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. 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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. Technische Elemente der Operation zur Verhinderung Behinderung sind auf die Maximierung Lumendurchmesser an der Anastomose ab. Spateln des Ileums im Trennung Punkt erhöht den Lumendurchmesser, indem man den Darm in einer wie in Figur 1A dargestellten Winkel von 30 Grad. Bei der Aufnahme Bisse von Darmgewebe mit der Nähnadel, ist es unerlässlich, um die Ränder des Darms entrollen und übergeben Sie die Nadel etwa 0,5 mm von der Schnittkante. Zu große Bissen von Gewebe wird das Lumen einzugrenzen. Eine Option auf AVoid Lumeneinengung bei der Konstruktion der Anastomose ist, die Verwendung eines verdaulichen Stent einzusetzen, wie durch Kiernan et al. 15 beschrieben. Nicht-technische Aspekte der Obstruktion zu vermeiden, umfassen die Auswahl einer ausreichend kleinen Naht 4 und die Vermeidung von sperrigen Lumeninhalt durch die Fütterung von Tieren eine flüssige Diät einen Tag vor der Operation und 2 Tage nach der Operation. Die zweithäufigste Ursache für postoperative Mortalität ist Anastomoseninsuffizienz. Die Vermeidung dieser Komplikation erfordert die strikte Einhaltung der Grundsätze der Darmanastomose. Die erste ist eine ausreichende Blutversorgung. Nachdem die Gefäße Blut zum Bereich des Darms ist reseziert wurden ligiert, sollte das Darm visuell inspiziert vor Durchtrennung werden. Der ischämische Darmabschnitt wird düster erscheinen; dies sollte vollständig entfernt werden, wobei gesunde Kanten. Es ist wichtig, eine kleine Menge von Blutungen aus den Schnittkanten des verbleibenden Darms sichtbar zu machen. Eine wasserdichte Anastomosis ist auch wichtig, um das Auslaufen von luminalen Inhalte, die zu Sepsis zu vermeiden. Dies wird durch die Vermeidung von großen Lücken zwischen den Fäden, und sicherzustellen, dass alle Schichten des Darms im Stich enthalten (Figur 3) erreicht. Wundfestigkeit wird enorm in den ersten beiden postoperativen Tag vermindert, sind also richtig platziert Nähte wichtig, Stärke 16,17 bereitzustellen. Eine potenzielle Gefahr versehentlich nur einschließlich der Schleimhaut wie die Schleimhaut kann aus dem Lumen des Dickdarms durchtrennt verziehen. Dies kann durch Visualisierung der Schnittkante der Serosa und sanft reduziert prolapsed Schleimhaut wieder in das Lumen mit einer Pinzette zu vermeiden. Operativen Scheinkontrollen sollten in den experimentellen Studiendesign mit der ICR-Modell für die möglichen Verwechslung Wirkungen von chirurgischen Streß, Darm Manipulation und / oder Verlust der lleozökalklappe Steuerung berücksichtigt werden. Die Wahl des Schein-Verfahren wird auf der primären Frage abhängt, die in gefragtdie Studium. Zum Beispiel, wenn das Forschungsziel ist es, die Wirtsreaktion nach dem Verlust der lleozökalklappe bewerten, dann wäre es klug, eine Schein-OP-Gruppe, in der ein Darmanastomose durchgeführt, während die lleozökalklappe anstelle gehören. Dies wird typischerweise durch Durchschneiden in den Dünndarm ohne Resektion dann Durchführen einer Anastomose des Dünndarms in der gleichen Weise wie die Ileocolische Anastomose 7,11 geführt. Dies würde es Forschern, spezifisch auf die Auswirkungen der lleozökalklappe Verlust durch Steuern der Auswirkungen von Darmanastomose kommentieren. Alternativ, wenn das Ziel ist nur, die Auswirkungen der Darmresektion und Anastomose ohne zu kommentieren speziell auf Verlust des lleozökalklappe studieren, dann eine Scheinoperation sollte eine Laparotomie gehören, und der Darm sollte wie es für ICR behandelt werden, aber ohne Durchtrennung und Anastomose. Dies würde für die chirurgische Stress und Darmmanipulation steuern. Die im beschriebenen Verfahrenkann das Protokoll über leicht zur Durchführung Scheinverfahren angepasst werden. Dieses Protokoll beschreibt eine Ileocolische Resektion, die 1,5-2 cm terminalen Ileum und die Gesamtheit der Blinddarm entfernt, und imitiert eine Operation häufig bei Patienten mit Ileocolische Morbus Crohn durchgeführt. / - - Colitis-Modell, nach dem Mäuse entwickelten spontane Entzündung und Fibrose im Dünndarm analog zu Morbus Crohn Rezidiv 7,11 Ähnliche Verfahren wurden in der IL-10 verwendet. Die Prinzipien dieses Verfahrens kann auch auf andere Krankheitsmodelle anzuwenden. Durch Entfernen von 50% des Dünndarms mit Ileocolische Anastomose wurden Mausmodelle wurden verwendet, um die adaptive Darmwachstumsantwort in Kurzdarmsyndrom 4-6 untersucht. In der Zukunft kann dies auch ein nützliches Modell für die Untersuchung der systemischen und lokalen Auswirkungen der Chirurgie bei Darmkrebs zu werden.

