The authors have no acknowledgments.

This study was permitted by the Ethics Committee of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine.
1. Patient selection
<ol>
	<li>Advise the patients suspected of suffering pancreatic ductal adenocarcinoma (PDAC) in the pancreas head to take a contrast-enhanced computed tomography (CT) scan at first. Then select the following patients: borderline resectable cases after neoadjuvant therapy, tumor regresses significantly and has the possibility of radical resection; meanwhile, SMV reconstruction is potentially needed.</li>
	<li>Exclude the following patients: (1) metastatic PDAC; (2) patients with a poor general condition that cannot tolerate major surgery; (3) vascular involvement of SMA, common hepatic artery (CHA), or celiac artery (CA).</li>
</ol>
2. Surgical technique
<ol>
	<li>Operative setting
	<ol>
		<li>Place the anesthetized patient in a completely supine position, with legs spread out.</li>
		<li>During the sterile exposition, ensure that the suprapubic region is exposed adequately for the later specimen removal by a Pfannenstiel incision. Ensure that the operator surgeon stands on the right side, the first assistant is on the left side, and the second&#160;assistant holding the laparoscope is positioned between the patient&#39;s legs. Use the 5-port technique to perform the procedure (Figure 1).</li>
		<li>After the pneumoperitoneum is created, introduce the rigid laparoscope through the sub-umbilical 12 mm trocar, and place the other four trocars along a semi-circle.</li>
	</ol>
	</li>
	<li>Exploration phase
	<ol>
		<li>Explore the intraperitoneal organs and peritoneal surfaces meticulously for unexpected extrapancreatic metastases.</li>
		<li>Resect the greater omentum and open the lesser sac by dividing the gastrocolic ligament.</li>
		<li>Ligate Henle&#39;s gastrocolic trunk vein. Explore the gap between the pancreatic neck and SMV at the lower edge of the pancreas. 
		NOTE: The tunnel between the pancreatic neck and SMV can not be created easily if the anterior wall of SMV is involved.</li>
	</ol>
	</li>
	<li>Dissection phase:
	<ol>
		<li>Subcolonic mesenteric approach: For this approach, follow steps 2.3.2-2.3.5.</li>
		<li>After the transverse colon and its mesentery are elevated cephalad, place the entire small bowel on the left side to facilitate exposure of the pancreaticoduodenal region.</li>
		<li>Expose the second and third parts of the duodenum after the mesentery section.</li>
		<li>In order to reconfirm resectability and mobilize the posterior of the pancreatic head, expose the inferior vena cava (IVC), left renal vein (LRV), celiac trunk, aorta, and SMA (Figure 2).</li>
		<li>Then perform the dissection along the SMV to clear all the tissues on the right side. In order to expose the SMV fully, ligate the root of the middle colonic artery after confirming that there is no ischemia in the transverse colon.</li>
		<li>Left posterior of SMA approach: For this approach, follow steps 2.3.7-2.3.18.</li>
		<li>Place the entire small bowel on the left side to facilitate exposure and the dissection of the distal duodenum proximal to the ligament of Treitz.</li>
		<li>Divide the proximal jejunum with a stapler.</li>
		<li>Expose the SMA by tracing along the jejunal artery.</li>
		<li>Place a Fr8 catheter for hanging for encircling the dorsal aspect of the SMA and SMV.</li>
		<li>Pull the catheter to the upper right side to allow SMA dissection on its periadventitial plane on the anterior-left margin and its separation from the mesopancreas.</li>
		<li>Ligate the first jejunal artery (FJA) involved by the tumor and sacrifice.</li>
		<li>Identify the first jejunal vein (FJV) and the inferior pancreaticoduodenal veins (IPDV), which are the branches from the dorsal side of the SMV.</li>
