Establishment of Humanized Mouse Model by Using Transplantation of Mobilized Peripheral Blood Stem Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the hematopoietic reconstitution in immunodeficiency NPG(TM) mice after transplantation of G-CSF-mobilized peripheral blood CD34(+) hemopoietic stem cells.</p><p><b>METHODS</b>CD34(+) cells were isolated from peripheral blood stem cells (PBSC) by magnetic activated cell sorting (MACS), and then were transplanted into NPG(TM) mice irradiated with sublethal dose of X ray by marrow cavity transplantation. The hemogram of mice after transplantation for 2, 4 weeks was observed; human cell populations (CD45(+), CD19(+)) in the peripheral blood of mice were dynamically analyzed by flow cytometry (FCM) at 4, 6, 8, 10 and 12 weeks after transplantation. Until the planned harvest at the 12 week after transplantation, the CD45(+), CD19(+) level in bone marrow, liver, spleen from each mouse were detected by flow cytometry; the expression of human Alu gene in the bone marrow cell of mouse was detected by PCR.</p><p><b>RESULTS</b>The purity of CD34(+) cells accounted for 96.3%; after irradiation, the nucleated cells and megalokaryocytes in the marrow cavity of NPG mice were reduced significantly or were lost, and reached the myeloablative effect. At week 4 after transplantation, components of blood cells in peripheral blood of transplanted mice were recovered to the level before irradiation; all the mice survived, human CD45(+), CD19(+) cells were found by FCM in the peripheral blood of all the surviving mice in transplantation group at week 4, 6, 8, 10, 12 after the transplantation; at the 12th week, the human Alu gene could be detected in the bone marrow of all the mice in transplantation group.</p><p><b>CONCLUSION</b>The human-mouse chimeric model is successfully established in irradiation-induced NPG mouse by transplantation of CD34(+) HSC from G-CSF-mobilized peripheral blood via marrow cavity.</p>

Morphology and microRNA expression profiles of drug-resistant cells in hepatocellular carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To compare morphological differences of three drug-resistant hepatocellular carcinoma (HCC) cell subclones (Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) and their parental Huh-7 cell line, to analyze differential microRNA (miRNA) expression profiles in these cells and, finally to screen for the abnormal expressed miRNAs in drug-resistant HCC cells.</p><p><b>METHODS</b>Cellular morphology was observed by histology and transmission electron microscopy. MiRNA microarray was used to analyze the differential miRNA expression profiles in these cells (Huh-7, Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) followed by real time quantitative PCR validation.</p><p><b>RESULTS</b>The drug-resistant cells had more intracytoplasmic organelles and were larger in size along with increased cytological pleomorphism than the parental Huh-7 cells. Compared with the parental Huh-7 cells, 32 simultaneously up-regulated and 22 down-regulated miRNAs were found in three drug-resistant cells. Up-regulation of miR-15a, miR-16, miR-27b, miR-30b, miR-146a, miR-146b-5p, miR-181a, miR-181d and miR-194 was verified by RT-qPCR.</p><p><b>CONCLUSION</b>Drug-resistant HCC cells have abnormal expressed miRNAs, which may be explored to further investigate the association of miRNA expressions with multidrugs resistance in HCC.</p>

Expression of survivin, p27 and PTEN in hepatocellular carcinoma and their clinical significances / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To detect the expression of survivin protein, survivin mRNA, p27 protein, p27 mRNA and PTEN protein in hepatocellular carcinomas (HCC) and their clinical significances.</p><p><b>METHODS</b>Tissue microarrays were constructed. The expression of survivin protein, p27 protein and PTEN protein were evaluated by immunohistochemical methods and in expression of survivin mRNA and p27 mRNA were evaluated by in stiu hybridization respectively in tumor tissues from 141 HCC patients, 128 samples of para-carcinoma liver tissues, 97 liver tissues far from the carcinomas and normal liver tissues from non HCC patients. The relationship of survivin, p27 and PTEN were investigated and a prediction model of HCC was constructed.</p><p><b>RESULTS</b>The expressions of survivin protein (Ridit 95% CI = 0.689+/-0.048, P < 0.01), survivin mRNA (Ridit 95% CI = 0.690+/-0.049, P < 0.01) and p27 protein (Ridit 95% CI = 0.556+/-0.053, P < 0.05) in HCC tissues were significantly increased, while the expression of PTEN protein (Ridit 95% CI = 0.282+/-0.048) in HCC tissues was significantly reduced (P < 0.01).</p><p><b>CONCLUSION</b>Overexpressions of survivin mRNA and p27 protein and reduced expression of PTEN protein might be a valuable marker to predict the presence of HCC.</p>