This study was supported by a grant from the National R&amp;D Program for Cancer Control, Ministry of Health &amp; Welfare, Republic of Korea (1320270).

Ethik-Erklärung: Dieses Verfahren am Menschen wurde vom Institutional Review Board (IRB) an Ajou University Hospital genehmigt. 1. Vorbereitung für Chirurgie <ol><li> Lassen Sie eine weiche Kost einen Tag vor der Operation, mit nichts durch den Mund nach Mitternacht vor der Operation. Hinweis: Verwenden Sie keine Magensonde einsetzen, bevor und während der Operation. Dieses Protokoll verwendet keine Röhre um den Magen zu entleeren. Statt dass die intra-Magen-Gas kann durch Laparoskopie Punktionsnadel abgesaugt werden, wenn der Magen aufgetrieben wird eingeschränkte Sicht für die laparoskopische Chirurgie verursacht. </li><li> der Patient auf dem Operationstisch, zur Verwaltung der Induktionsmittel (Thiopental-Natrium, 100 mg) Nach der Platzierung. </li><li> Nach intravenöser Injektion eines Muskelrelaxans (Rocuroniumbromid, 1,0 mg / kg), legen Sie die Endotrachealtubus durch die Mundhöhle. </li><li> Starten Sie das Narkosegas (Sevofluran, 2 - 3%) über den Endotrachealtubus, dann überwachen the Patient für Stabilität während der Operation. </li></ol> 2. Chirurgische Verfahren vor intraoperative Gastros <ol><li> Der Patient wird in umgekehrter Trendelenburg-Position, und stehen auf der rechten Seite des Patienten. </li><li> Machen Sie 10 mm Schnitt am infraumbilical Bereich und legen Sie die erste Trokar in die Bauchhöhle. </li><li> Sobald der Trokar eingeführt wird, schließen Sie das Gasrohr für Kohlendioxid mit dem Trokar ein Pneumoperitoneum von 15 zu schaffen - 18 mm Hg. </li><li> Legen Sie eine starre 30 ° Laparoskop durch den Trokar, und erkunden Sie die Bauchhöhle auf einem Monitor mit dem laparoskopischen System verbunden. </li><li> Legen Sie 4 zusätzliche Trokare (ein 12 mm und drei 5 mm) als Arbeitsanschlüsse. Einsatz 12 mm Trokar an der rechten oberen Seite des Nabels. Legen Sie anderen drei 5 mm Trokare an der linken oberen Seite des Nabels und den beiden oberen äußeren Quadranten des Bauches. </li><li> Präparieren Sie die gastro-Band entlang der Querkolon in Richtung der inferioder Pol der Milz mittels Ultraschall-endoskopische Schere. </li><li> Ligieren der linken A. gastroepiploica mit Ursprung aus Milzgefäße an der Wurzel mit einer laparoskopischen Clip Vene und resect sie Ultraschall endoskopische Schere nach der Belichtung der oberen Grenze des Pankreasschwanz. </li><li> Mit Ultraschall-endoskopische Schere, sezieren die peripyloric Lymphknoten rund um den Pförtner und dem Kopf der Bauchspeicheldrüse, und die gastroepiploica Schiffe auf ihren Ursprung resezieren. </li><li> Vor Resektion des ersten Teils des Duodenums, klemmen das Duodenum mit einem laparoskopischen Klammer zu Migration von Gas in den Dünndarm während des gastroskopischen Verfahren verhindern (Abbildung 1). </li></ol> 3. Die intraoperative Gastros <ol><li> Bewegen Sie auf der linken Seite des Kopfes des Patienten intraoperative Gastroskopie durchzuführen. </li><li> Setzen Sie einen Mundschutz, die Zähne und das Zahnfleisch während der intraoperative Gastroskopie zu schützen. </li><li> Drehen Sie den head des Patienten auf der linken Seite zum Einführen des Gastroskop. </li><li> Legen Sie die Gastroskop in den Magen vorsichtig durch den Mund und die Speiseröhre, eine Schädigung der Schleimhaut der Mundhöhle, die Speiseröhre und den Magen zu vermeiden. </li><li> Insufflate den Magen durch Gas durch das Gastroskop Injizieren der primären Läsion zu lokalisieren. Hinweis: Die meisten Gastroskopen eine bestimmte Taste haben Gas in den Darm während des Verfahrens zu injizieren. </li><li> Ungefähre den Abstand zwischen dem proximalen Rand des Tumors und der gastroösophagealen Übergang mit Hilfe der Skala auf dem Gastroskop markiert (Abbildung 2), und führen Sie den gesamten Magen (wie zuvor beschrieben) , wenn der Abstand zu kurz ist , den proximalen Magen zu retten. 10 </li><li> Wenn Gastrektomie nicht notwendig ist, legen Sie eine endoskopische Injektor durch ein kleines Loch des Gastroskop und injizieren 2 - 3 ccm Indigokarmin in die Magenwand eine endoskopische Injektor in einem Bereich 2 mit - 3 cm proximal vom Tumorrand, Vermeidung von intra- oder extraluminaler Leckage (3A). <ol><li> Führen Sie die Nadel des Injektors in die Magenwand schräg die Perforation der Magenwand von der Nadel zu vermeiden. Darüber hinaus injizieren Indigokarmin sehr langsam intraluminale Leckage während der Injektion zu identifizieren. </li></ol></li><li> Nach dem Farbstoff Injektion zu aspirieren Füllgas durch das Gastroskop das laparoskopische Verfahren zu erleichtern. Hinweis: Die meisten Gastroskopen eine bestimmte Taste haben Füllgas während des Verfahrens zu aspirieren. </li></ol> 4. Verfahren nach intraoperative Gastros <ol><li> Rückkehr zur rechten Seite des Patienten nach der Gastroskopie Verfahren. </li><li> In der laparoskopischen Ansicht, bestätigen den proximalen Teil des Tumors durch serosal Färbung durch blauen Farbstoff (3B) zu identifizieren. </li><li> Markieren Sie den gefärbten Teil werden mit einem endoskopischen Clip reseziert, die Vermeidung Beteiligung der Tumor an der WiederAbschnitt Marge (4A). Anmerkung: Wir empfehlen, dieses Verfahren innerhalb von 5 - 10 min nach der Operation beginnt, da der Farbstoff manchmal während der nachfolgenden Verfahren ausgewaschen werden kann. </li><li> Wenn der Farbstoff nicht in der Serosa detektiert werden kann, sofort wieder einzuspritzen, den Farbstoff unter gastroskopischen Visualisierung. </li></ol> 5. Verfahren nach intraoperative Gastros <ol><li> Reseziert Duodenum und führen auch andere Verfahren zur Lympknotendissektion nach den japanischen Behandlungsrichtlinien. 11 </li><li> Nach Lympknotendissektion, den Magen an der richtigen Stelle mindestens 3 cm proximal von der Tumorrand reseziert, durch den Markierungsklipp geführt, unter Verwendung von 2 endoskopische Heftern. </li><li> Sobald das distale Magen reseziert, um die Probe in einem endoskopischen Tasche einsetzen und aus der Bauchhöhle durch ein 2 zurückziehen - cm Verlängerung der infraumbilical Trokar Website 3. </li><li> Senden Sie das Gewebe aus dem proximalen Rand zuPathologie für die intraoperative histologische Auswertung. </li><li> Nach Bestätigung der tumorfreien Gewebes in dem proximalen Abschnitt des resezierten Magen, eine Anastomose zwischen dem restlichen Magen und dem proximalen Jejunum mit intrakorporalen linear staplers (4B) durchführen. </li></ol>

