The work was supported in part by funding from Alberta Livestock and Meat Agency and Natural Science and Research Council Discovery grant to S. D. Proctor.

在视频稿件和描述都实验动物的所有程序被机构动物护理和使用委员会批准，其次是动物保健的加拿大理事会确定的指导方针。 1.手术麻醉及新生儿猪手术准备<ol><li>在一个分离的前厅，premedicate颈部与含有肌内镇静麻醉药鸡尾酒底部附近25千克猪:二甲苯胺噻嗪（0.3毫克/千克），盐酸氯胺酮（10毫克/千克），右美托咪（15微克/千克）。 注:在麻醉前药物鸡尾酒增强术中疼痛控制添加丁丙诺啡（0.005-0.02毫克/千克）。 </li><li>麻醉与吸入异氟烷吸入气体的猪（4-5％异氟醚在500毫升千毫升/分钟O 2）使用面罩。使用兽医喉范围（17-25厘米长的直叶片）可视化的声带，并应用外用10％的利多卡因绝育的声音线。允许利多卡因喷30-60秒前联系声带气管插管，以减少声带痉挛及气道阻塞的可能性。 </li><li>由声带之间传递铐气管插管（5.0-7.0毫米内径（ID））插管猪并保持与异氟醚气体麻醉（0.5-2.0％异氟醚在1000-2000毫升/分钟O 2）使用关闭电路在整个手术麻醉重复吸入系统。评估麻醉由颚音调水平，并且两个踏板和眼睑反射响应。过期麻醉气体被清理和手术套件之外排出。 </li><li>清洁用2.0％氯己定外科擦洗液耳后跟一个70％异丙醇冲洗的外表面。用20G的静脉导管，导尿耳静脉，以提供静脉注射液（乳酸林格氏液; 5-10毫升/千克/小时）的手术过程中。安全脉搏血氧仪与MED舌粘膜表面的iCal胶带监测心脏速率和外周血氧合的饱和度（ 血氧饱和度）。 </li><li>放置在一个背卧位麻醉猪和剃从中间胸部尾端腹侧腹部到耻骨的腹侧。有两个交替的2.0％洗必泰外科擦洗和消毒水洗清洁这个区域。 </li><li>转移猪麻醉到手术套件和应用70％异丙醇冲洗的最终手术擦洗，晾干，然后再披动物。 </li><li>插入直肠温度探头大约2-4厘米到直肠监测体温。放置在一个循环水加热垫猪保持在手术过程正常体温（38-40℃）。 </li><li>悬垂性与放置在腹部周围的上覆象限模式4毛巾窗帘猪。放置第一个悬垂横跨剑突，第二悬垂沿腹部约5的c外侧面米外侧腹部中线。放置第三悬垂横跨骨盆的回肠波峰和第四悬垂性，像第二悬垂（尽管在相反侧），沿着腹部约5cm外侧到腹部中线的外侧面放置。 </li><li>放置一个大表的悬垂性，具有狭缝开口允许进入手术部位，在底层的毛巾窗帘并覆盖猪和整个手术表。最终的褶皱是一次性免缝悬垂放置在大桌子悬垂性。 </li></ol> 2.腹部手术和导管肠淋巴干的<ol><li>使20cm的皮肤切口，用手术刀刀片以暴露下面的腹部肌肉。切割与单极电灼（20瓦设置）腹部肌肉层以暴露壁层腹膜。打开壁层腹膜的20cm的线段与梅岑鲍姆剪刀进入腹腔脏器和淋巴管。 </li><li&gt;将牵开器在手术切口的颅方面保持腹腔开放手术的持续时间。 </li><li>滋润所有组织用温水（37℃）的无菌盐水整个手术过程。轻轻抬起一大段小肠包括从腹腔结肠，盲肠，回肠和空肠和它exteriorize到猪的左翼访问上腹部，肝和淋巴管。在固定位置上的外置肠道额外的毛巾窗帘形成吊带轻轻支持肠。 </li><li>找到淋巴管，它奠定了约4厘米右肾静脉，腔静脉工头6厘米尾，中间-腹和胰腺22,36,37附近肝右叶内脏方面下的颅内侧。识别淋巴管作为并列门静脉36,37的右侧腹段的半透明结构。 </li><li>分离淋巴VESSEL从周围筋膜轻轻挑逗废除棉签涂抹附加的组织。一旦船只的横向方面从周围组织分离，创建容器下方的"隧道"口用细钝尖镊子。 </li><li>通过三项2-0丝线缝合用细镊子淋巴管下方。首先结扎最尾缝合闭塞，扩张和淋巴填充容器。目的地离开这个缝合相对长（4厘米）的端部到导管固定到淋巴管。放置另外两个缝线彼此隔开1.0厘米和大约1.0-1.5厘米颅到锁定尾缝合。离开这两个缝线具有"单一松散结扎领带"，以允许在导管的更快固定到容器。 注意:位于最尾端结扎缝线和中间缝合（两个颅缝的）之间的淋巴管的段为导尿站点。关于苏TURE材料，如果需要2-0聚乳糖缝线可替代2-0丝线缝合。 </li><li>切一小口与虹膜剪船和扩张细钝钳的容器。用斜切端插入该容器插入约1.0-1.5厘米专门的导管（4.06外径（OD）×2.31毫米ID），和扎两个颅缝以固定导管就位。使用尾缝线的长缝合线端部的导管固定到容器中。 </li><li>用大量温生理盐水冲洗外置肠，轻轻将其返回到腹腔，确保肠道正确的解剖定位。 </li><li> （从paralumbar窝腹侧5-10厘米）Exteriorize导管在左侧中间侧翼。使用手术刀切开皮肤，并从腹腔到皮肤表面传递套针来创建导管的外化的开口。使用大凯利钳从腹部CAV exteriorize导管穿过套管针开放性。 </li><li>用2-0聚乳糖缝合圆（锥形）针一个简单的连续缝合模式关闭壁层腹膜。用一个简单的间断缝合模式以2-0聚乳糖缝合关闭上圆针腹部肌肉层。 </li><li>用2-0缝合聚乳糖合皮肤的表皮下图案切割针。固定形象化导管与荷包缝合模式皮肤上切割针2-0尼龙缝合。 </li><li>将一个专门的夹克猪，同时仍麻醉以减轻其位置，并减少恢复期间猪的压力。 </li></ol> 3.手术后的恢复和淋巴收集<ol><li>吸入麻醉停产前约10分钟，辖bupenorphine（0.1毫克/千克）肌注提供即时的术后镇痛。继续bupenorphine（0.1毫克/千克），每12小时24〜48小时，以保持手术后镇痛。 </li><li>监控猪的手术后并发症7天期每8-12小时。 </li><li>收集在500毫升聚丙烯洗瓶淋巴液，涂有乙二胺四乙酸（EDTA）和补充有抗生素;青霉素（6000 IU），链霉素（6毫克）和两性霉素B（3毫克）为7天期间，每12小时。 </li></ol> 4.脂蛋白ApoB48，甘油三酯，胆固醇和总蛋白定量从淋巴收集<ol><li>离心淋巴样品在1800 XG在4℃下5分钟。收集上清液，并将其用于甘油三酯，胆固醇和总蛋白的定量。 </li><li>划分上清液分成三个样品:未稀释的样品，将样品以1:20稀释蒸馏水稀释1的最终样品:100用蒸馏水。 </li><li>使用未稀释的上清液来测量胆固醇水平，与市售的试剂盒。 </li><li>使用1:20和1:100稀释的样品以测量甘油三酯和总蛋白水平与市售的试剂盒和二辛可宁酸总蛋白分别测定。 </li></ol> 5.定量脂蛋白ApoB48，从淋巴收集38 <ol><li>确定脂蛋白ApoB48的浓度与适于免疫Western印迹方法38。用3-8％三 - 酯 - 十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分离总淋巴。 </li><li>转移分离的蛋白质，以聚偏二氟乙烯膜（0.45微米），用山羊多克隆抗体的ApoB（1:4000）孵育它们，然后将其绑定与抗山羊次级抗体。 </li><li>采用量化与啮齿类动物纯化蛋白质ApoB48标准的线性光密度比较脂蛋白ApoB48化学发光。 </li></ol>

