以前放置在止血剂应用温和尾部牵引到静脉。 </li><li>使用切剪刀刚好接近其结扎静脉。 </li><li>使用血管钳，轻轻打开静脉的腔，并用肝素溶液[100国际单位/毫升]冲洗。 </li></ol> 7.创建接合部1 <ol><li>静脉连接到动脉用2平方海里在十二点钟位置，随后由一个缝合在六点一个间断缝合（10.0）时钟的位置。确保将静脉动脉连接，而不让其绕其自身的轴线。为了提高手术曝光旋转动物。 </li><li>定位静脉周围缝合线，并用止血剂应用温和的横向牵引力。 </li><li>使用相同的间断缝合，通过将大约3-4额外缝线在吻合的可见腹侧完成吻合术。 </li></ol> 8.肝素管理<ol><li>旋转加热毯180°聚焦显微镜左侧大腿。 </li><li>通过寻找，在上腿的内侧部分纵向运行，并且可以通过皮肤可见血管结构识别股静脉/动脉/神经束。使用的约1cm直接在长度方式股​​静脉上方的微剪刀切开。 </li><li>小心解剖股静脉的血管周围组织和在剂量Ô注入肝素f在股静脉0.2国际单位/克体重。 </li></ol> 9.创建接合部2 <ol><li>返回到颈部区域，并删除被置于静脉周围的缝合线。 </li><li>放置缝合线（6.0），该随后通过颈动脉的下方，在静脉和再次低于颈动脉。 （ 图1L） </li><li>取下静脉血管钳。 </li><li>接着，通过同时加捻两个血管夹拧半完成的动静脉瘘沿着以顺时针方式颈动脉的轴线180°，并应用内侧牵引到缝合线的端部。 </li><li>如在步骤7.2中描述完成以同样的方式的吻合术。 </li></ol> 10.血管钳去除<ol><li>取出缝合线和旋转血管夹180°逆时针。 </li><li>移除远侧血管钳接着近端血管夹。 </li> &lt;LI&gt;轻轻阻塞与血管钳静脉流出道评估吻合的通畅。在专利吻合的情况下，预封闭的静脉部分将在一个脉动方式展开。重新定位合适的唾液腺到原来的解剖位置使用镊子。 </li></ol> 11.皮肤缝合和术后护理<ol><li>关闭上腿的皮肤与一个不间断缝合（6.0）。 </li><li>关闭颈部皮肤用不间断缝合（6.0）。 </li><li>从加热毯取出的动物，并注入0.5毫升0.9％生理盐水皮下注射。 </li><li>将动物在黑暗的笼子里是带加热灯加热，让它完全恢复。当动物没有完全恢复，确保它不从血流动力学的冲击，由于在手术区出血受苦。寻找迹象，如颈部肿胀和血液渗漏。 </li><li>后bupr的第一次注射后约6小时enorphine，注入的剂量是由制造商推荐，以便为附加的72小时提供足够的镇痛丁丙诺啡皮下的持续释放制剂的单次剂量。 </li></ol> 12.组织收获<ol><li>麻醉动物用含有咪达唑仑（5毫克/千克）腹腔内给药的麻醉剂混合物，美托咪定（0.5毫克/千克）和芬太尼（0.05毫克/千克）。 </li><li>通过插入通过其前爪和脸颊针头进入非加热的硅垫固定在仰卧位置的动物。 </li><li>使约1.5-2.0厘米的切口在与所述微剪刀颈部疤痕。 </li><li>剖析AVF，并把缝合线（6.0）围绕proximal-，远端动脉和静脉流出道，便于识别。 </li><li>作为该协议的第10.3节描述评估AVF的通畅。 </li><li>通过执行剖腹探查内侧打开ü腹腔唱微型剪刀。 <ol><li>通过使用一对在锁骨中线尾端开始剪刀的左和右腹侧胸廓切开。用微型剪刀，横切附加到腹侧肋骨的中间部分，并随后置换本条颅使用镊子隔膜。 </li><li>找到下腔静脉和用微剪横切它。 </li></ol></li><li>插入针23号针头在左心室和在约100毫米汞柱的压力下，用PBS灌注，直至血管内流体是近清楚。 </li><li>不除去针，现在用4％福尔马林在约100毫米汞柱的压力灌注10分钟。 </li><li>通过与微剪刀横切两个动脉和静脉切除AVF并在4％的福尔马林溶液O / N淹没它。 </li></ol> 13.组织包埋和剖分<ol><li>处理该组织根据制造商的protoco石蜡湖该协议包括施加如下: <ol><li>浸入乙醇70％，在室温1小时。重复2倍。 </li><li>浸入乙醇96％，在室温1小时。重复2倍。 </li><li>浸入乙醇99.5％在室温1小时。重复一次。 </li><li>浸入乙醇99.5％在室温1.5小时。 </li><li>在二甲苯浸入100％，在室温1小时。重复一次。 </li><li>在62℃浸入石蜡1小时。重复2倍。 </li></ol></li><li>嵌入AVF石蜡使得静脉流出道在垂直于包埋盒定向。 </li><li>用切片机，创建12个连续切片每一个5微米的厚度由30节。然后每个串行部分，放置在从一个位置以有序的方式开始多达十二开始最接近吻合的区域的单个安装玻璃一个部分。 </li></ol>