<ol>
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Y., 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=Is+totally+laparoscopic+gastrectomy+less+invasive+than+laparoscopy-assisted+gastrectomy?:+prospective,+multicenter+study.">Is totally laparoscopic gastrectomy less invasive than laparoscopy-assisted gastrectomy?: prospective, multicenter study.</a> J Gastrointest Surg. 12, 1015-1021 (2008).</li><li>Hyung, W. J., 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=Intraoperative+tumor+localization+using+laparoscopic+ultrasonography+in+laparoscopic-assisted+gastrectomy.">Intraoperative tumor localization using laparoscopic ultrasonography in laparoscopic-assisted gastrectomy.</a> Surg Endosc. 19, 1353-1357 (2005).</li><li>Jeong, O., Cho, S. B., Joo, Y. E., Ryu, S. Y., Park, Y. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Training+of+surgical+endoscopists+in+Korea:+assessment+of+the+learning+curve+using+a+cumulative+sum+model.">Training of surgical endoscopists in Korea: assessment of the learning curve using a cumulative sum model.</a> J Surg Educ. 69, 559-563 (2012).</li><li>Cho, W. Y., 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=Hybrid+natural+orifice+transluminal+endoscopic+surgery:+endoscopic+full-thickness+resection+of+early+gastric+cancer+and+laparoscopic+regional+lymph+node+dissection--14+human+cases.">Hybrid natural orifice transluminal endoscopic surgery: endoscopic full-thickness resection of early gastric cancer and laparoscopic regional lymph node dissection--14 human cases.</a> Endoscopy. 43, 134-139 (2011).</li><li>Abe, N., 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=Endoscopic+full-thickness+resection+with+laparoscopic+assistance+as+hybrid+NOTES+for+gastric+submucosal+tumor.">Endoscopic full-thickness resection with laparoscopic assistance as hybrid NOTES for gastric submucosal tumor.</a> Surg Endosc. 23, 1908-1913 (2009).</li><li>Hoetker, M. 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The authors have no conflicts of interest or financial ties to disclose.