		<li>Ligate and dissect the IPDV.</li>
		<li>Circumferentially dissect the SMA to identify the inferior pancreaticoduodenal artery (IPDA), which either forms a common trunk with the FJA or arises directly from the SMA (Figure 3).</li>
		<li>Sacrifice the IPDA.</li>
		<li>At the left posterior approach, dissect the SMA, which is identified at its origin above the LRV, free from the mesopancreas.</li>
		<li>In cases that the tumor invades the mesocolon, hang the SMV and SMA through a rubber band followed by coring out of the mesocolon.</li>
		<li>Supracolic median-anterior of SMA approach: For this approach, follow steps 2.3.20-2.3.34.</li>
		<li>Start the SMA approach from the upper colon region.</li>
		<li>Explore the gap between the pancreatic neck and SMV at the lower edge of the pancreas.</li>
		<li>Use a stapler device to divide the stomach 3-5 cm away from the pylorus.</li>
		<li>Remove the gallbladder.</li>
		<li>Divide the common bile duct (CBD) as conventional PD. Take a frozen pathologic examination of the bile duct stump. Apply the laparoscopic bulldog clamps to occlude the CBD temporarily.</li>
		<li>Dissect the hepatoduodenal ligament. Perform a lymphadenectomy along the CHA, the proper hepatic artery (PHA), and the PV.</li>
		<li>Ligate and dissect the right gastric artery.</li>
		<li>Identify the gastroduodenal artery (GDA) at the point where the PHA branches from the CHA.</li>
		<li>Doubly ligate or suture the GDA to minimize the chance of subsequent erosion and bleeding.</li>
		<li>Transect the neck of the pancreas with an ultrasound knife. Take a frozen pathologic examination of the pancreatic stump.</li>
		<li>Suspend the splenic vein (PV) using a rubber band.</li>
		<li>Set up a diamond-shaped window by retracting the transverse mesocolon caudally, the PV cranially, the SMV rightward, and the SMA leftward.</li>
		<li>During the supracolic median-anterior of the SMA approach, dissect the right and dorsal aspects of the SMA within this diamond-shaped window.</li>
		<li>Resect the fat and fibrous tissues around SMA and CA from the caudal side to the cephalic side.</li>
		<li>Ligate and transect the uncinate process artery (UPA) under the PV, leaving the specimen attached to the PV/SMV.</li>
		<li>Combining with median-anterior and left-posterior approaches to the SMA, expose the origin of the IPDA or the common trunk of the IPDA and FJA easily. 
		NOTE: During this procedure, the GDA, UPA, and IPDA have been completely ligated. Furthermore, all the tumor artery inflows are completely occluded till this moment to reduce bleeding.</li>
		<li>Use laparoscopic bulldog clamps to temporarily clamp the PV, the splenic vein, and the SMV. Transect the involved vein with sufficient margin.</li>
		<li>Afterward, cut off the lymphatics and dissect the duodenum from the retroperitoneum.</li>
		<li>Finally, resect the tumor in situ and remove en bloc following the oncologic principles of No-Touch2,6,7.</li>
	</ol>
	</li>
	<li>Reconstruction phase
	<ol>
		<li>Measure the length of the venous defect with a soft ruler. If the defect exceeds 3 cm, consider using an artificial interposition graft.</li>
		<li>Perform SMV reconstruction from caudal to cephalic by continuous suture using 5-0 prolene sutures (Figure 4).</li>
		<li>Reconstruct the digestive tract by Child&#39;s method9.</li>
		<li>An end-to-side, perform single-layer running suturing hepaticojejunostomy with 4-0 absorbable sutures.</li>
		<li>Conduct pancreatic anastomosis by duct-to-mucosal, end-to-side pancreaticojejunostomy within an internal stent9.</li>
		<li>After a side-to-side gastrojejunal anastomosis, place three drainages near the anastomosis.</li>
	</ol>
	</li>
	<li>Bag the specimen and retrieve it through the Pfannenstiel incision.</li>
</ol>