The authors have nothing to disclose.

收集肠淋巴是调查参与脂质8-12和13-15的药物代谢，癌症转移16,17，贩卖细胞和免疫功能18-26机制，在各种实验动物模型的好方法。事实上，收获大量任一外围设备（传入）和中央（传出和大型行李箱血管）淋巴的较长时间内的能力一直是特别重要的理解发生在以下与免疫调节剂18-22挑战细胞群随时间的变化，25,26。同样，中央淋巴结收集已划定参与?...

淋巴系统是生理学的充分研究领域。淋巴导管的临床前模型发生在不同的动物物种1-8和由制药工业和研究机构用于研究参与脂质8-12和药物代谢13-15，与实验治疗17癌转移16的机制，及免疫功能18 -26。本研究旨在探讨在国内猪模型采用肠淋巴导管干线来衡量脂蛋白代谢的成分。脂蛋白代谢参与乳糜微粒的产生和分泌，以及在相关的脂质和总蛋白的变化。这些都是重要的考虑因素，因为有常用的啮齿动物模型与人类之间，因此脂质代谢的主要区别，采用猪模型，收集肠淋巴能为研究脂类我提供更多的可比信息代谢障碍，在人27-31。 若干外科技术用于收集在大动物种属的肠道淋巴:颅骨肩方法（ 即，胸导管导尿）5，横向上侧翼的方法32-34，和腹中线或正中的方法22,35。详细这段视频描述了使用的肠淋巴树干导管下腹正中手术方法在猪的外科手术。细心的手术技术许可淋巴导管的这种方法来收集大量淋巴结及其成员在一段时间过长。 该技术将打开的应用程序，许多学科研究的各种生理功能万千。应用可包括，但不限于，全身脂蛋白和脂质代谢，免疫监视，肿瘤发生和转移，肠道功能，并且developm耳鼻喉科和肠道炎症疾病的进展。

<table><tbody><tr><td>Miller laryngoscope blade</td><td>Welch Allyn</td><td>68044</td><td>182 mm length</td></tr><tr><td>Surgivet advisor: Vital signs monitor</td><td>Surgivet</td><td>V9203</td><td></td></tr><tr><td>Rectal temperature probe</td><td>Surgivet</td><td>V3417</td><td></td></tr><tr><td>Mono-polar electrosurgery generator</td><td>Valley Lab</td><td></td><td></td></tr><tr><td>Metzenbaum scissors</td><td>Fine Science</td><td>14518-18</td><td></td></tr><tr><td>Tuffier retractor</td><td>Stevens</td><td>162-11-676</td><td></td></tr><tr><td>Mosquito forceps</td><td>Stevens</td><td>162-7-10</td><td></td></tr><tr><td>Kelly forceps-curved (14cm)</td><td>Stevens</td><td>162-7-38</td><td></td></tr><tr><td>Allis tissue forceps</td><td>Stevens</td><td>162-7-38</td><td></td></tr><tr><td>Forceps dressing-eye (10.2cm)</td><td>Stevens</td><td>162-18-780</td><td></td></tr><tr><td>Forceps dressing-Adison (12.1cm)</td><td>Stevens</td><td>162-17-2510</td><td></td></tr><tr><td>Needle Drivers</td><td>Stevens</td><td>162-V98-42</td><td></td></tr><tr><td>Iris scissors</td><td>Fine science</td><td>14058-11</td><td></td></tr><tr><td>Circulating water pump</td><td>Jorvet</td><td>J-783X</td><td></td></tr><tr><td>Maxitherm-Vinyl blanket</td><td>Jorvet</td><td>J-784C</td><td></td></tr><tr><td>Q tip applicators</td><td>Fisher Scientific</td><td>22-037-960</td><td></td></tr><tr><td>Catheterization&amp;nbsp; tubing (4.06 OD X 2.31 ID)</td><td>Braintree Scientific Inc.</td><td>MRE-160</td><td>Micro-Renethane implantation tubing</td></tr><tr><td>2-0 silk suture</td><td>Ethicon</td><td>LA556</td><td></td></tr><tr><td>2-0 polyglactin suture</td><td>Ethicon</td><td>J443H</td><td>2-0 vicryl</td></tr><tr><td>Large animal jacket</td><td>Lomir Biomedical Inc.</td><td>SSJ2YC</td><td></td></tr><tr><td>Polypropylene wash bottles</td><td>Fisher Scientific</td><td>03-409-22C</td><td>500 ml</td></tr><tr><td>Penicillin-Streptomycin</td><td>Sigma Aldrich</td><td>D4333</td><td></td></tr><tr><td>EDTA</td><td>Sigma Aldrich</td><td>60-00-4</td><td></td></tr><tr><td>Amphotericin B</td><td>Sigma Aldrich</td><td>A2411</td><td></td></tr><tr><td>Azaperone</td><td>Elanco Animal Health</td><td></td><td>Stresnil</td></tr><tr><td>Dexmedetomidine hydrochloride</td><td>Zoetis</td><td>6295</td><td>Dexdomitor</td></tr><tr><td>Isoflurane</td><td>Abbott Animal&amp;nbsp; Health</td><td>05260-5</td><td>IsoFlo</td></tr><tr><td>Ketamine hydrochloride</td><td>Zoetis</td><td>2626</td><td>Ketaset</td></tr><tr><td>Bupenorphine hydrochloride</td><td>Champion Alstoe Animal Health</td><td></td><td>DIN:02347510</td></tr><tr><td>6 mm Endotracheal tube</td><td>Jorvet</td><td>J-165d</td><td></td></tr><tr><td>10% Lidocaine spray</td><td>AstraZeneca</td><td></td><td>DIN:02003767</td></tr><tr><td>4 % Chlorhexidine surgical scrub</td><td>Partnar Animal Health</td><td>PCH-011</td><td>Diluted: 2.0% solution</td></tr><tr><td>3M Surgical steri- drape</td><td>3M Health Care</td><td>1040</td><td></td></tr><tr><td>SDS page gel</td><td>Invitrogen</td><td>EA0375BOX</td><td>3-8 % tris acetate</td></tr><tr><td>Polyvinylidene fluoride membrane</td><td>Millipore</td><td>IPVH00010</td><td>0.45 &amp;mu;m pore size</td></tr><tr><td>ApoB antibody&amp;nbsp;</td><td>EMD Millipore</td><td>AB742</td><td>1:4000 dilution</td></tr><tr><td>Donkey anti-goat IgG-HRP</td><td>Santa Cruz Biotechnology</td><td>Sc-2304</td><td></td></tr><tr><td>ECL Prime Western Blotting Reagent</td><td>GE Healthcare LifeSciences</td><td>RPN2232&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td></tr><tr><td>Triglyceride Kit</td><td>Wako Pure Chemicals</td><td>998-40391/994-40491</td><td></td></tr><tr><td>Total Cholesterol Kit</td><td>Wako Pure Chemicals</td><td>439-17501</td><td></td></tr><tr><td>Total Protein</td><td>&amp;nbsp;Pierce&amp;nbsp;</td><td>23225</td><td>Bicinchoninic Acid Assay</td></tr></tbody></table>