In diesem Protokoll haben wir ein Verfahren für die intraoperative Gastroskopie, um genau und zuverlässig Tumorlokalisation in dem mittleren Drittel des Magens identifizieren. Wir konnten erfolgreich distale subtotale Gastrektomie über die Ergebnisse der intraoperativen Gastros Basis durchführen. Bis heute wurden verschiedene Verfahren angewandt, um die Lage der frühen Magenkrebs in der laparoskopischen Ansicht zu identifizieren. Die häufigsten Methoden der präoperativen gastroskopisc...

Die laparoskopische Chirurgie hat in der Behandlung der Wahl für Magenfrühkarzinomen ostasiatischen Ländern geworden, darunter Korea und Japan. 1 Der Vorteil dieser Operation wurde in mehreren klinischen Studien für Magenfrühkarzinomen (EGC) gut demonstriert. 2-4 Die meisten die Verfahren in diesen Studien wurden durch Laparoskopie durchgeführt, aber die Identifizierung des Tumorlokalisation, Resektion des Magens, einschließlich des Primärtumors und Rekonstruktion wurden via minilaparotomy durchgeführt. Daher wurde minilaparotomy hat Chirurgie erfordert Bezeichnung &quot;gastrectomy Laparoskopie-assistierte.&quot; Kürzlich haben jedoch chirurgische Verfahren entwickelt Trauma zu minimieren, für Patienten, die die postoperativen Verbesserung der Lebensqualität. Da dieses Konzept auch in der laparoskopischen Chirurgie für Magenkrebs, einige erfahrene laparoskopischen Chirurgen minilaparotomy zu vermeiden, haben angewendet wurde versucht. Völlig laparoskopische Gastrektomie (TLG) für gastric Krebs erfordert, dass alle Verfahren nur laparoskopischen Geräten vorgenommen verwenden, ohne dass eine zusätzliche minilaparotomy für die Proben Resektion und Anastomose erfordern. Da diese Art der Operation führt zu weniger Schmerzen und eine schnellere Erholung relativ zu öffnen oder Laparoskopie-assistierte Chirurgie, die Laparotomie benötigen, bevorzugen es mehr Chirurgen. 5,6 jedoch ein Hindernis für die TLG für Magenkrebs ist Tumorlokalisation ohne direkte Visualisierung oder Palpation den Bereich der Magen-Resektion zu bestimmen. Tumor Abwesenheit am Resektionsrand ist sehr wichtig, den Erfolg bei Magenkrebs Operation zu erreichen. Bei Tumorbeteiligung an der Resektionsrand während distalen Gastrektomie für Magenkrebs ist, ist eine zusätzliche Resektion benötigt in den verbleibenden Magen verlassen Tumor zu vermeiden. In offenen und Laparoskopie-assistierte Chirurgie, kann der primäre Tumor leicht durch Palpation oder direkte Visualisierung durch eine temporäre Gastrostomie lokalisiert werden. Da jedoch die Primary Tumor ist in der laparoskopischen Ansicht, die Bestimmung Resektionsrändern für EGC in TLG schwierig sein kann nicht leicht erkannt. Mehrere andere Verfahren zusätzliche präoperative Gastroskopie erfordern Resektionsrändern zu bestimmen , wurden vorgeschlagen. 7-9 jedoch weitere präoperative Gastroskopie kann für die Patienten unbequem sein. Wir führten ein Verfahren für die intraoperative Gastroskopie direkt Tumoren während TLG für Magenkrebs im mittleren Drittel des Magens zu markieren. In diesem Protokoll verwendeten wir die laparoskopische Chirurgie für Patienten mit frühen Magenkrebs bei den präoperativen Untersuchungen, die in absoluter Indikation sind nicht für die endoskopische Submukosa-Dissektion enthalten.