<ol>
	<li>Hirota, M., 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=Pancreatectomy+using+the+no-touch+isolation+technique+followed+by+extensive+intraoperative+peritoneal+lavage+to+prevent+cancer+cell+dissemination:+a+pilot+study.">Pancreatectomy using the no-touch isolation technique followed by extensive intraoperative peritoneal lavage to prevent cancer cell dissemination: a pilot study.</a> Journal of the Pancreas. 6 (2), 143-151 (2005).</li><li>Gall, T. M., 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=Reduced+dissemination+of+circulating+tumor+cells+with+no-touch+isolation+surgical+technique+in+patients+with+pancreatic+cancer.">Reduced dissemination of circulating tumor cells with no-touch isolation surgical technique in patients with pancreatic cancer.</a> JAMA Surgery. 149 (5), 482-485 (2014).</li><li>Soeth, E., 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=Detection+of+tumor+cell+dissemination+in+pancreatic+ductal+carcinoma+patients+by+CK+20+RT-PCR+indicates+poor+survival.">Detection of tumor cell dissemination in pancreatic ductal carcinoma patients by CK 20 RT-PCR indicates poor survival.</a> Journal of Cancer Research and Clinical Oncology. 131 (10), 669-676 (2005).</li><li>Yap, T. A., Lorente, D., Omlin, A., Olmos, D., de Bono, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+tumor+cells:+a+multifunctional+biomarker.">Circulating tumor cells: a multifunctional biomarker.</a> Clinical Cancer Research. 20 (10), 2553-2568 (2014).</li><li>Fujita, 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=Laparoscopic+right+hemicolectomy+with+radical+lymph+node+dissection+using+the+no-touch+isolation+technique+for+advanced+colon+cancer.">Laparoscopic right hemicolectomy with radical lymph node dissection using the no-touch isolation technique for advanced colon cancer.</a> Surgery Today. 31 (1), 93-96 (2001).</li><li>Hirota, M., 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=Pancreatoduodenectomy+using+a+no-touch+isolation+technique.">Pancreatoduodenectomy using a no-touch isolation technique.</a> The American Journal of Surgery. 199 (5), 65-68 (2010).</li><li>Kuroki, T., Eguchi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No-touch+isolation+techniques+for+pancreatic+cancer.">No-touch isolation techniques for pancreatic cancer.</a> Surgery Today. 47 (1), 8-13 (2017).</li><li>Tan, Z. 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=Clinical+experience+of+laparoscopic+pancreatoduodenectomy+via+orthotopic+resection.">Clinical experience of laparoscopic pancreatoduodenectomy via orthotopic resection.</a> Zhonghua Wai Ke Za Zhi [Chinese Journal of Surgery. 58 (10), 782-786 (2020).</li><li>Osada, 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=Reconstruction+method+after+pancreaticoduodenectomy.+Idea+to+prevent+serious+complications.">Reconstruction method after pancreaticoduodenectomy. Idea to prevent serious complications.</a> Journal of the pancreas. 13 (1), 1-6 (2012).</li><li>Mizrahi, J. D., Surana, R., Valle, J. W., Shroff, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pancreatic+cancer.">Pancreatic cancer.</a> Lancet. 395 (10242), 2008-2020 (2020).</li><li>Tempero, M. 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=Pancreatic+adenocarcinoma,+version+2.2021,+NCCN+clinical+practice+guidelines+in+oncology.">Pancreatic adenocarcinoma, version 2.2021, NCCN clinical practice guidelines in oncology.</a> Journal of the National Comprehensive Cancer Network JNCCN. 19 (4), 439-457 (2021).</li><li>Bockhorn, M., 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=Borderline+resectable+pancreatic+cancer:+a+consensus+statement+by+the+International+Study+Group+of+Pancreatic+Surgery+(ISGPS).">Borderline resectable pancreatic cancer: a consensus statement by the International Study Group of Pancreatic Surgery (ISGPS).</a> Surgery. 155 (6), 977-988 (2014).</li><li>Tee, M. C., 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=Laparoscopic+pancreatoduodenectomy+does+not+completely+mitigate+increased+perioperative+risks+in+elderly+patients.">Laparoscopic pancreatoduodenectomy does not completely mitigate increased perioperative risks in elderly patients.</a> HPB (Oxford). The Official Journal of the International Hepato Pancreato Biliary Association. 17 (10), 909-918 (2015).</li><li>Palanivelu, C., 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=Randomized+clinical+trial+of+laparoscopic+versus+open+pancreatoduodenectomy+for+periampullary+tumours.">Randomized clinical trial of laparoscopic versus open pancreatoduodenectomy for periampullary tumours.</a> The British Journal of Surgery. 104 (11), 1443-1450 (2017).</li><li>Wang, M., 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=Laparoscopic+versus+open+pancreatoduodenectomy+for+pancreatic+or+periampullary+tumours:+A+multicentre,+open-label,+randomised+controlled+trial.">Laparoscopic versus open pancreatoduodenectomy for pancreatic or periampullary tumours: A multicentre, open-label, randomised controlled trial.</a> The Lancet. Gastroenterology &amp; Hepatology. 6 (6), 438-447 (2021).</li></ol>