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作者什么都没有透露。

该AVF被认为是致命的血液透析治疗脚跟。不幸的是，AVF仍存有大量的失败8-10受到影响。尽管在底层机制的广泛研究，确切的病理生理机制仍然不明。为AVF失败众多鼠模型已在文献2,3,11,12说明。然而，这些模型的掺入静脉结束这一最在临床情况中使用动脉侧吻合配置。这是非常重要，因为将得到的血流动力学起着血管重塑中起重要作用。此外，一些模型中使用的合成的袖口的动脉?...

一个功能血管通路管道是对于依赖于长期血液透析维持生命肾功能衰竭患者至关重要。一个动静脉瘘（AVF）的建设是目前血管通路的首选。然而，AVF相关并发症构成发病的一个主要原因患者慢性血液透析。尽管进行了广泛的科学努力，没有新颖的方法，以减少AVF访问相关的并发症，并导致AVF耐用性显着改善。这一令人失望的进步的一部分涉及到血液透析访问失败的基本病理生理学的理解不够充分。 解开的AV访问失败，动物模型密切模仿人类病理学的病理生理学是极其重要的。在这方面，不但动物种类，但也吻合部位，所需的抗混凝疗法和的持续时间浪涌后跟进RY应考虑到1。虽然大型动物是最适合的目的是开发新的治疗策略的干预研究，小鼠模型已经获得在底层的AV接入失败的分子机制，更深入的了解，由于转基因小鼠的可用性的最大潜力。此外，可用于这一目的在相比更大的动物使用较低成本的大量小鼠。 AVF失败的第一鼠模型是由桂和等 2在该模型中在2004年描述的，使用的颈动脉，并使用血管内导管的端至端的方式颈静脉构建动静脉瘘。虽然端至端的配置和血管内导管的存在限制这种模型为人类动静脉瘘的有效性这种模式可能是研究在动静脉瘘的早期静脉适应有用。一种改进AVF模型由Castier和等人介绍了3，其中的端部颈动脉被连接到颈静脉的一侧。然而，在血液透析患者动静脉瘘通常由解剖静脉的端部到动脉的侧面构成。 AVF的确切配置是AV访问模型的一个重要特征，因为它决定了导管4内的血流动力学轮廓。后者是内皮功能障碍和内膜增生（IH）5的后续发展的重要因素。 一种新的小鼠模型最近被用相同的解剖形态发展为利用人类6。在这个模型中，AVF在C57BL / 6小鼠吻合颈外静脉的分支的末端间断缝合颈总动脉的一侧。在本文中，我们专注于这个模型的显微外科方法，以便于广泛使用这种小鼠模型，旨在解开复杂的病理生理机制血液透析访问失败。