<table border="0" cellpadding="0" cellspacing="0" width="717">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Pentothal Sodium</td>
			<td style="width:157px;">JW pharmatheutical Co.</td>
			<td style="width:119px;">644912100</td>
			<td style="width:225px;">It was used as induction asgent</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Rocumeron</td>
			<td style="width:157px;">&#160;Ilsung pharmatheutical Co.</td>
			<td style="width:119px;">655402960</td>
			<td style="width:225px;">It was used for general anesthestia as muscle relaxant.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">SevoFRAN</td>
			<td style="width:157px;">Hana pharmatheutical Co.</td>
			<td style="width:119px;">657801060</td>
			<td style="width:225px;">It was used as general anesthetics</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Endoscopy system</td>
			<td style="width:157px;">Olympus Corp.</td>
			<td style="width:119px;">CLV-260</td>
			<td style="width:225px;">It was used for intraoperative GFS</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Laparoscopic surgery system</td>
			<td style="width:157px;">Striker Endoscopy</td>
			<td style="width:119px;">1488 HD 3-CHIP</td>
			<td style="width:225px;">It was a system for laparoscopic surgery</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Ultrasonic scissor</td>
			<td style="width:157px;">Johnson and Johnson Medical Corp.</td>
			<td style="width:119px;">HAR23</td>
			<td style="width:225px;">It was used during tissue dissection</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Laparoscopic clip</td>
			<td style="width:157px;">Johnson and Johnson Medical Corp.</td>
			<td style="width:119px;">ER420</td>
			<td style="width:225px;">It was used for ligation of the vessels.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Indigo carmine</td>
			<td style="width:157px;">Korea United Pharma</td>
			<td style="width:119px;">Carmine</td>
			<td style="width:225px;">It was injected into the gastric wall</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:216px;">Linear stapler</td>
			<td style="width:157px;">Johnson and Johnson Medical Corp.</td>
			<td style="width:119px;">ECHELON FLEX Powered Endopath Stapler</td>
			<td style="width:225px;">It was used for resection the stomach</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Gastroscopic injector</td>
			<td style="width:157px;">TaeWoong Medical</td>
			<td style="width:119px;">Cobra injector</td>
			<td style="width:225px;">I was used for gastroscopic injection of blue dye</td>
		</tr>
	</tbody>
</table>

intraoperative gastroscopy for tumor localization in laparoscopic surgery for gastric adenocarcinoma

