Name: murine ileocolic bowel resection with primary anastomosis
Uploaded: 2014-10-29T14:00:00.000+00:00
Duration: 8 min 49 s
Description: Watch this Scientific Journal Video about Murine Ileocolic Bowel Resection with Primary Anastomosis at JoVE.com

We would like to acknowledge the funding contributions of the Canadian Surgical Research Fund, the Edmonton Civic Employees Research Assistance Fund, and the Alberta IBD Consortium through a grant from Alberta Innovates.

Tiernutzung Protokolle wurden von der Health Science Animal Care und Verwenden Ausschuss an der Universität von Alberta genehmigt. 1. Herstellung der Instrumente, Tiere und Operative-Setup <ol><li> Übertragen Tieren zu einem neuen, sauberen Käfig abwesend aller feste Nahrung 24 Stunden vor dem Eingriff. Sie können den freien Zugang zu Wasser und flüssige Diät ad lib bis zum Zeitpunkt des Verfahrens. </li><li> Autoklav alle für das Verfahren erforderlichen Instrumente. Saubere Arbeitsfläche und Narkosenasenkegel mit 70% Ethanol. </li><li> Richten Sie die Arbeitsfläche mit Operationsmikroskop, Narkosegerät und Zubehör in einer Weise, die angenehm für den Operateur ist. Die Ellenbogen des Chirurgen sollte erlaubt werden, um bequem auf dem OP-Tisch liegen, mit Händen und Armen unbehindert durch Ausrüstung. Instrumente, Nahtmaterial, Wattestäbchen und eine 10-ml-Spritze sollte an einem Ort, die einen einfachen Zugang während des Verfahrens ermöglicht platziert werden. </li><li> EinrichtenOverhead-Wärmelampen, sowohl Wärme für das Tier während des Verfahrens und Licht für die Arbeitsfläche bereitzustellen. </li><li> Füllen Sie eine 50 ml konischen Röhrchen mit 0,9% Kochsalzlösung und eine 1,5-ml-Röhrchen mit Vaseline und Ort in der Nähe der Arbeitsfläche. HINWEIS: Sterilisieren alle Instrumente und OP-Bedarf. Da der Darm durchtrennt, ist das Verfahren selbst nicht steril. Es wird als Rein verunreinigt. Ergreifen Sie Maßnahmen, um die Einführung von exogenen Quellen von Infektionen zu vermeiden. </li></ol> 2. Ileocolische Resektion mit Anastomose <ol><li> Anästhesie zu induzieren durch Verabreichen von 4% Isofluran mit einer Sauerstofffließgeschwindigkeit von 2 L / min über Nasenkegel von der Isofluran-Verdampfer, bis die Tiere Atmungsrate verlangsamt etwa 30-40 Atemzüge / min. Mit mäßigem Druck auf den Hinterfuß der Maus, um sicherzustellen, gibt es keine Schmerzreaktion vor der Einleitung des Verfahrens. An dieser Stelle, drehen Sie Isofluran bis 2% und Sauerstofffluss auf 0,5 l / min. IntermittierendCheck Schmerzreaktion während des Verfahrens und stellen Isofluran Flussrate entsprechend. </li><li> Vaseline in die Augen, um das Trocknen während der Operation zu verhindern und zu immobilisieren die Maus in die Rückenlage mit Gliedmaßen mit durchsichtigem Klebeband gesichert. </li><li> Reinigen Sie den Bauch mit povodine / Jodlösung, und wechseln Sie in neue sterile Handschuhe. </li><li> Machen Sie eine 1,5 cm Hautschnitt in der oberen Mittellinie des Bauches mit scharfen Punkt Dissektionsscheren die Faszien und Peritoneum aussetzen. Öffnen Sie die Faszien / Bauchfellschicht in ähnlicher Weise durch die Linea alba in die Bauch Inhalten aussetzen. </li><li> Im Gegensatz zum Menschen, wird die Maus Blinddarm typischerweise im linken oberen Quadranten des Abdomens. Einmal identifiziert, sanft erfassen die Blinddarm mit einer Pinzette und bringt sie durch den Schnitt. Verwenden feuchten Wattetupfer auffächern etwa 3 cm von terminalen Ileum, der sich vom Blinddarm über eine sterile Gaze auf der Bauchfläche (1A) drapiert.Sicherzustellen, dass die freiliegenden Darm während der Gesamtheit des Verfahrens feucht mit 0,9% Kochsalzlösung aufbewahrt. </li><li> Identifizieren Sie die ileozökalen Arterie abzweigt A. mesenterica superior entlang des Dickdarms (1A) mit dem Operationsmikroskop. Präparieren Sie sich die avaskuläre Gewebe neben dem ileozökalen Arterie, einzukreisen und zu ligieren