Passage of bone-marrow-derived liver stem cells in a proliferating culture system.

AIM: To explore the feasibility of passage of bone-marrow-derived liver stem cells (BDLSCs) in culture systems that contain cholestatic serum. METHODS: Whole bone marrow cells of rats were purified with conditioning selection media that contained 50 mL/L cholestatic serum. The selected BDLSCs were grown in a proliferating culture system and a differentiating culture system. The culture systems contained factors that stimulated the proliferation and differentiation of BDLSCs. Each passage of the proliferated stem cells was subjected to flow cytometry to detect stem cell markers. The morphology and phenotypic markers of BDLSCs were characterized using immunohistochemistry, reverse transcription polymerase chain reaction (RT-PCR) and electron microscopy. The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay. RESULTS: The conditioning selection medium isolated BDLSCs directly from cultured bone marrow cells. The selected BDLSCs could be proliferated for six passages and maintained stable markers in our proliferating system. When the culture system was changed to a differentiating system, hepatocyte-like colony-forming units (H-CFUs) were formed. H-CFUs expressed markers of embryonic hepatocytes (alpha-fetoprotein, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and cytochrome P450-2b1), and hepatocyte nuclear factors 1alpha and -3beta). They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: BDLSCs can be selected directly from bone marrow cells, and pure BDLSCs can be proliferated for six passages. The differentiated cells have hepatocyte-like phenotypes and functions. BDLSCs represent a new method to provide a readily available alternate source of cells for clinical hepatocyte therapy.

Modified technique of pancreaticojejunal anastomosis with invagination following pancreaticoduodenectomy: a cohort study.

BACKGROUND: Leakage from pancreatic anastomoses remains the single most important morbidity after pancreaticoduodenectomy and contributes to prolonged hospitalization and mortality. This observational cohort study reported the surgical outcome of a modified invagination technique of pancreaticojejunostomy after pancreaticoduodenectomy. METHODS: Between December 2001 and December 2007, a total of 52 consecutive patients underwent elective pancreaticoduodenectomy for benign or malignant pathologies of the pancreas or the periampullary region in a tertiary referral center. All patients underwent our modified invagination technique of pancreaticojejunostomy regardless of the characteristics of the pancreatic stump. Data were collected prospectively. RESULTS: The mean hospital stay was 12.6 +/- 3.2 days. The incidence of overall surgical complications was 9.6%. No patient developed pancreatic fistula. One patient (1.9%) died of respiratory failure on postoperative day 7. CONCLUSIONS: We reported our pancreaticojejunostomy anastomosis technique with a pancreatic fistula rate of 0% and low intra-abdominal complication rate. The favorable results of this technique warrant further investigation in large prospective cohort studies and prospective randomized controlled studies.

[Hand-assisted laparoscopic partial hepatectomy in the treatment of primary liver cancer].

OBJECTIVE: To explore the techniques of hand-assisted laparoscopic partial hepatectomy and its clinical value in the treatment of primary liver cancer. METHODS: Six patients with primary liver cancers in the segments II, III, V, VI and on the edge of the liver received laparoscopic partial hepatectomy assisted by special instruments such as hand-assisted devices, harmonic scalpel, and Endo-GIA. RESULTS: All of the 6 operations were completed successfully, in which resections of both II and III segments, VI segment, and non-regular segment were performed. The operative time was 54 to 130 min and postoperative hospital stay 6-9 days, with intraoperative blood loss of 150-700 ml. No conversion to laparotomy was needed, nor did any complications occur. Postoperative follow-up lasted for 6 to 15 months, during which relapse occurred in 2 cases 3 months and 4 months after the operations respectively, while no recurrence has been found in the other 4 patients. CONCLUSION: Hand-assisted laparoscopic partial hepatectomy is safe and feasible for primary liver cancer in clinically selected patients.

[Postoperative complications after orthotopic liver transplantation: clinical analysis of 18 cases].

OBJECTIVE: To investigate experience in diagnosis and treatment of postoperative complications in patients undergoing orthotopic liver transplantation (OLT). METHODS: Complications, treatment and management following liver transplantation in 16 cases were analyzed retrospectively. RESULTS: Of 16 patients, 5 patients had advanced liver cirrhosis, 7 primary liver carcinoma, 1 liver failure after hepatectomy for liver cancer, 1 Wilson's disease, 1 chronic renal failure and liver cirrhosis and 1 acute live failure. Twelve patients survived, the longest survival was 4 years. Complications following OLT included: intra-abdominal bleeding in 3 cases, intracerebral vascular lesions in 2, pulmonary infection in 6, adult respiratory distress syndrome in 2, suprahepatic inferior vena caval occlusion in 2, hepatic artery thrombus in 1 case, bile duct stone or sludge in 3, bile leakage in 1 case, acute rejection in 2, chronic rejection in 2, acute renal failure in 2. Six patients died during perioperative period, one patient died of intracerebral bleeding, one from adult respiratory distress syndrome, one of acute renal failure one of hepatic artery thrombus, one of acute rejection and one of liver failure. CONCLUSIONS: Proper prevention and treatment can effectively reduce complications following OLT during perioperative period. The timely diagnosis, treatment and prophylactics are necessary to prevent these complications.

[Clinical experience with 13 cases of orthotopic liver transplantation].

OBJECTIVE: To review our experience with orthotopic liver transplantation. METHODS: Thirteen liver transplantation were performed in 12 patients (including one liver retransplantation), of whom 5 patients received the transplantation for end stage liver cirrhosis, 4 for primary liver carcinoma, 1 for liver failure after hepatectomy for liver cancer, 1 for Wilson's disease, and 1 for chronic renal failure and liver cirrhosis. Retransplantation was done in 1 patient for chronic graft rejection. Of the 13 operations, 10 underwent the classical procedures of orthotopic liver transplantation, while 2 adopted modified piggyback technique, with 1 of the patients receiving retransplantation. RESULTS: Nine patients survived the transplantation with the longest survival over 2 years. Four patients died in the perioperative period, due to intracerebral bleeding, adult respiratory distress syndrome, acute renal failure and hepatic artery thrombus, respectively. CONCLUSIONS: Liver transplantation is an effective treatment for various end-stage liver diseases. Strict patient selection, appropriate timing of the operation and proper perioperative care are all essential for the success of liver transplantation.