<table border="0" cellpadding="0" cellspacing="0" width="697">
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:247px;">Dissecting microsocpe</td>
			<td style="width:219px;">Leica</td>
			<td style="width:117px;">M80</td>
			<td style="width:115px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forceps</td>
			<td>Medicon</td>
			<td>07.61.25</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vascular forceps</td>
			<td>S&#38;T</td>
			<td>JFL-3D.2</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vascular forceps</td>
			<td>S&#38;T</td>
			<td>D-5a.2</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forceps</td>
			<td>Roboz</td>
			<td>SS/45</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Micro scissor 5 mm blade</td>
			<td>Fine science tools</td>
			<td>15000-08</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Micro scissor 2 mm blade</td>
			<td>Fine science tools</td>
			<td>15000-03</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Scissor</td>
			<td>Medicon</td>
			<td>05.12.21</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Clip applier 1</td>
			<td>S&#38;T</td>
			<td>CAF-4</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vascular clamp 1</td>
			<td>S&#38;T</td>
			<td>B-1V</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Clip applier 2</td>
			<td>BBraun</td>
			<td>FE572K</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vascular clamp 2</td>
			<td>BBraun</td>
			<td>FE740K</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Hemostatic forceps</td>
			<td>BBraun</td>
			<td>BH110</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10.0 sutures</td>
			<td>BBraun</td>
			<td>G1117041</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6.0 sutures</td>
			<td>BBraun</td>
			<td>768464</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Cauterizer</td>
			<td>Fine science tools</td>
			<td>18010-00</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Needle holder</td>
			<td>Medicon</td>
			<td>11.82.18</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ocular ointment</td>
			<td>Pharmachemie</td>
			<td>41821101</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Chlorhexidine tincture 0,5%</td>
			<td>Leiden University Medical Center</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Heparin</td>
			<td>Leo Pharma</td>
			<td>012866-08</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Buprenorphin</td>
			<td>RB Pharmaceuticals&#160;</td>
			<td>283732</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Isoflurane</td>
			<td>Pharmachemie</td>
			<td>45,112,110</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Anesthesia mask</td>
			<td>Maastricht university</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Midazolam</td>
			<td>Actavis</td>
			<td>AAAC6877</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Dexmedetomidine</td>
			<td>Orion</td>
			<td>141-267</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Fentanyl</td>
			<td>Bipharma</td>
			<td>15923002</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Continuous anaesthetic induction chamber</td>
			<td>Vet-tech solutions</td>
			<td>AN010R</td>
			<td></td>
		</tr>
	</tbody>
</table>

a novel murine model of arteriovenous fistula failure: the surgical procedure in detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

创建吻合（图1）后，通畅应由不久包藏有血管钳静脉流出道进行评估。当吻合是专利，血管道近端闭塞应明确扩大以脉动方式。此外，通畅是通过使用近红外荧光检查（NIRF），该有效地作为一个血管造影（ 图2）确认。在手术过程的故障会导致吻合中（图2）中描绘的闭塞。这种故障可通过太窄吻合区域，容器的扭转，肝素剂量不足或吻合的前侧连接到背面?...

Watch this Scientific Journal Video about A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail at JoVE.com

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

动静脉内瘘失败的新型小鼠模型:在详细介绍手术过程

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

a-novel-murine-model-arteriovenous-fistula-failure-surgical-procedure

Research

JoVE Journal

Medicine

12.9K 次观看.  Leiden University Medical Center. Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

视频: A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and &alpha;1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual's immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.

We developed a novel surgical approach for intratracheal administration of bioactive agents into the mouse fetus. The delivery route is more efficient in targeting the fetal mouse lungs than the commonly used intra-amniotic injection. This procedure has to date not been described in a mouse model.

一种新的手术方法气管内注射生物活性剂在胎鼠模型

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

科研

医学

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

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 ...