die Arterie mit einem 5-0 Seidenkrawatte. Nächstes den regionalen Blutversorgung des terminalen Ileum und wählen Sie eine Durchtrennung Punkt 1,5-2 cm proximal der ileozökalen Kreuzung. Ligieren die Zweige zu diesem Abschnitt des Ileum wie oben. Teilen Sie die Arterien mit Mikro Dissektionsscheren. </li><li> Teilen Sie die ischämische Bereiche des Ileum und Colon gewährleisten eine ausreichende Blutzufuhr zu den durchtrennten Enden (Abbildung 1B). Es ist oft hilfreich, um die Ileum durch Teilung in einem 30 Grad Winkel, um den Durchmesser des Lumens zu erhöhen, damit es den Dickdarm eher entspricht spatelförmig. Sobald der ileozökalen Teil Darm entfernt wurde, ausrichtentrennten Enden des Ileum und Colon auf die Gaze, die Gewährleistung der mesenterialen Grenzen jeweils ausgerichtet sind. </li><li> Konstruieren Sie die Anastomose durch Annähern der durchtrennten Ende Ileum zum durchtrennten Ende Dick Verwendung unterbrochen 8-0 Polypropylen Nähte (Abbildung 1c). Der erste Stich wird an der mesenterialen Rand platziert, mit anschließender Nähte platziert alle 0,5 mm, bis die Ileocolische Anastomose wasserdicht ist. Beim Passieren der Nähnadel durch das Ileum und Kolon, sicherzustellen, dass die Schnittkante nicht gerollt wird, und die Nadelstiche sind 0,5 mm von den Schnittkanten des Darms. Eine typische Anastomose erfordert 14 bis 16 Einzelnähten. Testen Sie die Integrität und die Durchgängigkeit der Anastomose nach Fertigstellung durch Walzen ein Wattestäbchen proximal nach distal über das Ileum, Inhalte durch die Anastomose zu erzwingen. Dünndarminhalt sollte sich frei in den Dickdarm, ohne Anastomoseninsuffizienz geben. </li><li> Spülung des belichteten Darm mit 3-4 ml 0,9% iger Kochsalzlösung aus der 10 ml Spritzevon der Oberfläche des Darms abzuwaschen Stuhl, und liefern Darm wieder in die Bauchhöhle. Mit 2 ml 0,9% Kochsalzlösung spülen Bauchhöhle, und dann abtropfen lassen diese Flüssigkeit durch leichten Druck auf die Bauchwand seitlich. </li><li> Schließen Sie den Schnitt mit einem 3-0 Seidenfortlaufende Naht, und die Strömung von Isofluran einzustellen. Verwalten 0,1 mg / kg des lang wirkenden Opiat -buprenorphine- subkutan zur postoperativen Schmerzkontrolle. </li><li> Beachten Sie die Tiere unter der Wärmelampe, bis sie sind mobil übertragen sie dann auf eine kontinuierlich erwärmt Käfig. </li></ol> 3. Postoperative Pflege und Überwachung <ol><li> Überwachung Tiere in einem kontinuierlich erwärmt Käfig für Anzeichen von Leiden für den Rest des Tages. Übertragen Tiere zurück in die Tierstation in einer neuen sterilen Käfig mit Zugang zu flüssigen Nahrung und Wasser ad lib. Die Tiere können in Gruppen von 3-4 untergebracht werden. </li><li> Führen Sie eine Überprüfung der postoperativen Tiere am nächsten Morgen,sicherzustellen, dass die Tiere nicht in der Not erscheinen. Feed-nur flüssige Nahrung. Wenn sie unbequem (dh gekrümmte Haltung oder minimale Aktivität) erscheinen zu verwalten eine zusätzliche Dosis von subkutan Buprenorphin. Überprüfen Sie auf die Tiere noch einmal am Nachmittag am postoperativen Tag 1. </li><li> Am Morgen des postoperativen Tag 2 sollte die Tiere vollständig erholt erscheinen. Der Nachweis der Lebensmittelkonsum und stooling sind positive Anzeichen für eine Erholung. Nun, wieder eine solide Chow-Diät für die Tiere. HINWEIS: Anzeichen von Leiden sind gekrümmte Haltung, schlechte Pflege und minimale Aktivität. Wenn Anzeichen von Leiden sind prominente sollten die Tiere eingeschläfert werden. </li><li> Euthanize die Tiere durch Induktion tiefe Narkose mit 4% Isofluran in einem O 2 Strömungsgeschwindigkeit von 2 L / min, bis Tiere reagieren nicht auf Fußdruck. Führen Genickbruch, und beobachten Sie auf Anzeichen von richtigen Euthanasie. </li><li> Richtlinien variieren, so finden Sie in den Institutionen Empfehlungen für Indikationenfür und geeignete Methoden der Euthanasie bei Mäusen. </li></ol>

