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>

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>

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>

Identification of necrophagous fly species from 12 different cities and regions in China using inter-simple sequence repeat melocular markers / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify necrophagous fly species from different regions in China using inter-simple sequence repeat (ISSR) melocular markers and analyze their genetic difference and relationship.</p><p><b>METHODS</b>Five carrion fly species were collected from 12 cities and regions in China, including M.domestica, Lucilia sericata, Chrysomyia megacephala, Helicophagella melanura, and Boetthcherisca peregrina. Twenty-two ISSR primers were designed and synthesized, from which 8 were selected to identify the necrophagous fly species. Cluster analysis was conducted based on distance matrices using unweighted pair group method.</p><p><b>RESULTS</b>Totally 121 amplification samples were obtained using the 8 primers, and 679 clear and stable bands were visualized including 516 bands with polymorphisms. M.domestica, Lucilia sericata, Chrysomyia megacephala, Helicophagella melanura, and Boethcherisca peregrina from different regions in China produced their specific PCR band spectra. M. domestica from 10 different regions in China showed different inheritance patterns of the markers. Species-specific ISSR fragment was found among the necrophagous flys pecies. Cluster analysis among the most abundant carrion fly species demonstrated that M.domestica from 10 different regions could be divided into 4 groups at different levels. Most of the Chrysomyia megacephala and Lucilia sericata could be clustered in one tree.</p><p><b>CONCLUSION</b>This study represents the first identification of the common necrophagous fly species in China. ISSR-PCR-based identification of the species reveals the genetic diversity and genotypic difference among M.domestica from 10 cities and regions in China.</p>