The authors have nothing to disclose.

PDAC is one of the most lethal malignant diseases. Despite the fact that the overall 5-year survival rates are still unsatisfactory, surgery remains the only curative therapeutic method till now10. According to the National Comprehensive Cancer Network (NCCN) and International Study Group of Pancreatic Cancer (ISGPS), patients diagnosed with PDAC should be defined as borderline resectable cases while the portal-superior mesenteric vein is suspiciously involved, and in order to improve R0 resection...

Pancreaticoduodenectomy (PD) is a standard surgical procedure for cancer in the pancreaticoduodenal region. The Kocher maneuver is widely used for the efficient exposure of the duodenum and pancreatic head during conventional PD. The mobilization and squeezing of the pancreaticoduodenal area during surgery may cause metastasis of the tumor cells before the ligation of surrounding vessels1. A recent study had shown that the tumor cells had the potential possibility of being squeezed into the portal vein (PV) because of the handling and squeezing of the tumor area by the surgeons, which might further increase the risk of liver metastasis after surgery2.
With the development of biomedical technology, a scientist could detect the spread of solid tumor cells, including pancreatic cancer cells, into the vessels as circulating tumor cells (CTC)3,4.
No-Touch isolation procedures, which have been used in colon cancer, might prevent the dissemination of cancer cells, such as circulating tumor cells, from the primary tumor5. Several studies have reported the use of a no-touch isolation technique for pancreatic head cancer during laparotomy pancreaticoduodenectomy6,7. The concept of this procedure is that the surgeon does not touch the duodenum and pancreatic head region (including the tumor) before ligating and dissecting the vessels (arteries and veins) around the pancreatic head.
No-Touch isolation techniques have been reported in LPD for pancreaticoduodenal region neoplasm8. We herein present a modified in-situ No-Touch isolation LPD with partial resection and reconstruction of SMV for pancreatic cancer after neoadjuvant therapy, which dissects all the inflow arteries first, transects the involved vein with sufficient margin, resects the tumor in-situ, and removes the specimen en-bloc.
The goal and advantages of this method are to ensure that all steps follow the oncologic principles of No-Touch in order to decrease the risk of metastasis of the tumor cells. The rationale behind the development and use of this technique is that the tumor should be mobilized at the final stage, including resecting the tumor in situ and removing the specimen en bloc after tumor inflow arteries and outflow veins are occluded. However, as this procedure requires complex resection and reconstruction techniques when surgeons decide whether to use this method, they need to estimate their own situations such as the learning curve, tumor type, vascular condition, and other factors.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D Laparoscope</td>
			<td>STORZ</td>
			<td>TC200,TC302</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable hemostat</td>
			<td>ETHICON, LLC</td>
			<td>2 in x 4 in</td>
			<td></td>
		</tr>
		<tr>
			<td>Artificial Interposition Graft</td>
			<td>W.L.Gore &#38; Associates, Inc.</td>
			<td>IRTH084040W</td>
			<td></td>
		</tr>
		<tr>
			<td>Drainage tube</td>
			<td>Aiyuan</td>
			<td>424280</td>
			<td></td>
		</tr>
		<tr>
			<td>Echelon Flex Powered Plus Articulating Endoscopic Linear Cutter and Endopath Echelon Endoscopic Linear Cutter Reloads with Gripping Surface Technology</td>
			<td>Ethicon Endo-Surgery</td>
			<td>ECR60G/GST60G</td>
			<td></td>
		</tr>
		<tr>
			<td>Energy Platform</td>
			<td>COVIDIEN ForceTriad Energy Platform</td>
			<td>T2131469EX</td>
			<td></td>
		</tr>
		<tr>
			<td>HARMONIC ACE Ultrasonic Surgical Devices</td>
			<td>Ethicon Endo-Surgery</td>
			<td>HAR36</td>
			<td></td>
		</tr>
		<tr>
			<td>Laparoscopic forceps</td>
			<td>Gimmi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Laparoscopic right angle forceps</td>
			<td>KARL STORZ</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Laparoscopic scissors</td>
			<td>AESCULAP</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Latex T-shape Catheter</td>
			<td>ZHANJIANG STAR ENTERPRISE CO., LTD.</td>
			<td>24Fr</td>
			<td></td>
		</tr>
		<tr>
			<td>Ligating Clips</td>
			<td>Teleflex Medical</td>
			<td>5,44,22,05,44,23,05,44,000</td>
			<td></td>
		</tr>
		<tr>
			<td>PDSII</td>
			<td>Ethicon, LLC</td>
			<td>W9109</td>
			<td></td>
		</tr>
		<tr>
			<td>PROLENE</td>
			<td>Ethicon, LLC</td>
			<td>W8556</td>
			<td></td>
		</tr>
		<tr>
			<td>Trocar</td>
			<td>Surgaid</td>
			<td>NPCS-100-1-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic Surgical &#38; Electrosurgical Generator</td>
			<td>Ethicon Endo-Surgery</td>
			<td>GEN11CN</td>
			<td></td>
		</tr>
	</tbody>
</table>