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American journal of physiology.</a> Gastrointestinal and liver physiology. 293 (5), (2007).</li><li>Speck, K. E., De Cruz, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation+enhances+resection-induced+intestinal+adaptive+growth+in+IL-10+null+mice.">Inflammation enhances resection-induced intestinal adaptive growth in IL-10 null mice.</a> The Journal of surgical research. 168 (1), 62-69 (2011).</li><li>Rigby, R. J., Hunt, M. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+animal+model+of+postsurgical+bowel+inflammation+and+fibrosis:+the+effect+of+commensal+microflora.">A new animal model of postsurgical bowel inflammation and fibrosis: the effect of commensal microflora.</a> Gut. 58 (8), 1104-1112 (2009).</li><li>Kostic, A. D., Xavier, R. J., Gevers, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Microbiome+in+Inflammatory+Bowel+Disease:+Current+Status+and+the+Future+Ahead.">The Microbiome in Inflammatory Bowel Disease: Current Status and the Future Ahead.</a> Gastroenterology. , 1-11 (2014).</li><li>Gevers, D., Kugathasan, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Treatment-Naive+Microbiome+in+New-Onset+Crohn&#39;s+Disease.">The Treatment-Naive Microbiome in New-Onset Crohn&#39;s Disease.</a> Cell host &amp; microbe. 15 (3), 382-392 (2014).</li><li>Ahmed, T., Rieder, F., Fiocchi, C., Achkar, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+postoperative+recurrence+in+Crohn&#39;s+disease.">Pathogenesis of postoperative recurrence in Crohn&#39;s disease.</a> Gut. 60 (4), 553-562 (2011).</li><li>Borowiec, A., Sydora, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+bowel+fibrosis+and+systemic+inflammatory+response+after+ileocolonic+anastomosis+in+IL-10+null+mice.">Small bowel fibrosis and systemic inflammatory response after ileocolonic anastomosis in IL-10 null mice.</a> Journal of Surgical Research. 178 (1), 147-154 (2012).</li><li>Devine, A. A., Gonzalez, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Impact of Ileocecal Resection and Concomitant Antibiotics on the Microbiome of the Murine Jejunum and Colon. 8 (8), (2013).</li><li>Bij, G. J., Oosterling, S. J., Beelen, R. H. J., Meijer, S., Coffey, J. C., van Egmond, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perioperative+period+is+an+underutilized+window+of+therapeutic+opportunity+in+patients+with+colorectal+cancer.">The perioperative period is an underutilized window of therapeutic opportunity in patients with colorectal cancer.</a> Annals of surgery. 249 (5), (2009).</li><li>Scott, A. D., Uff, C., Phillips, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+macrophage+function+by+suture+materials+and+anastomotic+recurrence+of+Crohn&#39;s+disease.">Suppression of macrophage function by suture materials and anastomotic recurrence of Crohn&#39;s disease.</a> The British journal of surgery. 80 (3), (1993).</li><li>Kiernan, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+anastomosis+in+the+rat+facilitated+by+a+rapidly+digested+internal+splint+and+indigestible+but+absorbable+sutures.">Intestinal anastomosis in the rat facilitated by a rapidly digested internal splint and indigestible but absorbable sutures.</a> Journal of Surgical Research. 45, 427-431 (1988).</li><li>Andersen, T. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Action+of+matrix+metalloproteinases+at+restricted+sites+in+colon+anastomosis+repair:+an+immunohistochemical+and+biochemical+study.">Action of matrix metalloproteinases at restricted sites in colon anastomosis repair: an immunohistochemical and biochemical study.</a> Surgery. 140 (1), 72-82 (2006).</li><li>Thompson, S. K., Chang, E. Y., Jobe, B. 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+review:+Healing+in+gastrointestinal+anastomoses,+Part+I.">Clinical review: Healing in gastrointestinal anastomoses, Part I.</a> Microsurgery. 26 (3), 131-136 (2006).</li></ol>