long-term catheterization of the intestinal lymph trunk and collection of lymph in neonatal pigs

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal (central) efferent lymph. Importantly, the collection of central lymph from the intestine enables researchers to study both the mechanisms and lipid constitutes associated with lipid metabolism, intestinal inflammation and cancer metastasis, as well as cells involved in immune function and immunosurveillance. A ventral mid-line surgical approach permits excellent surgical exposure to the cranial abdomen and relatively easy access to the intestinal lymph trunk vessel that lies near the pancreas and the right ventral segment of the portal vein underneath the visceral aspect of the right liver lobe. The vessel is meticulously dissected and released from the surrounding fascia and then dilated with sutures allowing for insertion and subsequent securing of the catheter into the vessel. The catheter is exteriorized and approximately 1 L/24 hr of lymph is collected over a 7 day period. While this technique enables the collection of large quantities of central lymph over an extended period of time, the success depends on careful surgical dissection, tissue handling and close attention to proper surgical technique. This is particularly important with surgeries in young animals as the lymph vessels can easily tear, potentially leading to surgical and experimental failure. The video demonstrates an excellent surgical technique for the collection of intestinal lymph.

<html>仔猪肠淋巴干的淋巴导管允许大约1升的中央淋巴/ 24小时收集了7天的时间。在这个实验中收集的淋巴含有脂质代谢，即总淋巴蛋白，ApoB48脂蛋白，甘油三酯，总蛋白，和胆固醇的组分。表1列出代表数量从三头猪汇集淋巴结样品这些脂质成分的。值得注意的是，淋巴流和脂质成分是在符合由下列肠淋巴管的导管等研究者报告中央淋巴的值（淋巴流570±158 979±284毫升/ 24小...