Bestimmen Resektionsrändern für Magenkrebs, die auf die Serosa-Oberfläche exponiert sind nicht im Magen, ist das wichtigste Verfahren in völlig laparoskopischen Gastrektomie (TLG). Ziel dieses Protokolls ist es, ein Verfahren für die intraoperative Gastroskopie, einzuführen, um direkt Tumoren während TLG für Magenkrebs im mittleren Drittel des Magens markieren. Patienten, die mit einem Adenokarzinom im mittleren Drittel des Magens diagnostiziert wurden, wurden in diesem Fall Serie eingeschrieben. Vor der Operation wird eine zusätzliche Gastroskopie für Tumorlokalisation nicht durchgeführt. Unter Vollnarkose wird laparoskopische Mobilisierung des Magens zuerst ausgeführt. Nachdem der erste Abschnitt des Duodenums wird aus der Bauchspeicheldrüse mobilisiert und eingespannt, der Chirurg bewegt sich zu der anderen Seite für die gastroskopischen Verfahren. Auf das Einführen eines Gastroskop durch die Mundhöhle in den Magen, 2 - 3 ml Indigokarmin wird über einen endoskopischen Injektor in den Magen verabreicht muscle Schicht am proximalen Rand des Magens. Die Lage von gefärbten serosa in der laparoskopischen Ansicht wird verwendet distale subtotale Gastrektomie zu führen, jedoch Gastrektomie durchgeführt wird, wenn der Tumor an der gastroösophagealen Übergang zu nahe ist. Eine Probe wird nach distalen Gastrektomie abgetastet, um eine ausreichende Länge von Resektionsrandes auf Tumor vor der Rekonstruktion bestätigen. In unserem Fall Serie hatten alle Patienten tumorfrei Margen und benötigt keine zusätzliche Resektion. Es gab keine Morbidität zur Gastroskopie Verfahren bezogen, und die Zeit für das Verfahren erforderlich ist allmählich auf etwa 5 Minuten verringert. Die intraoperative Gastroskopie für Tumorlokalisation ist eine genaue und tolerierte Methode für Patienten mit Magenkrebs total laparoskopische distalen Gastrektomie unterzogen.

Von 20 Patienten, die intraoperative Gastroskopie unterzogen, unterzog 18 distale subtotale Gastrektomie, durch die Ergebnisse bei der Gastroskopie geführt. Allerdings führten wir den gesamten Magen auf zwei Patienten, weil das proximale Tumorrand an der gastroösophagealen Übergang zu nahe war, als auf die intraoperative Gastroskopie bestimmt. Der Abstand vom Tumor zum proximalen Resektionsrand betrug 3,5 cm und 2,5 cm in diesen beiden Patienten (Tabelle 1). <p c...

Watch this Scientific Journal Video about Intraoperative Gastroscopy for Tumor Localization in Laparoscopic Surgery for Gastric Adenocarcinoma at JoVE.com

Intraoperative Gastroscopy for Tumor Localization in Laparoscopic Surgery for Gastric Adenocarcinoma

In early gastric cancer, the aim of surgery is to precisely remove the distal stomach including the primary tumor. To do this, accurate localization of the tumor is crucial, especially in totally laparoscopic surgery. This protocol describes a procedure for intraoperative gastroscopy in totally laparoscopic subtotal gastrectomy.

Die intraoperative Gastroskopie für Tumorlokalisation in der laparoskopischen Chirurgie für Adenokarzinom des Magens

Determining resection margins for gastric cancer, which are not exposed to the serosal surface of the stomach, is the most important procedure during ...

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12.6K Aufrufe.  Ajou University, School of Medicine. In early gastric cancer, the aim of surgery is to precisely remove the distal stomach including the primary tumor. To do this, accurate localization of the tumor is crucial, especially in totally laparoscopic surgery. This protocol describes a procedure for intraoperative gastroscopy in totally laparoscopic subtotal gastrectomy.

Serie: Intraoperative Gastroscopy for Tumor Localization in Laparoscopic Surgery for Gastric Adenocarcinoma