The authors have nothing to disclose

Die murine ICR ist ein leistungsfähiges Modell, das verwendet werden kann, um die Auswirkungen der Operation bei Darmerkrankungen zu untersuchen. Dieser Artikel beschreibt ein Verfahren zur Durchführung ICR bei Mäusen mit einer Erfolgsquote von 95% und keine Probleme mit Gedeihstörungen wie durch stabile Gewichte reflektiert bis zu 28 Tage nach dem Eingriff. Die wichtigsten Herausforderungen für die erfolgreiche ICR gehören die Vermeidung Darm Hindernisse an der Anastomose und Anastomosen Lecks. <p class="jove...

Ileocolic resection (ICR) is a common procedure performed in both emergent and elective situations for a variety of illnesses. Crohn&#8217;s disease and colon cancers are the two most common indications for ICR. In both illnesses, recurrence in the bowel at the site of surgery represents a major clinical problem. Local recurrence rates for colon cancer remain an issue even with the most aggressive resections1. Following ICR in Crohn&#8217;s disease, the disease most frequently (in up to 80%) recurs in the neo-terminal ileum at 1 year after surgery2. Given the impact of these two illnesses and their recurrence after surgery, it is important to understand local intestinal factors after ICR, which may have intrinsic influences on the natural history of these diseases. Further, it is also important to consider anastomotic healing after ICR. In the early post-operative period, anastomotic leaks can have devastating consequences for patients resulting in repeat surgeries, stoma creations, significant morbidity, and even mortality3. Despite the importance of this topic, our current understanding of anastomotic healing remains in its infancy as a subject of research. Animal models of ICR, in particular mice, are an excellent platform for studying the intestinal and anastomotic healing following surgery.
A mouse model of ICR was initially developed by Helmrath et al. to be used as a model of short gut syndrome4. The authors experimented with various diet regiments and suture sizes to optimize animal survival following ICR. They concluded that feeding with liquid diet in the perioperative period and using 9-0 monofilament sutures resulted in an optimal post-operative survival of 88%. Since this initial publication, ICR in mice removing 50% of the small bowel has been used in several studies to explore the dynamics of massive small bowel resection and the adaptive growth response in attempt to develop new therapies for short gut syndrome5,6.
The first application of the ICR mouse model to Crohn&#8217;s disease used the IL-10-/- mouse model, which spontaneously develops colitis7. The authors found that after ICR these animals developed inflammation in the neo-terminal ileum similar to that seen in post-operative Crohn&#8217;s disease patients, and that this inflammation was dependent on the presence of bacteria7. More recently, this model was used to explore bacterial changes induced by ICR. In Crohn&#8217;s disease there is an associated dysbiosis with relative decreases in bacteria known to have anti-inflammatory properties and increases in invasive species of bacteria8,9. The association holds true in cases of post-operative recurrence10. Two studies sought to identify microbial changes resulting from ICR. The first used IL-10 null mice, and performed denaturing gel electrophoresis to compare bacterial similarity between the small bowel and colon after ICR11. This study demonstrated that bacterial populations became similar in the small intestine and colon following ICR. A subsequent study used wild type mice and 16s pyrosequencing for phylogenetic classification of bacterial species post-operatively. This study demonstrated a marked shift in bacterial species resulting from surgery alone with Clostridium species becoming dominant as well as an increase in &#978;-proteobacteria. The results also confirmed the findings of the previous study with similar populations found in the small bowel and colon after ICR12. 
ICR is a common procedure for patients with colon cancer involving the cecum and ascending colon, and it is becoming increasingly recognized that the host-response to surgery likely contributes to both local and distant tumor recurrence13. Despite this observation, models of ICR have not been utilized for the study of colorectal cancer and post-operative recurrence. Understanding both the systemic and local immunologic changes resulting from ICR will be important in investigating future therapies. Potential pathways involved in cancer recurrence post ICR include up regulation of growth factors, which may rescue cells from apoptosis and stimulate proliferation, mechanical tumor disruption with cell shedding, and loss of immune surveillance through post-operative immunosuppression13,14. 
Mouse models of ICR have the potential to be a powerful tool for the investigation of short bowel syndrome, Crohn&#8217;s disease, and colon cancer. They may also provide lessons on how to prevent early post-operative anastomotic complications by further defining the cellular and biochemical pathways involved in healing the newly constructed anastomosis. A major barrier in utilizing the murine ICR model is the technical difficulty. The intestinal anastomosis requires the use of 8-0 or 9-0 suture, an operating microscope, and training in microsurgical techniques. The goal of this article is to provide clear instructions on how to perform ICR in mice with the goal of utilizing this procedure in models of disease.

