Objective To investigate the role of IL-6 in the co-cultivation system of porcine hepatocytes with bone marrow mesenchymal stem cells (MSCs) in vitro.Methods Mononuclear cells were isolated from bone marrow aspirate of swines (n = 3) by 1.077 g/ml density gradient centrifugation.MSCs of passage 3 and primary hepatocytes harvested by a two-step in situ collagenase perfusion technique were inoculated in Millicell culture inserts at a seeding ratio of 1:2,and functional changes of heterotypic interactions were characterized.Levels of IL-6,TNF-α,and TGF-α were measured respectively.Results Hepatocyte viability was greater than 95%.Hepatocyte performance levels such as albumin secretion and urea synthesis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P＜0.05).The IL-6 level also significantly increased in co-culture group (P ＜ 0.01).A significant decrease of hepatic functions was observed upon neutralization of IL-6 in co-culture.Conclusion MSC-derived IL-6 plays a key role in the up-regulation of hepatocyte functions in this co-culture.

Objective To establish porcine hepatocyte co-culture system with bone marrow mes-enchymal stem cells (MSCs) in vitro for the ideal cell source of bioartificial liver. Methods Mononu-clear cells were isolated from bone marrow aspirate extracted from the anterior superior lilac spine of swines (n=3) by 1.077 g/ml density gradient centrifugation. MSCs of passage 3 and primary hepato-cytes harvested by a two-step in situ collagenase perfusion technique were randomly distributed, and the morphological and functional changes of heterotypic interactions were characterized. Results The purity of the third passage MSCs and primary hepatoeytes was more than 90 % and 99%, respectively.Hepatocyte viability was greater than 95 %. A rapid attachment and self-organization of three-dimen-sional hepatocyte spheroids were encouraged in co-culture. Heterotypic junctions remained similar to that of hepatocytes in vivo. Hepatocyte performance levels such as albumin secretion and urea synthe-sis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P<0.05). The best hepatic function levels were achieved on day 2 and moderately decreased in the following co-culture days. Conclusion Co-cultivation of porcine hepatocytes and MSCs may preserve hepatocyte morphology and function, which could contribute to the functional bioartificial liver.

Objective As an effective means of liver function support for acute liver failure, bioartificial liver has seen great progress in recent years. However, the development of this type of device is currently hindered by limited oxygen transport to cultured hepatocytes. In this study we try to resolve this problem by supplementing the circulating medium of the bioreactor with red blood cells.Methods Freshly isolated primary porcine hepatocytes were inoculated into our newly designed bioreactor and were divided into two groups: RPMI1640 was circulated in the control group and porcine red blood cells were added into the culture medium in the experimental group. The culture media in both groups were oxygenated through extra-corporeal membrane oxygenation, and the oxygen consumption rate (OCR) in the bioreactor was measured by a blood gas analyzer. Liver-specific functions and glucose consumption were also determined. Results The OCR of the experimental group was 1.5 fold that of the control group, and the glucose consumption rate was twice that of the control group. The liver-specific functions of the experimental group were significantly higher than those of the control group in terrns of albumin secretion and urea synthesis. Conclusion Supplementing the circulating medium of the bioreactor with red blood cells can significantly improve the oxygen supply in the bioreactor, thereby enhancing the glucose consumption and liver-specific functions of hepatocytes. This method is convenient and effective, and is expected to be an effective means to resolve the problems of oxygen supply in the bioreactor.

Objective To review the clinical experiences of modified Nuss procedure and technique for complicated pectus excavatum. Methods From September 2006 to October 2009,45 children (27 boys and18 girls), aged 3 - 16 years [mean (6.4 ± 3.5 )years] with complicated pectus excavatum underwent modified Nuss procedure. VATS was used in 6 cases. 3 patients associated with lung cyst were treated by open surgery and 1 case with ASD was treated by Hybrid procedure.Results The surgery was successfully performed in all patients. The mean operative time was 54 minutes and the average hospital stay was 7 days. Postoperative evaluation was"excellent"in 30 cases , "good"in 13, and"moderate"in 2. Conclusion Modified Nuss procedure is a safe and effective method for correction of complicated pectus excavatum.