laparoscopic pancreatoduodenectomy for pancreatic cancer using in-situ no-touch isolation technique

Laparoscopic pancreatoduodenectomy (LPD) is a standard radical operation for pancreatic head malignant tumors by now. Due to the complex laparoscopic resection and reconstruction techniques, it is difficult to perform LPD for patients with locally advanced pancreatic head cancer after neoadjuvant therapy. Our team initiates LPD using the in-situ No-Touch isolation technique. The innovation and optimization of this modified No-Touch isolation technique emphasize exploring the distal section of superior mesenteric vein&#160;(SMV) and the left side of the superior mesenteric artery (SMA) prior to evaluating the resectability by subcolonic mesenteric approach, which is an ideal exploring approach. After that, we use the median-anterior, and left-posterior of SMA approaches to cut off the blood flow of the pancreatic head to make the tumor isolated intact, then move and dissect the tumor. It is a process fitting the surgical principle of tumor-free. This article aims to demonstrate the feasibility and safety of performing LPD using the in-situ No-Touch isolation technique, which might elevate the R0 resection rate. It is an oncological ideal operation process.

A 55-year-old man with upper abdominal pain and marasmus was diagnosed with a 4.2 cm x 3.1 cm tumor in the uncinate process of the pancreas, and the SMV was involved over 180&#176; (Figure 5). The patient was previously healthy and had a relatively normal body mass index (19.47 kg/m2). No distant metastasis was found on the preoperative contrast-enhanced CT scan. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) was performed to acquire the pathology diagnosis of adeno...

Watch this Scientific Journal Video about Laparoscopic Pancreatoduodenectomy for Pancreatic Cancer Using In-Situ No-Touch Isolation Technique at JoVE.com

Laparoscopic Pancreatoduodenectomy for Pancreatic Cancer Using In-Situ No-Touch Isolation Technique

No-Touch isolation procedures might prevent the dissemination of cancer cells from the primary tumor. However, these techniques are not widely accepted in laparoscopic pancreatoduodenectomy (LPD) by now. We herein present in-situ No-Touch isolation LPD with partial resection and reconstruction of the superior mesenteric vein (SMV) for pancreatic cancer after neoadjuvant therapy.

Laparoscopic Pancreatoduodenectomy for Pancreatic Cancer Using In-Situ No-Touch Isolation Technique

Laparoscopic pancreatoduodenectomy (LPD) is a standard radical operation for pancreatic head malignant tumors by now. Due to the complex laparoscopic ...

laparoscopic-pancreatoduodenectomy-for-pancreatic-cancer-using-situ

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1.8K Views.  The Second Affiliated Hospital of Guangzhou University of Chinese Medicine. No-Touch isolation procedures might prevent the dissemination of cancer cells from the primary tumor. However, these techniques are not widely accepted in laparoscopic pancreatoduodenectomy (LPD) by now. We herein present in-situ No-Touch isolation LPD with partial resection and reconstruction of the superior mesenteric vein (SMV) for pancreatic cancer after neoadjuvant therapy.

