Role of ubiquitin-conjugating enzyme E2T in the carcinogenesis and progression of pancreatic cancer.

Ubiquitin-conjugating enzyme E2T (UBE2T) is a recently discovered oncogenic protein. Numerous studies reported that UBE2T is highly expressed in various types of human cancer; however, its role in the carcinogenesis and progression of pancreatic cancer remains unknown. The aim of the present study was to investigate the role of UBE2T in pancreatic cancer progression through in vitro experiments in pancreatic cancer tissues and cell lines. The results obtained in the present study demonstrated that UBE2T served an important role in the initiation and progression of pancreatic cancer. Furthermore, increased expression of UBE2T in human pancreatic cancer tissues and pancreatic cancer cells was observed compared with normal tissues and cells. The effect of upregulating and downregulating UBE2T in pancreatic cancer cell lines was investigated using the MTT, wound-healing and migration and invasion assays. The results demonstrated that overexpression of UBE2T significantly promoted pancreatic cancer cell proliferation, migration and invasion compared with controls. However, UBE2T downregulation resulted in the inhibition of pancreatic cancer cell proliferation, migration and invasion. In addition, the results demonstrated that UBE2T may promote the epithelial mesenchymal transition in pancreatic cancer cells.

Hepatocyte isolation from resected benign tissues: Results of a 5-year experience.

AIM: To analyze retrospectively a 5-year experience of human hepatocyte isolation from resected liver tissues with benign disease. METHODS: We established a method of modified four-step retrograde perfusion to isolate primary human hepatocytes. Samples were collected from the resected livers of patients with intrahepatic duct calculi (n = 7) and liver hemangioma (n = 17). Only the samples weighing ≥ 15 g were considered suitable for hepatocyte isolation. By using the standard trypan blue exclusion technique, hepatocyte viability and yield were immediately determined after isolation. RESULTS: Twenty-four liver specimens, weighing 15-42 g, were immediately taken from the margin of the removed samples and transferred to the laboratory for hepatocyte isolation. Warm ischemia time was 5-35 min and cold ischemia time was 15-45 min. For the 7 samples of intrahepatic duct calculi, the method resulted in a hepatocyte yield of 3.49 ± 2.31 × 10(6) hepatocytes/g liver, with 76.4% ± 10.7% viability. The 17 samples of liver hemangioma had significantly higher yield of cells (5.4 ± 1.71 × 10(6) cells/g vs 3.49 ± 2.31 × 10(6) cells/g, P < 0.05) than the samples of intrahepatic duct calculi. However, there seems to be no clear difference in cell viability (80.3% ± 9.67% vs 76.4% ± 10.7%, P > 0.05). We obtained a cell yield of 5.31 ± 1.87 × 10(6) hepatocytes/g liver when the samples weighed > 20 g. However, for the tissues weighing ≤ 20 g, a reduction in yield was found (3.08 ± 1.86 × 10(6) cells/g vs 5.31 ± 1.87 × 10(6) cells/g, P < 0.05). CONCLUSION: Benign diseased livers are valuable sources for large-number hepatocyte isolation. Our study represents the largest number of primary human hepatocytes isolated from resected specimens from patients with benign liver disease. We evaluated the effect of donor liver characteristics on cell isolation, and we found that samples of liver hemangioma can provide better results than intrahepatic duct calculi, in terms of cell yield. Furthermore, the size of the tissues can affect the outcome of hepatocyte isolation.

Reversible immortalization of human hepatocytes mediated by retroviral transfer and site-specific recombination.

AIM: To establish a method for the reversible immortalization of human hepatocytes, which may offer a good and safe source of hepatocytes for practical applications. METHODS: We successfully isolated primary human hepatocytes from surgically resected liver tissue taken from a patient with liver hemangiomas. The freshly isolated cells were then immortalized with retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40T) and hygromycin-resistance genes flanked by paired loxP recombination targets. RESULTS: The freshly isolated hepatocytes with high viability (85%) were successfully immortalized using retroviral gene transfer of SV40T. SV40T in the immortalized cells was then excised by Cre/loxP site-specific recombination. This cell population exhibited the characteristics of differentiated hepatocytes. CONCLUSION: We successfully established reversibly immortalized human hepatocytes, which will provide an unlimited supply of cells for practical applications.

[Thumb replantation through bridging ulnar proper digital artery and dorsal carpal branch of radial artery].