Objective To investigate the clinical value of liver transplantation for intrahepatic cholangiocarcinoma (ICC).Methods A retrospective analysis was performed on 68 patients with ICC who underwent hepatic resection or liver transplantation between January 2007 and September 2013.In the surgical treatment of ICC,overall survival and prognostic risk factors were analyzed to determine the clinical value of liver transplantation for ICC compared with hepatic resection.Results Of the patients with ICC,55 underwent hepatic resection and 13 were given liver transplantation.The 1 and 3-year survival rate was significantly higher in the liver transplantation group than in the hepatic resection group (76.9％ versus 52.7％,and 61.5％ versus 27.3％,P＜0.05).In addition,liver transplantation resulted in higher tumor recurrence-free survival rate than hepatic resection (69.2％ versus 41.8％,and 61.5％ versus 20.0％,P＜0.05).The multivariate analysis showed that tumor size larger than 5 cm was an independent risk factor and HBV infection was an independent protective factor of ICC.Conclusion The prognosis of patients who underwent liver transplantation was better than hepatic resection,suggesting that liver transplantation is perhaps a prior treatment in strictly selected and unresectable patients with ICC.

ObjectiveTo study the effects of membrane molecular weight cut off on a novel bioartificial liver(BAL) system.MethodsHealthy beagles underwent 6-hour treatment with a BAL containing membrane with 200 kDa retention rating or 1200 kDa retention rating.The functional changes and cell viability were characterized.ResultsHepatocyte performance levels such as albumin secretion,urea synthesis and viability were significantly higher in the 200 kDa retention rating group when compared with the 1200 kDa retention rating group (P＜0.05).Significant levels of canine proteins were detected in the BAL medium from the 1200 kDa retention rating group.Fluorescence microscopy further verified that heavy deposition of canine IgG,IgM and complement (C3) on co-culture cells were obtained after BAL treatment in the 1200 kDa retention rating group.ConclusionsSmall membrane molecular weight cut off of BAL could reduce the transfer of xenoreactive antibodies into the BAL medium and improved the performance of the BAL.

Objective To investigate the influence of membrane molecular weight cut off in our bioartificial liver(BAL)system.Methods Beagle dogs were used for a model of acute liver failure through D-galactosamine administration.The acute liver failure Beagles were divided into two groups by the membrane molecular weight cut off.Group A was treated with BAL containing 200 kDa retention rating membrane.Group B was treated with BAL containing 1200 kDa retention rating membrane.Each group underwent two six-hour BAL treatments that were performed on day 1 and day 21.BAL medium were examined and levels of IgG,IgM,and complement hemolytic unit of 50％(CH50)antibodies were measured in all Beagles and.Results BAL treatment was associated with a significant decline in levels of CH50.1200 kDa group experienced a significant increase in levels of IgG and IgM after two BAL treatments.Significant levels of canine proteins were detected in BAL medium from 1200 kDa group.Conclusions Xenogeneic immune response in the BAL system was influenced by membrane molecular weight cut off.

Gallbladder cancer is a highly malignant tumor of the digestive system,and its incidence in China has been increasing year by year.Due to the lack of specific symptoms in the early stage,early diagnosis is difficult for gallbladder cancer and it is often at advanced stage once discovered.Surgery is still the only method to cure gallbladder cancer.Therefore,early diagnosis is particularly important for radical resection and improving overall prognosis of gallbladder cancer patients.However,there is still no specific molecular diagnostic target for gallbladder cancer.The continuous improvement of research and surgical skills have made the operational procedure of gallbladder cancer more standardized.With the increasing understanding on the epidemiology and pathogenesis of gallbladder cancer and the improvement of imaging,radiotherapy and chemotherapy technology,the comprehensive diagnosis and treatment of gallbladder cancer has also developed rapidly in recent years.However,high-quality clinical research is still needed.Comprehensive treatment model including radical surgery,adjuvant radiotherapy and chemotherapy,targeted therapy and immunotherapy will be the trend of gallbladder cancer treatment in the future.This article summarizes the advances in comprehensive diagnosis and treatment of gallbladder cancer by reviewing literatures.