Watch this Scientific Journal Video about Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs at JoVE.com

Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

在新生儿猪淋巴的肠淋巴躯干和收集的长期导尿

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal ...

long-term-catheterization-intestinal-lymph-trunk-collection-lymph

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

9.3K 次观看.  University of Alberta. We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

视频: Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

Measurement of Cytosolic Ca2+ in Isolated Contractile Lymphatics

Lymphatic vessels comprise a multifunctional transport system that maintains fluid homeostasis, delivers lipids to the central circulation, and acts as a surveillance system for potentially harmful antigens, optimizing mucosal immunity and adaptive immune responses1. Lymph is formed from interstitial fluid that enters blind-ended initial lymphatics, and then is transported against a pressure gradient in larger collecting lymphatics. Each collecting lymphatic is made up of a series of segments called lymphangions, separated by bicuspid valves that prevent backflow. Each lymphangion possesses a contractile cycle that propels lymph against a pressure gradient toward the central circulation2. This phasic contractile pattern is analogous to the cardiac cycle, with systolic and diastolic phases, and with a lower contraction frequency4. In addition, lymphatic smooth muscle generates tone and displays myogenic constriction and dilation in response to increases and decreases in luminal pressure, respectively5. A hybrid of molecular mechanisms that support both the phasic and tonic contractility of lymphatics are thus proposed.
Contraction of smooth muscle is generally regulated by the cytosolic Ca2+ concentration ([Ca2+]i) plus sensitivity to Ca2+, of the contractile elements in response to changes in the environment surrounding the cell6. [Ca2+]i is determined by the combination of the movement of Ca2+ through plasma membrane ligand or voltage gated Ca2+ channels and the release and uptake of Ca2+ from internal stores. Cytosolic Ca2+ binds to calmodulin and activates enzymes such as myosin light chain (MLC) kinase (MLCK), which in turn phosphorylates MLC leading to actin-myosin-mediated contraction8. However, the sensitivity of this pathway to Ca2+ can be regulated by the MLC phosphatase (MLCP)9. MLCP activity is regulated by Rho kinase (ROCK) and the myosin phosphatase inhibitor protein CPI-17.
Here, we present a method to evaluate changes in [Ca2+]i over time in isolated, perfused lymphatics in order to study Ca2+-dependent and Ca2+-sensitizing mechanisms of lymphatic smooth muscle contraction. Using isolated rat mesenteric collecting lymphatics we studied stretch-induced changes in [Ca2+]i and contractile activity. The isolated lymphatic model offers the advantage that pressure, flow, and the chemical composition of the bath solution can be tightly controlled. [Ca2+]i was determined by loading lymphatics with the ratiometric, Ca2+-binding dye Fura-2. These studies will provide a new approach to the broader problem of studying the different molecular mechanisms that regulate phasic contractions versus tonic constriction in lymphatic smooth muscle.

Measurement of Cytosolic Ca2+ in Isolated Contractile Lymphatics

We introduce an approach to evaluate the cytosolic Ca2+ concentration in isolated lymphatics to study Ca2+-dependent and Ca2+-sensitizing mechanisms of lymphatic smooth muscle contraction.

胞内钙的测量 2 +</sup在隔离的收缩淋巴管

Lymphatic vessels comprise a multifunctional transport system that maintains fluid homeostasis, delivers lipids to the central circulation, and acts as a ...

科研

免疫与感染

Catheterization of Intestinal Loops in Ruminants

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the outside world. As such, the intestine plays a pivotal role in immunosurveillance and protection from enteric pathogens. Investigating intestinal physiology and immunology commonly employs 'intestinal loops' as an experimental model. The majority of these loop models are non-recovery surgical procedures that study short-term (&lt;24 hr) changes in the intestine (1-3). We previously created a recovery intestinal loop model to specifically measure long-term (&lt;6 mo) immunological changes in the intestine of sheep following exposure to vaccines, adjuvants, and viruses (4). This procedure localized treatments to a specific 'loop', allowing us to sample this area of the intestine. A significant drawback of this method is the single window of opportunity to administer treatments (i.e. at the time of surgery). Furthermore, samples of both the intestinal mucosa and luminal contents can only be taken at the termination of the project. Other salient limitations of the above model are that the surgical manipulation and requisite post-operative measures (e.g. administration of antibiotics and analgesics) can directly affect the treatment itself and/or alter immune function, thereby confounding results. Therefore, we modified our intestinal loop model by inserting long-term catheters into the loops. Sheep recover fully from the procedure, and are unaffected by the exteriorized catheters. Notably, the establishment of catheters in loops allows us to introduce multiple treatments over an extended interval, following recovery from surgery and clearance of drugs administered during surgery and the post-operative period.

We describe a novel surgical method for catheterizing 'intestinal loops' within the ileum of sheep. Once animals have recovered from surgery and have cleared antibiotics and analgesics, multiple treatments can be deposited directly in loops via the catheters.

