BACKGROUND:Engineered hepatic tissue is considered a promising strategy for healing acute liver failure. But, there are series of hindrances in the construction of engineered hepatic tissues, including acquisition of vital hepatocytes, choice of scaffolds and culture system, and nutrition supply. OBJECTIVE:To construct three kinds of engineered hepatic tissues in hope to screen the optimal one for transplantation in acute liver failure. METHODS:After purification, amplification, bone marrow mesenchymal stem cells were induced to differentiate into hepatocyte-like cells which were co-cultured with acellular amniotic membrane, DACRON PATCH cardiovascular surgical patch, biological surgical patch, respectively to construct three kinds of engineered hepatic tissues. After 3 days of culture, morphological and functional detections were performed. RESULTS AND CONCLUSION:Rat bone marrow mesenchymal stem cells with higher purity were successful y harvested by using density gradient centrifugation and adherent methods, and then the cells were differentiated into hepatocyte-like cells. In the three kinds of engineered hepatic tissues, hepatocyte-like cells were found to be combined with the biological surgical patch to the maximum extent, and their combination exhibited stronger ability of urea synthesis and albumin secretion, which provides experimental basis for treatment of acute liver failure.

Objective To Study the effect of splenectomy plus endoscopic varices ligation (SEVL) in the treatment of portal hypertension. Methods SEVL was carried out in 38 patients with portal hypertension from January 2000 to June 2002. The authors compared hemodynamic parameters and gastric emptying time before and after the operation. The incidence of upper gastrointestinal bleeding, the disappearance rate of esophageal varicosis, the outcomes of portal hypertensive gastropathy (PHG) and the incidence of complications were recorded at postoperative 3 weeks, 6 months, 12 and 18 months, respectively. Results Postoperatively, the free portal pressure, portal flow volume and velocity were significantly reduced in all the cases ( P 0.05). Portal thrombosis was seen in 6 cases (6/38, 15.8%). Conclusions SEVL has little influence on gastric emptying and PHG, offering a high disappearance rate of varicosis and a low incidence of complications.

Objective To investigate the expression and significance of ?-endorphin in hippocampus after high power microwave (HPM) radiation. Methods 50 male Wistar rats were sacrificed at 6h, 1d, 3d, 7d, 14d, 28 days after exposure to 12mW/cm~2 HPM imitational source radiation. The hippocampus was harvested, the characteristics of injury to the hippocampus and expression of ?-endorphin were evaluated by means of light microscope, EM, immunohistochemistry and image analysis. Results In the neutrons of hippocampus, mitochondrial swelling and myelin sheath confluence and dissociation were observed 6 hours after radiation, and mitochondrial swelling, cavitation, disruption of crista, concentration margining of chromatin, blurring of synaptic cleft, piling or evacuation of vesicles were observed 3 days after HPM radiation. The expression of ?-endorphin in cytoplasm of neurons was up-regulated continuously from 1 to 7 days, peaking at 7 days, restoring to normal level from 14 to 28 days after radiation. Conclusion HPM can produce injury to the hippocampus at histological and ultrastructural levels. The expression of ?-endorphin is up-regulated, and it might play an important role in the pathophysiological process of injury and repair in the hippocampus.

Objective To evaluate the effect of endoscopic variceal ligation(EVL) combined with splenectomy for the patients with portal hypertension. Methods Thirty two cases of portal hypertension underwent EVL plus splenectomy from May 1999 to May 2002 in our department. The patients were registered and followed up in the third, sixth and twelfth months after operation respectively. Re ligations were performed in case of relapsed varices. Results Thirty one patients were followed up for ≥2 years.Of them,esophageal varices recrrented in 6.4% of patients during following-up period,but no reblecding, and no death caused by hepatic encehpalopathy or other complications.Conclusions EVL combined with splenectomy has less trauma , less postoperative complications and high eradication rate of esophageal varices, so it can be used safely in the treatment of patients with ruptured esophageal varices.

AIM:To investigate morphologic and functional changes of small intestinal mucosa and proliferating cell nuclear antigen in postoperative portal hypertension patients with single or combined administration of Gln and rhGH.METHODS:Twenty-nine portal hypertension patients with surgical treatment were prospectively randomized to four groups as follows:① Gln group(n=6);② rhGH group(n=8);③ Gln+rhGH group(n=7)and ④ control group(n=8).A standard solution for TPN was given three days after operation for a week.The concentration ratio of urinary lactulose and mannitol(L/M),the villus height and crypt depth and PCNA index of small intestinal mucosa were compared.RESULTS:A week after TPN postoperation,the increased ratios of L/M in Gln+rhGH group were less than those in control group(P0.05).CONCLUSION:This study suggest that Gln together with rhGH reduce the intestinal permeability and protect the mucosa integrality in postoperative portal hypertension patients,but not in single treatment.

