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Microsurgical Techniques and Immunological Principles
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Design the rings according to the parameters of rat SHVC with a major axis diameter, minor axis diameter, thickness and weight of 8 mm, 5. Ensure that the cuff body is 5 mm long with a 2 mm cuff extension. The Stent Figure 2 Prepare the stent for anastomosis of the BD by cutting G intravenous cannula with a length of 5 mm to produce slant at both ends.
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Experimental Animals Use male Sprague Dawley rats weighing between and g as donors and recipients. Keep the rats in climate-controlled rooms with free access to food and water before surgery. Donor Operation Inject buprenorphine 0. Anesthetize the donor rat by isoflurane inhalation.
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Confirm depth of anesthesia by performing a toe pinch and a skin pinch. Shave the entire abdominal skin of the rat using an electric shaver to allow for cleaner exposure. Disinfect the corresponding skin with a povidone-iodine solution. Open the abdominal cavity via a cruciform incision using surgical scissors from the root of the penis extending to 1 cm above the xiphisternum along linea alba , extending from the midpoint of the longitudinal incision to the midaxillary line.
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Exteriorize the gastrointestinal tract to the left and cover it with wet gauze. Clamp and pull the xiphisternum toward the head using a hemostatic forceps. Dissect the falciform ligament and the connective tissues around the liver. Isolate and ligate the left inferior phrenic vein with a silk suture. Insert the stent into the bile duct. Clamp the BD just above the point where the gastroduodenal vein joins the portal vein and pull it to maintain a little tension using a microforceps.
Then perform a "V" incision of 1 mm length on the anterior wall about 5 mm proximal to the biliary confluence by a micro scissors. Insert the stent into the lumen of the bile duct using a curved microforceps.
An easy and safe model of kidney transplantation in rats
Make sure at least a half of the stent lies outside the bile duct. Secure the stent with a silk suture. Cut one end of the suture. Keep the other one to hold during later anastomosis. Transect the bile duct below the stent and make sure the bile can come out of the stent. Isolate the IHVC down to the left renal vein level.
Separate and ligate the right suprarenal vein and the right renal vein with a silk suture. Heparinize the rat by injecting 50 U of heparin diluted in 2 mL of normal saline solution through the dorsal vein of the penis. Perfuse the liver.
Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique
Isolate the abdominal aorta below the left renal vein. Insert a G catheter into the aorta. Perform a thoracotomy, clamp the thoracic aorta using a microforceps. Dissect the connective tissue surrounding the PV by two micro forceps. Ligate and divide the pyloric veins with an prolene suture. Transect the PV at the level of the splenic vein. Excise the donor liver and rinse it with cold normal saline solution. Insert the IHVC into its cuff with the cuff extension pointing towards the liver.
Position the cuff extension on the posterior wall of IHVC. Fix the cuff extension and the IHVC with a curved microforceps itself attached using a file clip to the bath container. Evert the distal end of the IHVC over the cuff body using two micro forceps. Secure the edge with a silk suture. Make sure the IHVC is not twisted during the process. Repeat step 3. Pull the diaphragm through the four forceps and trim the SHVC. Pass the SHVC through the magnetic ring using the titanium-alloy micro forceps. Evert the distal end of the SHVC over the ring.
Secure the ring with a silk suture. Make sure the SHVC is not twisted during the process.
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Remove the excess diaphragm around the liver. Inject buprenorphine 0. Shave the entire abdominal skin of the rat. Open the abdominal cavity via a 4 cm midline incision. Place the abdominal retractors. Pull the ribs as far apart as possible from the midline.
Dissect the connective tissues and ligaments around the liver. Ligate and divide the hepato-esophageal ligament. Ligate and divide the hepatic artery with an prolene suture. Isolate the common bile duct from the first hepatic portal. Place a silk suture around the common bile duct just below its division and ligate the common bile duct. Dissect the IHVC down to the right renal vein. Ligate and divide the right adrenal vein.
Place a strap in the SHVC of the liver. Clamp the IHVC just above the right renal vein with microvessel clips. Clamp the PV at the level of the pyloric vein with microvessel clips. To flush the blood out of the liver, slowly inject 2 mL of normal saline through the PV. Drag down the liver by pulling the previously placed strap.
Excise the liver by transecting the SHVC just above the liver.
Remove the recipient liver quickly. Attach a magnetic ring to the recipient's SHVC. Insert the recipient's remaining SHVC into a magnetic ring. Fix the magnetic ring on a Satinsky clamp by magnetic attraction. Evert the SHVC over the magnetic ring until the vessel covers the edge of the ring.
Place the donor liver graft orthotopically and cover it with a cold wet gauze. Fill the lumen of the donor's and the recipient's SHVCs with normal saline solution. Remove air bubbles thoroughly to prevent air embolism. Reconstruct the PV.