导尿在反刍动物的肠袢

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the ...

生物学

The Mesenteric Lymph Duct Cannulated Rat Model: Application to the Assessment of Intestinal Lymphatic Drug Transport

The intestinal lymphatic system plays key roles in fluid transport, lipid absorption and immune function. Lymph flows directly from the small intestine via a series of lymphatic vessels and nodes that converge at the superior mesenteric lymph duct. Cannulation of the mesenteric lymph duct thus enables the collection of mesenteric lymph flowing from the intestine. Mesenteric lymph consists of a cellular fraction of immune cells (99% lymphocytes), aqueous fraction (fluid, peptides and proteins such as cytokines and gut hormones) and lipoprotein fraction (lipids, lipophilic molecules and apo-proteins). The mesenteric lymph duct cannulation model can therefore be used to measure the concentration and rate of transport of a range of factors from the intestine via the lymphatic system. Changes to these factors in response to different challenges (e.g., diets, antigens, drugs) and in disease (e.g., inflammatory bowel disease, HIV, diabetes) can also be determined. An area of expanding interest is the role of lymphatic transport in the absorption of orally administered lipophilic drugs and prodrugs that associate with intestinal lipid absorption pathways. Here we describe, in detail, a mesenteric lymph duct cannulated rat model which enables evaluation of the rate and extent of lipid and drug transport via the lymphatic system for several hours following intestinal delivery. The method is easily adaptable to the measurement of other parameters in lymph. We provide detailed descriptions of the difficulties that may be encountered when establishing this complex surgical method, as well as representative data from failed and successful experiments to provide instruction on how to confirm experimental success and interpret the data obtained.

Here we describe a technique to cannulate the mesenteric lymph duct in rats which enables quantification of lipid and drug transport via the lymphatic system following intestinal delivery. The technique can be adapted to assess mesenteric lymph concentrations and/or transport of fluid, immune cells, peptides, proteins and lipophilic molecules.

在肠系膜淋巴管空心​​大鼠模型:应用肠淋巴药物运输的评估

The intestinal lymphatic system plays key roles in fluid transport, lipid absorption and immune function. Lymph flows directly from the small intestine ...

Microscopic Observation of Lymphocyte Dynamics in Rat Peyer's Patches

Na&#239;ve lymphocytes recirculate from the blood to the lymphoid tissues under physiological condition and it is commonly recognized as an important phenomenon in the gut immunity. The stroma of secondary lymphoid organs, such as Peyer&#8217;s patches (PPs) or mesenteric lymph nodes, are where na&#239;ve lymphocytes sense antigens. Na&#239;ve lymphocytes circulate through the bloodstream to reach high endothelial venules, the portal of entry into PPs. Some immunomodulators are estimated to influence lymphocyte migration, but the precise evaluation of microcirculation dynamics is very difficult, and establishing a method to observe lymphocyte migration in vivo can contribute to the clarification of the precise mechanisms. We refined the method of collecting lymphocytes from the lymph duct and observing the detailed dynamics of gut-tropic lymphocytes in rat PPs. We chose confocal laser scanning microscopy to observe rat PPs in vivo and recorded it using time-lapse photography. We can now obtain clear images that can contribute to the analysis of lymphocyte dynamics.

Here, we describe a precise method for collecting thoracic duct lymphocytes and observing the migration of gut-tropic lymphocytes in rat Peyer&#8217;s patches for 3 hours using time-lapse photography. This technique can clarify how the dynamics of lymphocytes are affected under inflammatory conditions.

鼠佩耶斑块中淋巴细胞动力学的微观观察

Na&#239;ve lymphocytes recirculate from the blood to the lymphoid tissues under physiological condition and it is commonly recognized as an important ...

医学

Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries does not solely rely on the research team's surgical skills; perioperative care and management procedures, like intubation, central venous line, and arterial line placement, are necessary and of the utmost importance for favorable experiment results. As it is uncommon for research teams to have anesthesiologists or any other staff other than the surgical team, the surgical team involved in translational research must acquire and/or develop the skills to perform the perioperative care. The purpose of this paper is to show the techniques of intubation, central venous catheter, and arterial line placement used and perfected at the Toronto Organ Preservation Laboratory over the last 10 years, to be used as a reference for future researchers joining either this team or any other lab performing translational research protocols in swine and/or abdominal transplantation.