<table><tbody><tr><td>LD101 liquid rodent diet</td><td>testdiet.com</td><td></td><td></td></tr><tr><td>0.9% NaCl</td><td>Baxter</td><td>FKE1324</td><td>Injection quality saline</td></tr><tr><td>Operating Microscope</td><td>Ziess</td><td></td><td></td></tr><tr><td>Isoflurane Anesthetic Vaporizer</td><td>Harvard Apparatus</td><td>34-0483</td><td></td></tr><tr><td>Isoflurane</td><td>Abbott&amp;nbsp;</td><td>05260-05</td><td></td></tr><tr><td>Glass plate</td><td></td><td></td><td>For operating surface</td></tr><tr><td>Cotton swabs</td><td></td><td></td><td></td></tr><tr><td>Micro Castroviejo Needle holder, curved</td><td>World Precision Instruments</td><td>503377</td><td></td></tr><tr><td>Castroviejo straight scissors</td><td>World Precision Instruments</td><td>555530S</td><td></td></tr><tr><td>Dissecting Scissors</td><td>World Precision Instruments</td><td>15922</td><td></td></tr><tr><td>Dressing Forceps x 2</td><td>World Precision Instruments</td><td>500363</td><td></td></tr><tr><td>5-0 silk pre-cut sutures</td><td>Ethicon</td><td>A182H</td><td>For vessel ligation</td></tr><tr><td>8-0 Prolene on BV130-5 needle</td><td>Ethicon</td><td>8732H</td><td>For anastomosis</td></tr><tr><td>3-0 Silk on FS-2 needls</td><td>Ethicon</td><td>8665G</td><td>For abdominal wall closure</td></tr><tr><td>Petroleum Jelly</td><td>Vaseline</td><td></td><td></td></tr><tr><td>10 ml syringe</td><td>BD biosciences</td><td></td><td></td></tr><tr><td>Povidone-iodine 7.5% surgical Scrub</td><td>&lt;a href=&quot;http://betadine.com&quot;&gt;betadine.com&lt;/a&gt;</td><td></td><td></td></tr><tr><td>Heat lamps</td><td></td><td></td><td></td></tr><tr><td>buprenorphine 0.3 mg/ml</td><td>Reckitt Benckiser Healthcare Ltd.&amp;nbsp;</td><td>PL36699/0006</td><td></td></tr></tbody></table>