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.

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			<td height="21" style="height:21px;width:216px;">LD101 liquid rodent diet</td>
			<td style="width:163px;">testdiet.com</td>
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			<td height="20" style="height:20px;">0.9% NaCl</td>
			<td>Baxter</td>
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			<td>Ziess</td>
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			<td>34-0483</td>
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			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Abbott&#160;</td>
			<td>05260-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass plate</td>
			<td></td>
			<td></td>
			<td>For operating surface</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotton swabs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro Castroviejo Needle holder, curved</td>
			<td>World Precision Instruments</td>
			<td align="right">503377</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo straight scissors</td>
			<td>World Precision Instruments</td>
			<td>555530S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissecting Scissors</td>
			<td>World Precision Instruments</td>
			<td align="right">15922</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dressing Forceps x 2</td>
			<td>World Precision Instruments</td>
			<td align="right">500363</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">5-0 silk pre-cut sutures</td>
			<td>Ethicon</td>
			<td>A182H</td>
			<td>For vessel ligation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">8-0 Prolene on BV130-5 needle</td>
			<td>Ethicon</td>
			<td>8732H</td>
			<td>For anastomosis</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">3-0 Silk on FS-2 needls</td>
			<td>Ethicon</td>
			<td>8665G</td>
			<td>For abdominal wall closure</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petroleum Jelly</td>
			<td>Vaseline</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 ml syringe</td>
			<td>BD biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone-iodine 7.5% surgical Scrub</td>
			<td><a href="http://betadine.com">betadine.com</a></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Heat lamps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">buprenorphine 0.3 mg/ml</td>
			<td>Reckitt Benckiser Healthcare Ltd.&#160;</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 wieder von postoperativen Tag 28 (Abbildung 2). Übertragbarkeit der Technik Ein neuer Student (BM) wurde gelehrt, die ICR Verfahren ausführen, um festzustellen, ob ein Betreiber ohne vorherige chirurgische Ausbildung könnte die Technik zu erlernen. Ausbildung begann mit dem Lernen, wie man die Einzelknopfnähten analog zu den in der Anastomose mit 8-0 Nylonnaht verwendet zuführen. Nähte wurden durchgeführt, um einen Einschnitt in einem Latex glo gemacht schließenve. Nachdem BM Lage, effizient zu platzieren Nähte durch den Handschuh, ohne zu reißen, beobachteten sie drei komplette Verfahren vor dem Versuch ein. Nachdem das erste Verfahren abgeschlossen war, sie dann beobachtet ein anderes Verfahren und versuchte zwei weitere. Von den drei Versuchstieren durchgeführt, die letzten beiden ging, um zu überleben und zu gedeihen. Das erste Tier wurde aufgrund von Komplikationen bei der Anastomose euthanasiert. <img alt="Figur 1" src="/files/ftp_upload/52106/52106fig1highres.jpg" /> Abbildung 1. Die Stufen der Ileocolische Resektion und Anastomose. (A) Ileozökalgegend Darm von Bauch geliefert. Schwarze Pfeile zeigen Gefäße ligiert werden, gestrichelten Linien zeigen Trennung Punkte für Doppelpunkt und terminalen Ileum. (B) Ileozökalgegend entfernt wird, werden Schnittenden Darmausgerichtet sind, den restlichen Blutversorgung wird durch schwarze Pfeile demonstriert. (C) Abgeschlossene Ileocolische Anastomosis. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52106/52106fig1large.jpg" target="_blank">Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen.</a> <img alt="Figur 2" src="/files/ftp_upload/52106/52106fig2highres.jpg" /> Abbildung 2. Tiergewichte an der Grundlinie, postoperativen Tag 14 und postoperativen Tag 28 keine signifikanten Veränderungen der Gewichte wurden nach dem Eingriff beobachtet, wie durch den Mann-Whitney-U statistische Tests bestimmt. ICR (Ileocolische Resektion-Gruppe), Control (Nichtoperative Kontrollen). <img alt="Figur 3" src="/files/ftp_upload/52106/52106fig3highres.jpg" /> Abbildung 3. Repräsentative Weg der Nähte durch Darmgewebe. H &amp; E gefärbten histologischen Schnitten von Ileum (rechts) und Doppelpunkt (links) aufgereiht sind, die Flugbahn des Fadens ne demonstrierenEdle durch die Gewebe (gestrichelte Linie) ab Punkt (a) bis zum Punkt (b). Der Knoten sollte am Punkt (c) angebunden werden. Beachten Sie, dass weniger Schleimhaut in der Anastomose als andere Gewebe enthalten. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52106/52106fig3large.jpg" target="_blank">Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen.</a>

