Adult-to-adult living-related donor liver transplantation: report of 2 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the feasibility and safety of adult-to-adult living-related donor liver transplantation using a right lobe graft.</p><p><b>METHODS</b>The clinical data of 2 cases of living-related donor liver transplantation performed between July, 2010 and November, 2010 were analyzed.</p><p><b>RESULTS</b>Liver transplantation was performed using a right lobe graft including the middle hepatic vein in one case and a right lobe graft without the middle hepatic vein in the other. The ratio of graft volume to standard liver volume was 46.2% and 47.3% in the two cases, with GR/WR of 0.83 and 0.80, and donor residue liver of 42.1% and 39.5%, respectively. The donor operation lasted for 6.5 h and 5 h in the two cases with blood loss of about 200-250 ml without blood transfusion. The donors recovered uneventfully without any surgical complications, whose liver function was normal 7 days after the operation, and were discharged 14 days and 16 days after the surgery, respectively. The recipient operation lasted for 8 h and 7 h with blood loss of about 800-1000 ml. The right hepatic vein, hepatic artery, portal vein and bile duct reconstruction were performed by end-to-end anastomoses in the 2 recipients. Bile duct anastomosis stricture occurred in the first recipient 2 months after transplantation and was treated with percutaneous transhepatic cholangiography and drainage. The second recipient recovered smoothly without any complications. The recipients have so far survived 9 months and 5 months, respectively.</p><p><b>CONCLUSION</b>Adult-to-adult living-related donor liver transplantation is a safe and effective option for treatment of end-stage liver diseases in the context of cadaveric liver graft shortage.</p>

Variations of the amount of sialic acids on hepatocellular carcinoma cell membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the change in the amount of sialic acids on hepatocellular carcinoma (HCC) cell membrane.</p><p><b>METHODS</b>Surgical specimens of HCC and liver cirrhosis tissues were obtained from 28 patients to prepare carcinoma cell and hepatocyte suspensions by collagenase digestion. For assay of α2, 3 and α2, 6-sialic acids, the cells were suspended in the staining buffer containing either fluorescein isothiocyanate-Maackia amurensis lectin (FITC-MAL) or fluorescein isothiocyanate-Sambucus nigra bark lectin (FITC-SNA) and incubated for 1 h, respectively. Flow cytometric analysis was carried out to measure the mean fluorescence intensity (MFI) on the cell surface.</p><p><b>RESULTS</b>In both FITC-MAL- and FITC-SNA-incubated HCC cells, the MFI on the cell surface was greater than that of the hepatocytes.</p><p><b>CONCLUSION</b>Both of α2, 3 and α2, 6- sialic acids increases significantly on the hepatocyte membrane after the carcinomatous change.</p>

Intrahepatic transplantation of in vitro induced autologous bone marrow-derived liver stem cells in patients with posthepatitic cirrhosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of in vitro induced autologous bone marrow-derived liver stem cell transplantation for posthepatitic cirrhosis.</p><p><b>METHODS</b>Between Jun 2008 and Mar 2009, 12 patients with posthepatitic cirrhosis and portal hypertensive underwent azygousportal disconnection and splenectomy in our department. The patients were then divided into two groups to receive autologous bone marrow-deprived liver stem cell infusion via the hepatic artery after in vitro induction for 7 days (n=6) or saline (n=6). The therapeutic effects of the operations on the liver functions and liver fibrosis index were evaluated.</p><p><b>RESULTS</b>All the patients recovered uneventfully and no side effect of the operation was found. After the operation, the patients receiving bone marrow-deprived liver stem cell infusion showed better hepatic function improvement than those receiving saline infusion (P<0.05).</p><p><b>CONCLUSION</b>Transplantation of in vitro induced autologous bone marrow-derived liver stem cell via the hepatic artery is safe and effective for treatment of posthepatitic cirrhosis.</p>