murine ileocolic bowel resection with primary anastomosis

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer being two common examples. Despite the frequency of these procedures, a significant knowledge gap remains in describing the inherent effects of intestinal resection on host physiology and disease pathophysiology. This article provides detailed instructions for an ileocolic resection with primary end-to-end anastomosis in mice, as well as essential aspects of peri-operative care to maximize post-operative success. When followed closely, this procedure yields a 95% long-term survival rate, no failure to thrive, and minimizes post-operative complications of bowel obstruction and anastomotic leak. The technical challenges of performing the procedure in mice are a barrier to its wide spread use in research. The skills described in this article can be acquired without previous surgical experience. Once mastered, the murine ileocolic resection procedure will provide a reproducible tool for studying the effects of intestinal resection in models of human disease.

Sterblichkeit und Gewichtsveränderung nach der OP. Die Sterblichkeitsraten folgenden ICR in 129S1 Wildtyp-Mäusen in der Regel ~ 5%. Die häufigste Ursache der Moral ist Darmverschluss an der Anastomose. Andere Todesursachen gehören Anastomoseninsuffizienz und interne Hernie, die zu Verstopfung Darm. Gewichtsverlust kann bis zu 14 Tage nach der Operation zu sehen ist, ist jedoch im allgemeinen nicht signifikant. Mäuse sind in der Regel präoperativen Gewicht komplett wiede...

Watch this Scientific Journal Video about Murine Ileocolic Bowel Resection with Primary Anastomosis at JoVE.com

Murine Ileocolic Bowel Resection with Primary Anastomosis

Ileocolic resection is commonly performed in several human diseases; however, little is known regarding the impact of intestinal resection on surgical illnesses. This article provides instruction on executing the procedure in mice with high success, providing a means to study the effects of ileocolic resection in models of disease.

Murine Ileocolische Darmresektion mit primärer Anastomose

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer ...

murine-ileocolic-bowel-resection-with-primary-anastomosis

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

16.3K Aufrufe.  University of Alberta. Ileocolic resection is commonly performed in several human diseases; however, little is known regarding the impact of intestinal resection on surgical illnesses. This article provides instruction on executing the procedure in mice with high success, providing a means to study the effects of ileocolic resection in models of disease.

Serie: Murine Ileocolic Bowel Resection with Primary Anastomosis

Small Bowel Transplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis.  In this article, we provide details of the technique to supplement the video.

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

Dünndarmtransplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

Forschung

Biologie

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer that often results in severe short bowel syndrome-like adverse effects including bile acid diarrhea, dehydration, electrolyte disturbances, and nutrient malabsorption. Here we introduce a murine ileal resection model, termed ileectomy, to evaluate tissue communication and the maintenance of systemic homeostasis. After ileal resection, circulating blood is permanently devoid of the ileum-specific endocrine hormone fibroblast growth factor 15 (FGF15), which releases its endocrinal inhibition of bile acid synthesis in the liver. In combination with the increased production and abolished reabsorption of bile acids after removing the ileum, mice that underwent surgery suffer from bile salt overaccumulation in the intestine and associated diarrhea, morbidity, and mortality. Novel usage of the surgery model introduced in this study may provide mechanistic and functional insights into ileal control of systemic metabolic regulation in physiology and disease.

Small intestine-dependent bile acid reabsorption and feedback inhibition of hepatic bile acid synthesis is important for systemic homeostasis and health. In this study, we describe a mouse model for ileal resection to evaluate ileectomy-induced bile malabsorption, overaccumulation, and toxicity in mouse intestine.

Overaccumulation Ileectomy-induzierten Galle im Darm der Maus

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

Medizin

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.