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

Nanyang Technological University

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

The C-seal: A Biofragmentable Drain Protecting the Stapled Colorectal Anastomosis from Leakage

Colorectal anastomotic leakage (AL) is a serious complication in colorectal surgery leading to high morbidity and mortality rates1. The incidence of AL varies between 2.5 and 20% 2-5. Over the years, many strategies aimed at lowering the incidence of anastomotic leakage have been examined6, 7. 
The cause of AL is probably multifactorial. Etiological factors include insufficient arterial blood supply, tension on the anastomosis, hematoma and/or infection at the anastomotic site, and co-morbid factors of the patient as diabetes and atherosclerosis8. Furthermore, some anastomoses may be insufficient from the start due to technical failure.
Currently a new device is developed in our institute aimed at protecting the colorectal anastomosis and lowering the incidence of AL. This so called C-seal is a biofragmentable drain, which is stapled to the anastomosis with the circular stapler. It covers the luminal side of the colorectal anastomosis thereby preventing leakage.
The C-seal is a thin-walled tube-like drain, with an approximate diameter of 4 cm and an approximate length of 25 cm (figure 1). It is a tubular device composed of biodegradable polyurethane. Two flaps with adhesive tape are found at one end of the tube. These flaps are used to attach the C-seal to the anvil of the circular stapler, so that after the anastomosis is made the C-seal can be pulled through the anus. The C-seal remains in situ for at least 10 days. Thereafter it will lose strength and will degrade to be secreted from the body together with the gastrointestinal natural contents.
The C-seal does not prevent the formation of dehiscences. However, it prevents extravasation of faeces into the peritoneal cavity. This means that a gap at the anastomotic site does not lead to leakage.
Currently, a phase II study testing the C-seal in 35 patients undergoing (colo-)rectal resection with stapled anastomosis is recruiting. The C-seal can be used in both open procedures as well as laparoscopic procedures. The C-seal is only applied in stapled anastomoses within 15cm from the anal verge. In the video, application of the C-seal is shown in an open extended sigmoid resection in a patient suffering from diverticular disease with a stenotic colon.

The C-seal is a biofragmentable drain protecting the stapled colorectal anastomosis. In this video, details of the manufacturing, deployment and use are shown.