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

Roux-en-Y-Magenbypass Operation in Ratten

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

Forschung

Medizin

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

Screening für Melanom Modifiers mit einem Zebrafisch Autochthone Tumormodell

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

Assessing Specificity of Anticancer Drugs In Vitro

A procedure for assessing specificity of anticancer drugs in vitro using cultures containing both tumor and non-tumor cells is demonstrated. The key element is the quantitative determination of a tumor-specific genetic alteration in relation to a universal sequence using a dual-probe digital PCR assay and the subsequent calculation of the proportion of tumor cells. The assay is carried out on a culture containing tumor cells of an established line and spiked-in non-tumor cells. The mixed culture is treated with a test drug at various concentrations. After the treatment, DNA is prepared directly from the survived adhesive cells in wells of 96-well plates using a simple and inexpensive method, and subjected to a dual-probe digital PCR assay for measuring a tumor-specific genetic alteration and a reference universal sequence. In the present demonstration, a heterozygous deletion of the NF1 gene is used as the tumor-specific genetic alteration and a RPP30 gene as the reference gene. Using the ratio NF1/RPP30, the proportion of tumor cells was calculated. Since the dose-dependent change of the proportion of tumor cells provides an in vitro indication for specificity of the drug, this genetic and cell-based in vitro assay will likely have application potential in drug discovery. Furthermore, for personalized cancer-care, this genetic- and cell-based tool may contribute to optimizing adjuvant chemotherapy by means of testing efficacy and specificity of candidate drugs using primary cultures of individual tumors.

Assessing Specificity of Anticancer Drugs In Vitro

The goal of this protocol is to assess specificity of anticancer drugs in vitro using mixed cultures containing both tumor and non-tumor cells.

Die Beurteilung Spezifität von Anti-Krebs-Medikamente<em&gt; In Vitro</em

A procedure for assessing specificity of anticancer drugs in vitro using cultures containing both tumor and non-tumor cells is demonstrated. The key ...

Cone Beam Intraoperative Computed Tomography-based Image Guidance for Minimally Invasive Transforaminal Interbody Fusion

Transforaminal lumbar interbody fusion (TLIF) is commonly used for the treatment of spinal stenosis, degenerative disc disease, and spondylolisthesis. Minimally invasive surgery (MIS) approaches have been applied to this technique with an associated decrease in estimated blood loss (EBL), length of hospital stay, and infection rates, while preserving outcomes with traditional open surgery. Previous MIS TLIF techniques involve significant fluoroscopy that subjects the patient, surgeon, and operating room staff to non-trivial levels of radiation exposure, particularly for complex multi-level procedures. We present a technique that utilizes an intraoperative computed tomography (CT) scan to aid in placement of pedicle screws, followed by traditional fluoroscopy for confirmation of cage placement. Patients are positioned in the standard fashion and a reference arc is placed in the posterior superior iliac spine (PSIS) followed by intraoperative CT scan. This allows for image-guidance-based placement of pedicle screws through a one-inch skin incision on each side. Unlike traditional MIS-TLIF that requires significant fluoroscopic imaging during this stage, the operation can now be performed without any additional radiation exposure to the patient or operating room staff. After completion of the facetectomy and discectomy, final TLIF cage placement is confirmed with fluoroscopy. This technique has the potential to decrease operative time and minimize total radiation exposure.

The purpose of this article is to provide image-guidance for minimally invasive transforaminal interbody fusion.

Kegelstrahl Intraoperative Computertomographie-basierte Bildführung für minimalinvasive transforaminale Interbody-Fusion

Transforaminal lumbar interbody fusion (TLIF) is commonly used for the treatment of spinal stenosis, degenerative disc disease, and spondylolisthesis. ...

Intraoperative Ultrasound in Spinal Surgery

Since the 1980s, there have been several reports for the use of intraoperative ultrasound as a useful adjunct in spinal surgery. However, with the advent of newer cutting-edge imaging modalities, the use of intraoperative ultrasound in spine surgery has largely fallen out of favor. Despite this, intraoperative ultrasound continues to provide several advantages over other intraoperative techniques such as magnetic resonance imaging and computed tomography including being more cost-effective, efficient, and easy to operate and interpret. Additionally, it remains the only method for the real-time visualization of soft tissue and pathologies. This paper focuses on the advantages of using intraoperative ultrasound, especially in cases of intradural lesions and lesions ventral to the thecal sac when approaching posteriorly.

Here, we present a protocol on the use of intraoperative ultrasound in spinal surgery, particularly in cases of intradural lesions and lesions in the ventral spinal canal when using a posterior approach.