Murine distalen Kolostomie, ein neuartiges Modell Ablenkung Colitis bei C57BL/6 Mäusen

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

Creation of Colonic Anastomosis in Mice

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of colonic surgeries and can occur in up to 26% of surgical anastomoses, with mortality being up to 39% for patients with such a leak. Currently, there remains a paucity of data detailing the cellular mechanisms of anastomotic healing. Devising preventative strategies and treatment modalities for anastomotic leak could be greatly potentiated by a better understanding of appropriate anastomotic healing.
A murine model is ideal as previous studies have shown that the murine anastomosis is the most clinically similar to the human case as compared with other animal models. We offer an easily reproducible murine model of colonic anastomosis in mice that will allow for further illustration of anastomotic healing.

Anastomotic leak or breakdown after surgery is a major cause of postoperative morbidity and mortality. Our surgery for creating a colonic anastomosis is a reliable and reproducible method for studying the healing mechanisms of anastomoses.

Erstellung von Kolon Anastomose bei Mäusen

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of ...

Robotic Radical Cystectomy, Pelvic Lymph Node Dissection, and Intracorporeal Ileal Conduit Urinary Diversion

The robotic approach to radical cystectomy is compelling because of its oncologic equivalence to open radical cystectomy (ORC), its association with lower surgical blood loss, its potential association with shorter hospital stay after surgery. These factors suggest that the robotic approach to radical cystectomy may be an important component of enhanced recovery programs aimed at reducing surgical morbidity. This paper describes the importance of the cranial placement of robotic trocars, the use of Cadiere forceps for atraumatic bowel grasping, pelvic lymph node dissection (PLND), and utero-enteric anastomoses. Also discussed are steps that are critical for the successful outcome of RARC. In spite of the increased operating times and associated costs and the costs of robotic surgical platforms and equipment, adoption of the robotic technique by bladder cancer surgeons has increased. This paper describes a systematic and reproducible method that details robotic extended pelvic lymph node dissection, cystectomy/cystoprostatectomy, and intracorporeal ileal conduit urinary diversion.

This paper describes robotic radical cystectomy, pelvic lymph node dissection, and intracorporeal ileal conduit urinary diversion.

The robotic approach to radical cystectomy is compelling because of its oncologic equivalence to open radical cystectomy (ORC), its association with lower ...

<meta charset="utf8"></head><body>Mausmodell zur Untersuchung des postoperativen Ileus durch Darmmanipulation

Postoperative Ileus Murine Model

Most patients experience postoperative ileus (POI) after surgery, which is associated with increased morbidity, mortality, and hospitalization time. POI is a consequence of mechanical damage during surgery, resulting in disruption of motility in the gastrointestinal tract. The mechanisms of POI are related to aberrant neuronal sensitivity, impaired epithelial barrier function, and increased local inflammation. However, the details remain enigmatic. Therefore, experimental murine models are crucial for elucidating the pathophysiology and mechanism of POI injury and for the development of novel therapies.
Here, we introduce a murine model of POI generated via intestinal manipulation (IM) that is similar to clinical surgery; this is achieved by mechanical damage to the small intestine by massaging the abdomen 1-3 times with a cotton swab. IM delayed gastrointestinal transit 24 h after surgery, as assessed by FITC-dextran gavage and fluorescence detection of the segmental digestive tract. Moreover, tissue swelling of the submucosa and immune cell infiltration were investigated by hematoxylin and eosin staining and flow cytometry. Proper pressure of the IM and a hyperemic effect on the intestine are critical for the procedure. This murine model of POI can be utilized to study the mechanisms of intestinal damage and recovery after abdominal surgery.

<meta charset="utf8"></head><body>Postoperatives Ileus-Mausmodell

We describe a murine model of postoperative ileus generated via intestinal manipulation. Gastrointestinal transit function, pathologic changes, and immune cell activation were assessed 24 h after surgery.

Postoperatives Ileus-Mausmodell

Most patients experience postoperative ileus (POI) after surgery, which is associated with increased morbidity, mortality, and hospitalization time. POI ...