Objective To investigate the distribution and changes of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in normal heart or heart after myocardial infarction (MI),and to explore beneficial effects of bFGF on heart after MI. Methods Eleven adult minipigs were randomly divided into 2 groups,control (n=5) and MI group (n=6). MI was induced by ligating the left anterior descending coronary artery. Sham-operation was carried out on the animals of control. bFGF-like-immunoreactivity (bFGF-ir) and FGFR-1-like-immunoreactivity (FGFR-1-ir) in heart tissues were detected by immuno-histochemistry and image analysts in 4 weeks after surgery. Results In controls,bFGF-ir and FGFR-1-ir in the atrium showed a considerable high level compared with 2 ventricles (P

BACKGROUND: There is no accepted treatment for liver fibrosis recently. Bone marrow meaenchymal stern cells (BMSCs) used in the treatment of liver fibrosis has been reported as an effectively treatment, but the mechanism is unclear.OBJECTIVE: To study the regulation of hepatic stellate cells mediated by human BMSCs in vitro.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Center for Stem Cells and Tissue Engineering of Sun Yat-sen University and the Central Laboratory of Third Affiliated Hospital of Sun Yat-sen University from June to December 2008.MATERIALS: Human bone marrow masenchymal stem cells were collected from normal youth volunteers; Human hepatic stellate cells and normal liver call line L-O2 were supplied by the Animal Experimental Center of Sun Yat-sen University.METHODS: The purified human BMSCs and hepatic stellate calls were set up in Transwell co-culture system. The incubation density was 2×104cells/well. L-O2 was set up instead of human BMSCs as negative control. Hepatic stellate cells cultured alone served as blank control group. The culture was performed for 72 hours.MAIN OUTCOME MEASURES: Morphology of hepatic stellate cells and results of immunocytochemical staining. Apoptosis of hepatic stellte calls was determined by flow cytometry. Western blot were used to assay the expression of α-actin.RESULTS: Activated hepatic stellate cells presented fiat and thin shape under an inverted microscope. Fat drop was lack in cytoplasm, a -actin located in hepatic stellate calls, with the presence of high tension fibers. Compared with the L-O2 + hepatic stellate cell and hepatic stellate call groups, the apoptotic rate of hepatic stellate cells was significantly increased in the BMSC + hepatic stellate cell group (P < 0.05). α -actin expression was significantly down-regulated.CONCLUSION: Human BMSCs can inhibit activation of hepatic stellate ceils and promote them apoptosis, which may be the anti-hepatic fibrosis mechanism of BMSCs.

Objective To evaluate HCC family history on recurrence and survival of HCC patients after curative hepatectomy. Methods The family history,postoperative recurrence rate,1,2 and 5 year survival rate of 79 HCC patients who received surgical resection were collected.Patients were divided into two groups:34 cases in familial aggregation HCC group (FH group ) and 45 cases in non- familial aggregation HCC group (NH group).All these HCC patients were followed up for 3 months to10 years after surgery,and the median follow-up period was 48.5 months.We analyzed the survival rate of HCC patients by Kaplan -Meier survival curve to compare the postoperative recurrence rate and survival rate between two groups. Results 34 cases of HCC probands came from 33 families,and there were 104 cases of HCC patients in these families,an average of 3.2 cases per family.There were 6 families in which 3 or more cases developed HCC,and there were 13 families in which father or (and) mother had HCC accounting for 39.4％.The six months,1 year,2 year posthepatectomy recurrence rate in FH group was 15.1％,22.3％,40.1％,significantly higher than NH group of 10.3％,17.4％,25.6％ (x2 =5.762,6.434,5.987,P ＜0.05),and 1,2,5 year's survival rate in FH group was 57％,46％,40.3％,much lower than NH group of 85.3％,75.2％ and61.5％ (x2 =8.986,9.765,7.634,P＜0.05). Conclusions Family history of HCC affects the prognosis of HCC patients after surgery. Patients with HCC family history have higher postoperative recurrence rate and worse prognosis than that of patients who do not have HCC family history.

BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.

Objective To construct the bioartificial liver by bone mesenchymal stem cell (BMSC) and alginate scaffold. Method Alginate scaffold was used as the cell carrier for the cultivation of BMSC and the differentiation from BMSC into hepatic like cells was induced by the cell factors of HGF, EGF and FGF-4 in the scaffold in vitro. The compatibility of the cells and the scaffold was observed by microscopy and the function of the differentiatd cells was tested. The gene of AFP and ALB was detected by RT-PCR. The secretion of ALB and the urea synthesis of the cells were tested by ALB kit and urea kit respectively. The glycogen synthesis and the CK-18 was tested by the glycogen stanning method and the immunofluorescence test. Results BMSC was able to attach, grow and proliferate well in the alginate scaffold, the well compatibility was observed by microscopy. ALB and urea were detected in the cultivating medium, the gene of ALB and AFP was identified by RT-PCR. The glycogen synthesis ability and the expression of CK-18 were induced during the differentiation. Conclusion The three dimensional atginate scaffold exhibited well compatibility with BMSC, BMSC could be differentiated into the hepatic like cell in the scaffold. BMSC and the alginate scaffold could be used to construct the bioartificial liver for the hepatic tissue engineering.