Intraoperativer Ultraschall in der Wirbelsäulenchirurgie

Since the 1980s, there have been several reports for the use of intraoperative ultrasound as a useful adjunct in spinal surgery. However, with the advent ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or excision. Many of these nodules are identified and excised via minimally invasive thoracic surgery; however, subcentimeter and subsolid nodules are frequently difficult to identify intra-operatively. This can be mitigated by the use of electromagnetic transthoracic needle localization. This protocol delineates the step-by-step process of electromagnetic localization from the pre-operative period to the postoperative period and is an adaptation of the electromagnetically guided percutaneous biopsy previously described by Arias et al. Pre-operative steps include obtaining a same day CT followed by the generation of a three-dimensional virtual map of the lung. From this map, the target lesion(s) and an entry site are chosen. In the operating room, the virtual reconstruction of the lung is then calibrated with the patient and the electromagnetic navigation platform. The patient is then sedated, intubated, and placed in the lateral decubitus position. Using a sterile technique and visualization from multiple views, the needle is inserted into the chest wall at the prechosen skin entry site and driven down to the target lesion. Dye is then injected into the lesion and, then, continuously during needle withdrawal, creating a tract for visualization intra-operatively. This method has many potential benefits when compared to the CT-guided localization, including a decreased radiation exposure and decreased time between the dye injection and the surgery. Dye diffusion from the pathway occurs over time, thereby limiting intra-operative nodule identification. By decreasing the time to surgery, there is a decrease in wait time for the patient, and less time for dye diffusion to occur, resulting in an improvement in nodule localization. When compared to electromagnetic bronchoscopy, airway architecture is no longer a limitation as the target nodule is accessed via a transparenchymal approach. Details of this procedure are described in a step-by-step fashion.

Presented here is a protocol for lung nodule localization using dye marking via electromagnetically navigated transthoracic needle access. The technique described here can be accomplished in the peri-operative period to optimize nodule localization and to successful resection when performing minimally invasive thoracic surgery.

Elektromagnetische Navigation Transthorakale Knotenlokalisation für die minimalinvasive Thoraxchirurgie

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or ...

Laparoscopic Radical Left Pancreatectomy for Pancreatic Cancer: Surgical Strategy and Technique Video

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological resection of left-sided pancreatic cancer. Several surgical techniques have been described in recent years, but few were specifically designed for minimally invasive approaches. This study describes and demonstrates a standardized and reproducible technique for an adequate oncological resection of pancreatic cancer: laparoscopic radical left pancreatectomy (LRLP).
A 61-year-old woman presented with an incidental finding of a 3 cm mass in the left pancreas suspect for malignancy. Imaging did not reveal distant metastases, central vascular involvement, or morbid obesity, hence the patient was suitable for LRLP. This study describes the main steps of LRLP for pancreatic cancer. First, the lesser sac is opened by transecting the gastrocolic ligament. The splenic flexure of the colon is mobilized and the inferior border of the pancreas including Gerota&#39;s fascia is dissected down to the inferior border of the spleen. The pancreas is tunneled and hung, including Gerota&#8217;s fascia with a vessel loop. At the pancreatic neck, a tunnel is created between the pancreas and the portal vein, likewise a vessel loop is passed. The pancreas is then transected using the graded compression technique with an endostapler. Both the splenic vein and artery are transected before completing the resection. The entire specimen is extracted in a retrieval bag via a small Pfannenstiel incision.
Duration of the surgery was 210 min with 250 mL blood loss. Pathology revealed a R0-resection (&#62;1 mm) of a well-to-moderately differentiated adenocarcinoma originating from an intraductal papillary mucinous neoplasm. A total of 15 tumor-negative lymph nodes were resected. This is a detailed description of LRLP for left-sided pancreatic cancer as is currently being used within the international, multicenter randomized DIPLOMA (Distal Pancreatectomy Minimally Invasive or Open for PDAC) trial.

Oncologically safe left pancreatectomy requires radical resection (R0), Gerota&#8217;s (perirenal) fascia resection, and adequate lymph node dissection. This study describes the technical details of laparoscopic radical left pancreatectomy (LRLP), used in the first international multicenter randomized trial comparing minimally invasive with open left pancreatectomy for pancreatic cancer, the DIPLOMA trial.

Laparoskopische radikale linke Panacreatektomie für Bauchspeicheldrüsenkrebs: Chirurgische Strategie und Technik Video

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological ...