OBJECTIVE: To introduce treatment method of replantation of severed thumb trauma with proximal arterial, and to evaluate its efficacy. METHODS: From February 2007 to March 2009,13 patients with severed thumb in serious injury of proximal arterial were treated with vein graft of volar forearm,bridging between dorsal carpal branch of radial artery on nasopharyngeal fossa and distal stump of ulnar proper digital artery. Among them, there were 11 males and 2 females with an average age of 34.5 years ranging from 16 to 50 years. Seven cases were in the left thumb,6 in the right thumb. Eight cases were complete separation, incomplete separation in 5 cases. Ten cases recieved emergency reimplantation, 3 cases with the arterial crisis after conventional replantation were explorated and repaired. RESULTS: Thumb of 13 cases all survived. All patients were followed-up for 8 to 17 months (averaged, 11 months). Replantation thumb obtained satisfactory appearance. According to Chinese Medical Association Society of Hand Surgery Trial criteria, the results were excellent in 9 fingers, good in 3 fingers, 1 poor finger. Two point discrimination of finger pulp was 5 to 8 mm (averaged 6.5 mm). CONCLUSION: By repairing artery of thumb with vein graft of volar forearm, bridging between dorsal carpal branch of radial artery on nasopharyngeal fossa and distal stump of ulnar proper digital artery,complex severed thumb replants on surgery position comfortable,without affecting the blood supply of the hand, expanding the indications for replantation and improving the success rate of replantation.

[Treatment of carpal tunnel syndrome with mini-incision decompression].

OBJECTIVE: To introduce the method of the transverse carpal ligament (TCL) transection through palmar mini-incision for carpal tunnel syndrome (CTS) and evaluate its efficacy and safety. METHODS: From January 2006 to September 2007, 15 patients (18 hands) with CTS were treated with palmar longitudinal mini-incision decompression. There were 2 males and 13 females, with an average age of 48 years (ranged 34 to 69) and an average course of 18 months (ranged 8 to 26). The main clinical symptom including the radial numbness or pain of the 3 and a half fingers, wrist pain and radiation to the forearm, the night awake history of numbness, thenar muscle atrophy, positive Tinel sign and Phalen sign. Median nerve electrophysiology showed that sensory nerve conduction velocity (SCV) slowed down and sensory nerve action potential (SNAP) decreased or missed, short abductor muscle of thumb had spontaneous potential in severe cases. Clinical effect were evaluated according to the Global symptom score (GSS) scoring in aspect of pain, numbness, paraesthesia, weakness and nocturnal awakening. RESULTS: Postoperative wound smoothly healed in all patients and no complications occurred. All patients were followed up from 20 to 28 months with an average of 24 months. Symptoms of 1 patient had not incomplete relief, other patient's symptoms disappeared and muscle force of abductor pollicis brevis reinforced. Postoperative GSS scoring obviously improved than preoperative (P < 0.05). CONCLUSION: Treatment of carpal tunnel syndrome through palmar mini-incision decompression has advantages such as higher safe, shorter operative time, less invasive, smaller scar, which can cut off carpi transversum ligament and thoroughly decompress median nerve under direct sight, it is a safe and effective operative approach.

Extended local resection for treatment of periampullary carcinoma of vater.

BACKGROUND: The trauma caused by pancreatoduodenectomy for periampullary carcinoma of vater is often severe and extensive. The purpose of this study was to evaluate the effect of extended local resection in the treatment of periampullary carcinoma of vater. METHODS: The extra-hepaticobiliary tract, the confluence of the pancreatic and biliary duct, vater ampulla and duodenal papilla were resected en bloc in 8 patients with periampullary carcinoma from 1995 to 1998. RESULTS: One patient died perioperatively. Duodenal obstruction developed postoperatively in one of 7 survived patients and was relieved after reoperation. All the 7 patients were followed up for more than 6 months without recurrence. CONCLUSION: Extended local resection fulfils the task of radical treatment of periampullary malignancy.

Surgical treatment for uncinate process carcinoma of the pancreas.

OBJECTIVES: To analyze the clinical features of uncinate process carcinoma of the pancreas and to improve the resection rate. METHODS: From January 1990 to June 1999, 10 patients with pancreas uncinate process carcinoma received Whipple's operation. Portal vein (PV) resection and reanastomosis were performed in 5 patients, and the resected length varied from 2.0 to 4.2 cm. Two patients underwent PV lateral wall partial resection. RESULTS: Among the 7 patients undergoing PV resection, 1 died of hepatic failure 3 days after operation. One patient suffered from postoperative chylous ascites. These 6 patients survived 13 to 29 months postoperatively. Among the 3 patients without PV resection, 2 survived 13 months and 14 months respectively. One patient was alive by the end of follow-up for 11.5 months postoperatively. CONCLUSION: Although uncinate process carcinoma of the pancreas has a tendency to invade the adjacent PV and superior mesentery vein, it should not be simply regarded as a contraindication of radical resection.