Die C-Dichtung: Ein Biofragmentable ablassen Schutz der Stapled kolorektalen Anastomose von Leakage

Colorectal anastomotic leakage (AL) is a serious complication in colorectal surgery leading to high morbidity and mortality rates1. The incidence of AL ...

Forschung

Medizin

Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

Orthotopen Dünndarm Transplantation in Ratten

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Application of End-to-end Anastomosis in Robotic Central Pancreatectomy

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of pancreatic body. With technological development, the robotic surgical system has shown its advantage in minimally invasive surgery and been increasingly applied in central pancreatectomy. However, reconstruction of the continuity of pancreas with end-to-end anastomosis after robotic central pancreatectomy has not been applied. In this study, we report surgical techniques for robotic central pancreatectomy with end-to-end anastomosis. The pancreas is reconstructed by duct-to-duct anastomosis of the pancreatic duct with a pancreatic stent inserted in the two stumps of pancreatic duct, and by end-to-end anastomosis of the pancreatic parenchyma. Compared with traditional central pancreatectomy with pancreaticoenteric anastomosis, this approach decreases the operative injury to the patient, and also conserves the integrity and continuity of the digestive duct and pancreatic duct. The robotic surgical system integrated with multiple instruments with flexible and precise movement is particularly suitable for the dissection and reconstruction of the pancreatic duct. We found that robotic central pancreatectomy with end-to-end anastomosis is safe and feasible, and we need more experience to evaluate its best indications and long-term outcomes.

The robotic central pancreatectomy with end-to-end anastomosis is feasible and safe for tumor in the pancreatic neck and proximal portion of pancreatic body. The operative techniques of this operation are presented.

Anwendung von End-to-End-Anastomose in Roboter-zentrale Pankreatektomie

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of ...

Endoscopic Septoplasty with Limited Two-line Resection: Minimally Invasive Surgery for Septal Deviation

Endoscopic septoplasty is a surgical procedure in otolaryngology that is commonly performed to treat nasal airway obstruction caused by nasal septal deviation. It has a long history with multiple variations. In this article, a modified endoscopic septoplasty procedure using the limited two-line resection (2LoRs) technique at the posterior and inferior junction of the cartilaginous and bony septum is presented based on embryologic and anatomic knowledge of the nasal septum and the biomechanics of cartilaginous behavior. With this procedure, the quadrangular cartilage can be preserved as much as possible, which is helpful in retaining the supporting framework and rigidness of the septum. 2LoRs has been proven effective and sound for the correction of nasal septal deviation with rare complications. This modified procedure can be applied to correct the deviated nasal septum in the absence of any external nasal deformity to improve nasal patency or to improve access to the middle meatus or to the axillary region of the middle turbinate. It may also be used to expand the indications of septoplasty to children and adolescents because of its minimally invasive approach.

Endoscopic septoplasty is a time-honored surgical procedure with multiple variations. This paper focuses on a step-by-step surgical approach to perform a modified septoplasty procedure known as limited two-line resection. This surgical technique can be applied to correct a deviated nasal septum in the absence of an external nasal deformity.

Endoskopische Septumplastik mit begrenzten zweizeilige Resektion: Minimal-Invasive Chirurgie für Septumdeviation

Endoscopic septoplasty is a surgical procedure in otolaryngology that is commonly performed to treat nasal airway obstruction caused by nasal septal ...

