Objective To establ ish a new induction method from human embryonic stem cells (hESCs) differentiating into hepatocyte-l ike cells using an adherent culture system with single-step induction. Methods Undifferentiated hESCs were cultured on Matrigel-coated culture plates for 4 days, hepatic differentiation was initiated at 60%-70% confluence by adding Activin A for 5 days. Then the induction medium was replaced by hepatocyte induction medium (HIM) supplemented with fibroblast growth factor 1 (FGF-1) and bone morphogenetic protein 4 (BMP-4) for another 6 days. Finally, the cells were treated with HIM adding hepatocyte growth factor (HGF) and Oncostatin M (OSM) for 5-7 days. The characteristics of differentiated cells were determined by morphology, immunofluorescence staining, RT-PCR, and Periodicacid- Schiff (PAS) test. Results Differentiated cells treated with Activin A, FGF-1, BMP-4, HGF, and OSM sequentially were morphologically larger and became spherical, oval or polygon. Some cells had 2 or 3 nuclei, suggesting that the cells have a hepatocyte-l ike morphology. Differentiated cells at first induction stage could be stained positive by SOX17 and Forkhead (FOX)A2 after induction by Activin A. Then they turned to be α fetoprotein (AFP) and α1 antitripsin positive cells at second induction stage after induction by FGF-1 and BMP-4. Finally, the differentiated cells treated with HGF and OSM showed PAS possitive for glycogen detection. The differentiated cells at various stages also expressed at early (SOX17, FOXA2, and GATA-4),middle (AFP, albumin, and cytokeratin 18), and mature (alcohol dehydrogenase 1C and Cytochrome P4501B1) stage hepatic genes, respectively. Conclusion Using a simple-step induction method and by suppl ied with cytokines consequently, hESCs can be induced to differentiate into hepatocyte-l ike cells. The differentiation method can provide seed cells for hepatic tissue engineering or cell-therapy.
ObjectiveTo study the effects of recombinant human growth hormone (rhGH) on proliferation of human rectal cancer cell in vitro. MethodsThe experiment was divided into control group,rhGH group,Oxaliplatin (LOHP) group and rhGH+LOHP group. The double proliferation time of cells,cell inhibition rate,cell cycle, proliferation index (PI) and DNA inhibition rate of human rectal cancer line,HR8348,were studied by cell culture, MTT assay and flow cytometry on different concentration of rhGH. ResultsIn vitro the markedly accelerated effects of rhGH on multiplication of HR8348 cell line were not found: there was no statistical significance as compared rhGH group with control group or compared rhGH+LOHP group and LOHP group （Pgt;0.05）. The double proliferation time of cells was markedly lengthened, cell inhibition rate and the cells arrested in G0-G1 phase were obviously increased, meanwhile, the cells in S phase （P＜0.05） and G2-M phase and PI were markedly decreased and DNA inhibition rate was obviously risen as compared rhGH+LOHP group with control group or rhGH+LOHP group and rhGH group （P＜0.01）.ConclusionIn vitro rhGH does not accelerate the multiplication of human rectal cancer cells.
Objective To study the effects of edaravone on the lung injury of severe acute pancreatitis （SAP） in rats. Methods Thirty-six SD rats were randomly divided into three groups: normal control group, model group and edaravone group, and SAP was induced by intraductal administration of 5% sodium taurocholate. Edaravone was given in edaravone group, while normal saline was given in normal control group and model group. After operation 6 h rats were executed, and dry/wet weight (D/W) ratio of lung was counted, and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity in serum and lung were detected, respectively. In addition, the levels of tumor necrosis factor-α (TNF-α), interleukin-1, -6 (IL-1, -6) of serum were detected.Results The MDA contentof serum and lung and the levels of TNF-α, IL-1, IL-6 in model group were markedly higher than those in normal control group and edaravone group, but D/W ratio of lung, SOD activity of serum and lung were significantly lower (Plt;0.05). Conclusion Edaravone can alleviate lung injury of rats caused by SAP.
Objective To investigate the expression of growth hormone receptor (GHR) in human gastric cancer tissue. Methods The GHR was detected in samples of the human gastric cancer (57 cases) and the distal normal tissues (57 cases) by immunohistochemistry technique. Results The GHR expression positive rate was 80.7%(46/57) in the human gastric cancer tissues and 70.2%(40/57) in the distal normal tissues. There was no statistic difference between the human gastric cancer tissues and the distal normal tissues (Pgt;0.05). There were also no statistic differences among the gastric cancer tissues of different differentiation, different tissue type, different gender and different age ranges (Pgt;0.05). Conclusion It is similar that the expression of GHR between the human gastric cancer tissues and the distal normal tissues.
Objective To investigate the effect of recombinant human growth hormone (hGH) on growth situation and bone marrow hematopoietic function in rats under chemotherapy. Methods A total of 136 10-week-old SD rats were included in the study. The rats were randomly assigned into five groups: normal control group (n=8), normal saline control group (NS group, n=32), human growth hormone group (hGH group, n=32), chemotherapeutic drug treated group (CT group, n=32), and chemotherapeutic drug plus hGH treated group (CT+hGH group, n=32). The body weight, bone marrow differential count, and the expression of proliferating cell nuclear antigen (PCNA) in bone marrow were measured before treatment and weekly in four weeks after treatment. Results Weight loss occurred in both CT group and CT+hGH group (P＜0.05), but the weight loss in CT+hGH group was significantly smaller (P＜0.05) on day 7 after treatment. In myeloid morphology, myeloid cell was hypoplastic excessively in CT group, and it was hypoplastic obviously in CT+hGH group on day 7 after treatment. Day 14, weight gain appeared in CT+hGH group, while weight loss remained in CT group; In myeloid morphology, myeloid cell was hyperplastic actively excessively in CT+hGH group, and myeloid karyote count was increased significantly in CT+hGH group (P＜0.05). Day 7, 14 and 21, PCNA positive cells count in CT group was lower than that in hGH group and CT+hGH group (P＜0.05). There was no significant different of every index among normal control group, NS group, and hGH group (Pgt;0.05), except weight between normal control group and hGH group on day 7 (P＜0.05). Conclusion hGH has a protective effect on myeloid hematopoietic function and growth situation in rats after intraperitoneal chemotherapy.