Video: Laparoscopic Pancreatoduodenectomy for Pancreatic Cancer Using In-Situ No-Touch Isolation Technique

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Clinicopathological Analysis of miRNA Expression in Breast Cancer Tissues by Using miRNA In Situ Hybridization

In this article, we describe a detailed protocol for miRNA detection in breast cancer tissue using in situ hybridization with a digoxigenin-labelled LNA (Locked Nucleic Acid) probe. The probe was recognized by anti-DIG alkaline phosphatase antibodies and later developed using alkaline phosphatase substrate producing fluorescence signals. Here we utilized miRNA in situ hybridization (MISH) technique to analyze expression of miR-489 in tissues from breast cancer patients. This technique can detect the localization of miRNA of interest in individual tissue samples. This technique can be used to compare the expression of desired miRNA in tumor tissue with that in adjacent normal tissue and to identify the specific structures responsible for expressing this miRNA. This technique can be very useful in answering certain clinical questions, such as role of specific miRNA in the development of cancer. Our results indicate that mammary epithelial cells express significantly higher levels of miR-489 than adjacent tumor cells.

Clinicopathological Analysis of miRNA Expression in Breast Cancer Tissues by Using miRNA In Situ Hybridization

Here we present a protocol to detect miRNA expression in breast cancer patient samples using miRNA in situ hybridization.

In this article, we describe a detailed protocol for miRNA detection in breast cancer tissue using in situ hybridization with a digoxigenin-labelled LNA ...

Laparoscopic Technique for Serial Collection of Liver and Mesenteric Lymph Nodes in Macaques

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them immunologically unique. The GI tract and associated MLN are sites of early viral replication in human immunodeficiency virus (HIV) infection and the MLN are likely important reservoir sites that harbor latently-infected cells even after prolonged antiretroviral therapy (ART). The liver has been shown to play a significant role in immune responses to lentiviruses and appears to play a significant role in clearance of virus from circulation. Nonhuman primate (NHP) models for HIV and Acquired Immunodeficiency Syndrome (AIDS) closely mimic these aspects of HIV infection and serial longitudinal sampling of primary sites of viral replication and the associated immune responses in this model will help to elucidate critical events in infection, pathogenesis, and the impact of various intervention strategies on these events. Current published techniques to sample liver and MLN together involve major surgery and/or necropsy, which limits the ability to investigate these important sites in a serial fashion in the same animal. We have previously described a laparoscopic technique for collection of MLN. Here, we describe a minimally invasive laparoscopic technique for serial longitudinal sampling of liver and MLN through the same two port locations required for the collection of MLN. The use of the same two ports minimizes the impact to the animals as no additional incisions are required. This technique can be used with increased sampling frequency compared to major abdominal surgery and reduces the potential for surgical complications and associated local and systemic inflammatory responses that could complicate interpretation of results. This procedure has potential to facilitate studies involving NHP models while improving animal welfare.

Here, we describe a minimally invasive laparoscopic technique for serial sampling of liver and mesenteric lymph nodes (MLN) in macaques that allows for increased sampling frequency, and reduces the potential for surgical complications when compared to performing a laparotomy.

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them ...

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.

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

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

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

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

Autologous arteriovenous fistula (AVF) is the primary and best option to obtain vascular access for hemodialysis treatment; other options are ...

Laparoscopic Radical Antegrade Modular Pancreatosplenectomy via Dorsal-Caudal Artery Approach for Pancreatic Neck-Body Cancer

Laparoscopic radical resection of the pancreatic neck is one of the most complicated radical operations for pancreatic cancer, especially for patients who have had neoadjuvant chemotherapy. Here, we present a technique to perform laparoscopic radical antegrade modular pancreatosplenectomy (L-RAMPS) using the dorsal-caudal artery approach by making full use of the high-definition vision and operation modes of the laparoscope.
The innovation and optimization of this operation are provided in the protocol. Priority should be given to the dorsal resection plane, including the dorsal side of the superior mesenteric artery (SMA), the dorsal side of the pancreatic head, the root of the celiac artery (CeA), the ventral side of the left renal vessels, and the renal hilum. On the condition that the operation for pancreatic neck-body cancer is feasible and safe, the second step is to perform tumor resection en bloc surrounding the SMA and CeA from the caudal to the cephalic side to increase the rate of R0 (radical zero) resection and further prognosis.

Laparoscopic Radical Antegrade Modular Pancreatosplenectomy via Dorsal-Caudal Artery Approach for Pancreatic Neck-Body Cancer

With advancements in laparoscopic techniques, laparoscopic radical antegrade modular pancreatosplenectomy (L-RAMPS) has been widely recognized. However, owing to several technical difficulties in this procedure, the artery-first approach in L-RAMPS still remains uncommon. Here, we developed the dorsal-caudal artery approach for L-RAMPS, which might be safe and beneficial for pancreatic neck tumors.