Immunologie und Infektion

Robot-assisted Total Mesorectal Excision and Lateral Pelvic Lymph Node Dissection for Locally Advanced Middle-low Rectal Cancer

Since their approval for clinical use, da Vinci surgical robots have shown great advantages in gastrointestinal surgical operations, especially in complex procedures. The high-quality 3-D visual, multijoint arm and natural tremor filtration allow the surgeon to expose and dissect more accurately with minimal invasion. Total mesorectal excision is the standard surgical technique for the treatment of resectable rectal cancer. To reduce the lateral recurrence rate, lateral pelvic lymph node dissection can be performed, as it is a safe and feasible procedure for locally advanced middle-low rectal cancer with a high possibility of metastasis to the lateral lymph nodes. However, the complexity of the anatomic structures and the high postoperative complication rate limit its application. Recently, several surgeons have increasingly used robotic techniques for total mesorectal excision and lateral pelvic lymph node dissection. Compared with open and laparoscopic surgery, the robotic technique has several advantages, such as less blood loss, fewer blood transfusions, minimal trauma, shorter postoperative hospitalization, and quicker recovery. A robotic approach is generally regarded as a reasonable alternative for complicated procedures such as lateral pelvic lymph node dissection, although there are a limited number of high-quality prospective randomized controlled studies reporting direct evidence. Here, we provide the detailed steps of robot-assisted total mesorectal excision and lateral pelvic lymph node dissection performed at the First Affiliated Hospital of Xi&#39;an Jiaotong University.

The robotic technique described herein aims to detail a stepwise approach to robot-assisted total mesorectal excision and lateral pelvic lymph node dissection for locally advanced (T3/T4) rectal cancer located below the peritoneal reflection.

Robotergestützte totale mesorektale Exzision und laterale Beckenlymphknotendissektion bei lokal fortgeschrittenem mittel-niedrigem Rektumkarzinom

Since their approval for clinical use, da Vinci surgical robots have shown great advantages in gastrointestinal surgical operations, especially in complex ...

Single-Anastomosis Duodeno-Ileal Bypass with Sleeve Gastrectomy Model in Mice

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis, cardiovascular events, and cancers) through restrictive and malabsorptive mechanisms. Understanding the mechanisms by which these procedures allow such improvements often require their transposition into animals, especially in mice, because of the ease of generating genetically modified animals. Recently, the single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) has emerged as a procedure that uses both restrictive and malabsorptive effects, which is being used as an alternative to gastric bypass in case of major obesity. Thus far, this procedure has been associated with strong metabolic improvements, which has led to a marked increase in its use in daily clinical practice. However, the mechanisms underlying these metabolic effects have been poorly studied as a result of a lack of animal models. In this article, we present a reliable and reproducible model of SADI-S in mice, with a special focus on perioperative management. The description and use of this new rodent model will be helpful for the scientific community to better understand the molecular, metabolic, and structural changes induced by the SADI-S and to better define the surgical indications for clinical practice.

Single-anastomosis duodeno-ileal bypass (SADI-S) is an emerging bariatric procedure with important metabolic effects. In this article, we present a reliable and reproducible model of SADI-S in mice.

Duodeno-ilealer Bypass mit Sleeve-Gastrektomie-Modell bei Mäusen

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes ...

Murine Ileocolische Darmresektion mit primärer Anastomose

Zusammenfassung

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Dünndarmtransplantation In Mice

Overaccumulation Ileectomy-induzierten Galle im Darm der Maus

Murine distalen Kolostomie, ein neuartiges Modell Ablenkung Colitis bei C57BL/6 Mäusen

Erstellung von Kolon Anastomose bei Mäusen

Robotic Radical Cystectomy, Pelvic Lymph Node Dissection, and Intracorporeal Ileal Conduit Urinary Diversion

Postoperatives Ileus-Mausmodell

Postoperatives Ileus-Mausmodell

Postoperatives Ileus-Mausmodell

Robotergestützte totale mesorektale Exzision und laterale Beckenlymphknotendissektion bei lokal fortgeschrittenem mittel-niedrigem Rektumkarzinom

Duodeno-ilealer Bypass mit Sleeve-Gastrektomie-Modell bei Mäusen