Vessel-sparing Excision and Primary Anastomosis

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short bulbar or posterior urethral strictures, urethroplasty by excision and primary anastomosis (EPA) is recommended. As EPA only requires the excision of the narrowed segment and the surrounding spongiofibrosis, a full-thickness transection of the corpus spongiosum, as performed in the traditional transecting EPA (tEPA), is usually unnecessary. Jordan et al. introduced the idea of a vessel-sparing approach in 2007, aiming to reduce surgical trauma, especially to the dual arterial blood supply of the urethra, and, thus, potentially reducing the risk of postoperative erectile dysfunction or glans ischemia. This approach could also be beneficial for subsequent urethral interventions such as redo urethroplasty using a free graft, in which a well-vascularized graft bed is imperative. Nevertheless, these potential benefits are only assumptions as prospective studies comparing the functional outcome of both techniques with validated questionnaires are currently lacking. Moreover, vessel-sparing EPA (vsEPA) should at least be able to provide similar surgical outcomes as tEPA. The aim of this paper is to give an elaborate, step-by-step overview of how to manage patients with isolated short bulbar or posterior urethral strictures with vsEPA. The main objective of this manuscript is to outline the surgical technique and to report the representative surgical outcome. A total of 117 patients were managed according to the described protocol. The analysis was performed on the entire patient cohort and on the bulbar (n = 91) and posterior (n = 26) vsEPA group separately. Success rates were 93.4% and 88.5% for the bulbar and posterior vsEPA, respectively. To conclude, vsEPA, as outlined in the protocol, provides excellent success rates with low complication rates for isolated short bulbar and posterior urethral strictures.

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

Schiff-schonende Exzision und primäre Anastomose

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

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 ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

Eine Anastomose Magenbypass (OAGB) bei Ratten

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

Studying Murine Small Bowel Mechanosensing of Luminal Particulates

Gastrointestinal (GI) motility is critical for normal digestion and absorption. In the small bowel, which absorbs nutrients, motility optimizes digestion and absorption. For this reason, some of the motility patterns in the small bowel include segmentation for mixing of luminal contents and peristalsis for their propulsion. Physical properties of luminal contents modulate the patterns of small bowel motility. The mechanical stimulation of GI mechanosensory circuits by transiting luminal contents and underlying gut motility initiate and modulate complex GI motor patterns. Yet, the mechanosensory mechanisms that drive this process remain poorly understood. This is primarily due to a lack of tools to dissect how the small bowel handles materials of different physical properties. To study how the small bowel handles particulates of varying sizes, we have modified an established in vivo method to determine small bowel transit. We gavage live mice with fluorescent liquid or tiny fluorescent beads. After 30 minutes, we dissect out the bowels to image the distribution of fluorescent contents across the entirety of the GI tract. In addition to high-resolution measurements of the geometric center, we use variable size binning and spectral analysis to determine how different materials affect small bowel transit. We have explored how a recently discovered "gut touch" mechanism affects small bowel motility using this approach.

To study how the small bowel handles particulates of varying sizes, we have modified an established in vivo method to determine small bowel transit.

Untersuchung der Mechanosensorik von luminalen Partikeln im murinen Dünndarm

Gastrointestinal (GI) motility is critical for normal digestion and absorption. In the small bowel, which absorbs nutrients, motility optimizes digestion ...

Modified Posterior Vertebral Column Resection for Patients with Thoracolumbar Kyphotic Deformity

Old compression vertebrae fracture or congenital kyphoscoliosis with abnormal vertebral body development and other diseases that invade the spine may cause severe thoracolumbar kyphotic deformity, often accompanied by intractable low back pain or compression of the spinal cord, leading to severe neurological symptoms or even paralysis. If conservative treatment cannot relieve the symptoms or correct the deformities, surgical treatment is usually needed. For severe kyphotic deformity, reconstruction of the physiological curvature and rigid fixation determine the prognosis of the patients. Osteotomy and orthopedics are the standard procedure for deformities with severe compression of the front and middle column, but the trauma to the patients is high, with a long operation time and massive blood loss. To avoid these disadvantages, we have developed a modified technique to remove the diseased vertebra unilaterally. In this technique, we use a modified trephine to resect the vertebral columns like in the pedicle screw technique by adding a locking instrument that can restrict the trephine to lower the risk of osteotomy and shorten the surgery time and blood loss.

We describe a modified technique for resecting the posterior vertebral column unilaterally based on a modified trephine for patients with thoracolumbar kyphotic deformity.

Modifizierte posteriore Wirbelsäulenresektion bei Patienten mit thorakolumbaler kyphotischer Deformität

Old compression vertebrae fracture or congenital kyphoscoliosis with abnormal vertebral body development and other diseases that invade the spine may ...

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 ...