Laparoscopic radical resection of the pancreatic neck is one of the most complicated radical operations for pancreatic cancer, especially for patients who ...

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.

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

Application of Laparoscopic Hepatectomy Combined with Intraoperative Microwave Ablation in Colorectal Cancer Liver Metastasis

Laparoscopic hepatectomy is a common treatment for colorectal cancer liver metastasis. Previously, a sufficient number of functional liver masses had to be maintained during laparoscopic hepatectomy, with a residual liver volume of &gt;40% in cirrhotic patients and &gt;30% in non-cirrhotic patients. The high incidence of complications such as bleeding, bile leakage, or liver failure due to the exposure and difficulty of the resection of specific liver segments such as S2 and S7 reduces the success rate of liver resection. At present, microwave ablation is mainly applied in the treatment of liver metastasis using a percutaneous approach, which makes it difficult to identify hidden parts or small lesions. For some liver segments, the percutaneous puncture of liver segment 7 (S7) is likely to pass through the thoracic cavity, and the percutaneous puncture of liver segment 2 (S2) adjacent to the diaphragm is likely to injure the diaphragm and heart; these issues restrict the application of percutaneous ablation in colorectal cancer liver metastasis. Considering multiple lesions, laparoscopic microwave ablation combined with hepatectomy was performed in this study. The location of the lesions was determined by contrast-enhanced ultrasound under laparoscopy, and small lesions that were difficult to detect before the operation were identified. For the scattered lesions, which had diameters less than 3 cm and were difficult to resect, ablation was adopted to substitute hepatectomy. This technique helped to more explicitly locate the tumors, simplified the operation procedures, reduced the risk of complications such as bleeding and bile leakage, shortened the operation time, accelerated the postoperative recovery, significantly improved the success rate of operation, and enhanced the clinical prognosis of colorectal cancer liver metastasis by surgical resection.

This protocol describes the laparoscopic resection of colorectal cancer liver metastases combined with ultrasound-guided microwave ablation. This technique can safely, effectively, and accurately treat refractory liver metastases &lt;3 cm, reduce postoperative complications, and accelerate the postoperative rehabilitation of patients.

Laparoscopic hepatectomy is a common treatment for colorectal cancer liver metastasis. Previously, a sufficient number of functional liver masses had to ...

Management of the Uncinate Process in No-Touch Laparoscopic Pancreaticoduodenectomy

Laparoscopic pancreatoduodenectomy (LPD) is a demanding abdominal operation that necessitates meticulous surgical skills and teamwork. The management of the pancreatic uncinate process is one of the most important and difficult processes in LPD because of its deep anatomical location and difficult exposure. Complete resection of the uncinate process and mesopancreas has become the cornerstone of LPD. In particular, it is even more difficult to avoid positive surgical margins and incomplete lymph node dissection when the tumor is located in the uncinate process. No-touch LPD, which is an ideal oncological operation process fitting the "tumor-free" principle, has been reported by our group previously. This article introduces the management of the uncinate process in no-touch LPD. Based on the multi-angle arterial approach, in this protocol, the median-anterior and left-posterior approaches to the SMA are used to correctly deal with the important vascular structure, the inferior pancreaticoduodenal artery (IPDA), in order to ensure the safe and complete excision of the uncinate process and mesopancreas. For the achievement of the no-touch isolation technique in LPD, the pancreatic head and the blood supply to the duodenal region must be severed at the very early stage of the operation; after that, the tumor can be isolated intact, resection can be performed in situ, and finally, the tissue can be removed en bloc. This paper aims to show the distinctive ways to manage the uncinate process in no-touch LPD and investigate the viability and safety of this approach. Moreover, the technique may increase the R0 resection rate.

Complete resection of the uncinate process and mesopancreas is one of the most important and difficult processes in laparoscopic pancreatoduodenectomy (LPD). This article presents a method for managing the uncinate process in no-touch LPD using the median-anterior and left-posterior approaches to the superior mesenteric artery (SMA).

Laparoscopic pancreatoduodenectomy (LPD) is a demanding abdominal operation that necessitates meticulous surgical skills and teamwork. The management of ...