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

Ein Protokoll für Roux-en-Y Magenbypass bei Ratten mit linearen Heftern

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Laparoscopy-endoscopy Cooperative Surgery for the Treatment of Gastric Gastrointestinal Stromal Tumors

In view of the shortcomings of endoscopic or laparoscopic surgeries alone in the treatment of gastric gastrointestinal stromal tumors (G-GISTs), this approach makes an innovative improvement in the treatment of G-GISTs that are less than 5 cm in size. Laparoscopy-endoscopy cooperative surgery (LECS) is used to combine endoscopic and laparoscopic surgeries, fully realizing their respective advantages and avoiding their drawbacks. The main steps are as follows. First, gastroscopy and laparoscopy are combined to confirm the location and boundary of the tumor. Tumor resection is carried out laparoscopically, guided by a gastroscope. The specimen is removed orally and the gastric wound closed laparoscopically. Then, gastroscopy and laparoscopy are combined to determine whether there is wound bleeding, if the suture is satisfactory, and if the gastric cavity is deformed.
LECS has natural advantages in the treatment of G-GISTs that are less than 5 cm in size. The accurate estimation of tumor location and boundary greatly improves the complete resection rate of tumors. The risk of tumor rupture is substantially reduced, and the long-term prognosis of patients is significantly improved. The process allows for accurate resection of the tumor, maximum preservation of normal gastric tissue and organ function, and avoids postoperative gastric deformation. The patient's postoperative rehabilitation is greatly accelerated, and oral feeding can resume on the day of the operation. The specimen is taken out through the mouth to avoid the need for an extended abdominal incision. This greatly reduces the patient's postoperative pain and scarring. The method greatly shortens the postoperative hospital stay (i.e., discharge is possible on the day after the operation), increasing the turnover of hospital beds.

Laparoscopy-endoscopy cooperative surgery is appropriate for the treatment of gastric gastrointestinal stromal tumors less than 5 cm in size. It can achieve the respective advantages of endoscopic and laparoscopic surgeries while avoiding their drawbacks.

Laparoskopie-Endoskopie Kooperative Chirurgie zur Behandlung von Magen-Darm-Stromatumoren

In view of the shortcomings of endoscopic or laparoscopic surgeries alone in the treatment of gastric gastrointestinal stromal tumors (G-GISTs), this ...

Fluorescent Laparoscopic Central Hepatectomy for Liver Cancer Using Indocyanine Green Negative Staining

Laparoscopic hepatectomy is an important treatment method for liver cancer. In the past, the resection boundary was usually determined by intraoperative ultrasound, important vascular structures, and surgeon experience. With the development of anatomical hepatectomy, visual surgery technology has gradually been applied to this type of surgery, particularly indocyanine green (ICG)-guided anatomical hepatectomy. As ICG can be specifically ingested by hepatocytes and used for fluorescence tracing, negative staining techniques have been applied according to different tumor positions. Under ICG fluorescent guidance, the surface boundary and deep resection plane can be more accurately displayed during liver resection. Thus, the tumor-bearing liver segment can be anatomically removed, which helps to avoid damage to important vessels and reduce ischemia or congestion of the remaining liver tissue. Finally, the incidence of postoperative biliary fistula and liver dysfunction is reduced; therefore, a better prognosis is obtained after the resection of liver cancer. Centrally located liver cancer is usually defined as a tumor located at segments 4, 5, or 8 that requires resection of the middle section of the liver. These are among the most difficult hepatectomies to perform because of the large surgical wounds and multiple vessel transections. Based on the specific tumor location, we formulated the required resection ranges by designing personalized fluorescent staining strategies. By completing anatomical resection based on the portal territory, this work aims to achieve the best therapeutic effect.

The present protocol describes fluorescent negative staining in laparoscopic central hepatectomy. This technique can make hepatectomy more accurate and precise.

Fluoreszierende laparoskopische zentrale Hepatektomie bei Leberkrebs mit Indocyaningrün-Negativfärbung

Laparoscopic hepatectomy is an important treatment method for liver cancer. In the past, the resection boundary was usually determined by intraoperative ...