Protocols for cloning human bone marrow-derived hepatic stem cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore practical protocols for cloning bone marrow-derived hepatic stem cells in vitro.</p><p><b>METHODS</b>The cell fraction rich in CD117(+) cells and CD184(+) cells was separated from fresh bone marrow by density gradient centrifugation and cultured for 0, 7 and 14 days in high-glucose DMEM supplemented with or without 10% autologous serum or in serum-free high-glucose DMEM. All the media were supplemented with different concentrations of hepatocyte growth promoting factors (HGPF), thrombopoietin (TPO) and interleukin-3 (IL-3). The quantitative changes of CD117(+) cells and CD184(+) cells were measured by flow cytometry.</p><p><b>RESULTS</b>The optimal effect for cell cloning was achieved with high-glucose DMEM with 10% autologous serum group supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3. At day 7 of cell culture in this media, the quantity of CD117(+) cells and CD184(+) cells increased by 6.55 and 6.20 folds, and by 11.62 and 20.57 folds at day 14, respectively.</p><p><b>CONCLUSION</b>It is practical for cloning bone marrow-derived hepatic stem cells in high-glucose DMEM with 10% autologous serum supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3.</p>

Establishment of a mouse model of biliary obstruction and its dynamic observations / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a mouse model of biliary obstruction.</p><p><b>METHODS</b>Sixty-four Balb/c mice were divided into experimental group and control group. Obstructive jaundice was induced in the mice in the experimental group by common bile duct ligation. The level of the common bile duct diameter, WBC, LYM MID, LYM%, MID% and ALT, AST, TBIL, DBIL, IBIL, ALP and CHOL were measured 12 h and 1, 2 ,3, 4, 5, and 7 days after the ligation. The morphological changes in the liver were also observed.</p><p><b>RESULTS</b>The level of common bile duct diameter, WBC, LYM, MID, LYM%, MID% and ALT, AST, TBIL, DBIL, ALP and CHOL all underwent changes with time following certain patterns.</p><p><b>CONCLUSION</b>The jaundice manifestation of this model is similar to that of patients with biliary obstruction, and this model may provide a reliable model for studying the mechanism of obstructive jaundice.</p>

Effects of hepatic growth factor on c-kit+Lin- cell proliferation in mouse bone marrow / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine the optimal cytokine combinations with hepatic growth factor (HGF) that results in the most significant simultaneous in vitro expansion of cc-kit(+)Lin(-) cells derived from the bone marrow.</p><p><b>METHODS</b>C-kit(+)Lin(-) cells were isolated from mouse bone marrow using a high-gradient magnetic cell sorting system (MACS) and expanded in the presence of stem cell factor (SCF), FLt-3 ligand (FL), leukemia inhibitor factor (LIF) thrombopoietin (TPO) and different concentrations of HGF for 7days in a liquid culture system. The total cell number and Annexin-V-positive cell number were counted, and the antigen expressions were studied with fluorescence-activated cell sorting (FACS).</p><p><b>RESULTS</b>In each group, c-kit(+)Lin(-) cells were expanded effectively and rapidly by 2 to 8 folds. Addition of 10 ng/ml HGF into SCF+FL+LIF+TPO resulted in the most significant expansion of c-kit(+)Lin(-) and total cells by 8.00 and 45.43 folds, respectively, with cell apoptosis rate of 17.42 %. But as the concentration of HGF increased, the c-kit(+)Lin(-) cells and the apoptosis rate decreased.</p><p><b>CONCLUSION</b>HGF at10 ng/ml shows optimal synergistic effect with SCF, FL, LIF and TPO in expansion of c-kit(+)Lin(-) cells, and excessive HGF may induce cell differentiation.</p>

Differentiation of transplanted mouse c-Kit+lin- bone marrow cells into hepatocytes in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To ascertain whether mouse c-Kit(+)Lin- bone marrow cells have the potential of hepatic stem cells.</p><p><b>METHODS</b>c-Kit(+)lin- bone marrow cells were isolated and purified by magnetic-activated cell sorting (MACS) from BALB/C male donor mice, and immediately transplanted into age-matched BALB/C syngeneic female mice with 35-Gy total liver irradiation. The recipients were sacrificed 1 month after the transplantation for pathological observation of the liver morphology. The presence of Y-chromosome was examined in the liver cells of the recipient by in situ hybridization (ISH), and alpha-fetoprotein (AFP) and albumin in the cells were detected by immunohistochemistry.</p><p><b>RESULTS</b>The hepatocytes positive for Sry gene on Y-chromosome were identified 1 month after transplantation, and immunohistochemistry for AFP and albumin confirmed that the donor mice-derived cells were hepatocytes.</p><p><b>CONCLUSION</b>c-Kit(+)lin- bone marrow cells have the potential of hepatic stem cells, which can reside and differentiate into hepatocytes in the liver after transplantation. c-Kit(+)lin- bone marrow cells can be used as the source cells of cell transplantation for liver disease.</p>