<meta charset="utf8"></head><body>猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

To obtain the best possible results, surgeons performing translational research experiments should be proficient at intubation and vascular line placement. This paper describes the techniques used by the Toronto Organ Preservation Laboratory to perform these procedures.

猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries ...

用于猪长期血管通路的 Hickman 导管

Hickman Catheter Use for Long-Term Vascular Access in a Preclinical Swine Model

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood monitoring and reliable intravenous fluid and drug administration. Specifically, the tunneled multi-lumen Hickman catheter (HC) is commonly used in swine models due to its lower extrication and complication rates. Despite fewer complications relative to other CVCs, HC-related morbidity presents a significant challenge, as it can significantly delay or otherwise negatively impact ongoing studies. The proper insertion and maintenance of HCs is paramount in preventing these complications, but there is no consensus on best practices. The purpose of this protocol is to comprehensively describe an approach for the insertion and maintenance of a tunneled HC in swine that mitigates HC-related complications and morbidity. The use of these techniques in &gt;100 swine has resulted in complication-free patent lines up to 8 months and no catheter-related mortality or infection of the ventral surgical site. This protocol offers a method to optimize the lifespan of the HC and guidance for approaching issues during use.

作者聚焦：Hickman 导管在临床前猪模型中用于长期血管通路  

A reliable and reproducible approach for the insertion and maintenance of a tunneled Hickman catheter for long-term vascular access in swine is described. Placement of a central venous catheter allows for convenient daily sampling of whole blood from awake animals and intravenous administration of medication and fluids.

Hickman 导管在临床前猪模型中用于长期血管通路

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood ...

Surgical Technique for Lumbar Spinal Catheter Insertion in Pigs Enabling Continuous Access to the Thecal Sac in a Terminal Setup

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine and human central nervous systems (CNS). However, accessing the cerebrospinal fluid (CSF) in larger pig breeds by conventional lumbar puncture techniques can be challenging due to an oblique orientation of the spinal spinous processes and a limited interlaminar space. Accordingly, an open surgical procedure for inserting a lumbar spinal catheter for continuous CSF sampling at the L4/L5 level in pigs is thoroughly described in this work. After positioning the pig and identifying the anatomical landmarks, a dorsal midline surgical incision is made to expose the spinous processes. By advancing the introducer needle, the spinal catheter is inserted inside the thecal sac of the spinal canal while leaving the bone structures of the spine intact. This method allows continuous infusion into or sampling from the porcine thecal sac with minimal bleeding or CSF leakage. The procedure is simple, time-efficient, and reproducible across different experimental setups, offering significant potential for various pre-clinical studies, including pharmacokinetic research, surgical training, and spinal cord injury models.

We present a technique for inserting a lumbar spinal catheter at the L4-L5 level in a 3-month-old Danish Landrace pig as part of a terminal research protocol, enabling continuous infusion or CSF sampling from the thecal sac.

猪腰椎导管插入手术技术，可在终端设置中连续进入鞘囊

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine ...

在新生儿猪淋巴的肠淋巴躯干和收集的长期导尿

摘要

探索更多视频

胞内钙的测量^{2 +}

导尿在反刍动物的肠袢

在肠系膜淋巴管空心大鼠模型:应用肠淋巴药物运输的评估

鼠佩耶斑块中淋巴细胞动力学的微观观察

猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

Hickman 导管在临床前猪模型中用于长期血管通路

Hickman 导管在临床前猪模型中用于长期血管通路

Hickman 导管在临床前猪模型中用于长期血管通路

猪腰椎导管插入手术技术，可在终端设置中连续进入鞘囊

在新生儿猪淋巴的肠淋巴躯干和收集的长期导尿

摘要

探索更多视频

胞内钙的测量 2 +

导尿在反刍动物的肠袢

在肠系膜淋巴管空心​​大鼠模型:应用肠淋巴药物运输的评估

鼠佩耶斑块中淋巴细胞动力学的微观观察

猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

猪插管、中心静脉导管和动脉导管置入用于腹部移植手术的转化研究

Hickman 导管在临床前猪模型中用于长期血管通路

Hickman 导管在临床前猪模型中用于长期血管通路

Hickman 导管在临床前猪模型中用于长期血管通路

猪腰椎导管插入手术技术，可在终端设置中连续进入鞘囊

胞内钙的测量^{2 +}

在肠系膜淋巴管空心大鼠模型:应用肠淋巴药物运输的评估