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation of the animal for surgery, opening the thoracic body wall, exposing the desired rib from the surrounding intercostal muscles, excising the desired section of rib without inducing a pneumothorax, and closing the incisions. Compared to the bones of the appendicular skeleton, the ribs are highly accessible. In addition, no internal or external fixator is necessary since the adjacent ribs provide a natural fixation. The surgery uses commercially available supplies, is straightforward to learn, and well-tolerated by the animal. The procedure can be carried out with or without removing the surrounding periosteum, and therefore the contribution of the periosteum to repair can be assessed. Results indicate that if the periosteum is retained, robust repair occurs in 1 - 2 months. We expect that use of this protocol will stimulate research into rib repair and that the findings will facilitate the development of new ways to stimulate bone repair in other locations around the body.

The overall goal of this procedure is to successfully resect a portion of bone from the rib of a mouse. The procedure was developed as a model to study large-scale long bone repair.

外科手术切除为鼠标肋骨:模型进行大规模的长骨修复

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

腔内药物输送到鼠标动静脉瘘内皮

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

应用新型磁吻合技术 Suprahepatic 腔静脉重建大鼠原位肝移植模型的研究

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

中央静脉狭窄病的Murine模型,使用具有流出狭窄病的动脉静脉瘘管病

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

Murine Surgical Model of Topical Elastase Induced Descending Thoracic Aortic Aneurysm

According to the Center for Disease Control, aortic aneurysms (AAs) were considered a leading cause of death in all races and both sexes from 1999-2016. An aneurysm forms as a result of progressive weakening and eventual dilation of the aorta, which can rupture or tear once it reaches a critical diameter. Aneurysms of the descending aorta in the chest, called descending thoracic aortic aneurysms (dTAA), make up a large proportion of aneurysm cases in the United States. Uncontained dTAA rupture is almost universally lethal, and elective repair has a high rate of morbidity and mortality. The purpose of our model is to study dTAA specifically, to elucidate the pathophysiology of dTAA and to search for molecular targets to halt the growth or reduce the size of dTAA. By having a murine model to study thoracic pathology precisely, targeted therapies can be developed to specifically test dTAA. The method is based on the placement of porcine pancreatic elastase (PPE) directly on the outer murine aortic wall after surgical exposure. This creates a destructive and inflammatory reaction, which weakens the aortic wall and allows for aneurysm formation over weeks to months. Though murine models possess limitations, our dTAA model produces robust aneurysms of predictable size. Furthermore, this model can be used to test genetic and pharmaceutical targets which may arrest dTAA growth or prevent rupture. In human patients, interventions such as these could help avoid aneurysm rupture, and difficult surgical intervention.

We describe a surgical protocol to consistently induce robust descending thoracic aortic aneurysms in mice. The procedure involves left thoracotomy, thoracic aorta exposure, and placement of a sponge soaked in porcine pancreatic elastase on the aortic wall.

局部性乳酸酶诱导下降胸腔主动脉瘤的毛尿外科模型

According to the Center for Disease Control, aortic aneurysms (AAs) were considered a leading cause of death in all races and both sexes from 1999-2016. ...

Creating Radio-cephalic Arteriovenous Fistula in the Forearm with a Modified No-Touch Technique

Autologous arteriovenous fistula (AVF) is the primary and best option to obtain vascular access for hemodialysis treatment; other options are arteriovenous graft (AVG) and central venous catheterization (CVC). The implementation of radio-cephalic autologous arteriovenous fistula (RC-AVF) in the forearm was preferred among patients with superior vascular conditions. However, there is a high rate of early fistula failure. The chosen surgical method is understood to have an effect on the maturation of the fistula. New surgical procedures such as radial artery deviation and reimplantation (RADAR) have been significantly improved for juxta-anastomotic stenosis. Nevertheless, new problems such as stenosis of arteries and narrowing of surgical indication were also found. In this report, we presented a modified no-touch technique (MNTT) to create an RC-AVF, in which the venous and arterial wall avoid devascularization and the radial artery does not sever.

We present a modified no-touch technique (MNTT) to create a radio-cephalic arteriovenous fistula (RC-AVF) in which the venous and arterial wall avoid devascularization and the radial artery does not sever.