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BACKGROUND: Periosteal cells are precursors of osteoblasts and chondrocytes. Some studies have reported that bone morphogenetic protein-7 can be used to induce periosteal cell proliferation, but limited by the high cost. Phytoestrogen icariin-induced periosteal cell proliferation has provided a new direction for tissue engineering. OBJECTIVE: To investigate the effect of icariin on the proliferation of human periosteum cells and to analyze the underlying mechanism. METHODS:Human periosteal cells were cultured in vitro and seeded to the 24-well plate with the concentration of 103/well after third passage. Cell proliferation test: the cells were cultured in the cell culture medium (control group), or the culture medium containing different concentrations of icariin (10-1, 10-2and 10-3mg/L). Proliferation mechanism test: the cells were cultured in the cell culture medium (control group), or the culture medium containing different concentrations of icariin (10-1, 10-2and 10-3mg/L) plus estrogen receptor antagonist ICI182.780. The cell proliferation in each test was detected by MTT assay at 1, 2 and 3 days of culture. The effects of different concentrations of icariin on the levels of estrogen receptor α and β proteins in the periosteal cells were detected by western blot assay. RESULTS AND CONCLUSION:The proliferation of human periosteum cells in vitro was successful,and icariin with the concentrations of 10-1, 10-2and 10-3mg/L could significantly the cell proliferation (P < 0.05). However, this effect was blocked after ICI182780 addition (P < 0.05), and the levels of estrogen receptor α and β were upregulated. To conclude, icariin can enhance the proliferation of periosteal cells probably by upregualting the expression of estrogen receptor α and β.
RESUMO
<p><b>OBJECTIVE</b>To analyze the characteristics of CDR3 of TCRbeta on CD8+ T cells in chronic hepatitis B patients.</p><p><b>METHODS</b>Eight patients with chronic hepatitis B (ALT more than 2 ULN) were enrolled in this study. CD8+ T cells were isolated from peripheral blood. RT-PCR was proformed to amplify the CDR3 of TCRbeta, and the PCR products were sequenced and analyzed.</p><p><b>RESULTS</b>The chronic hepatitis B patients showed obvious clonal expansion of T cell, and three perturbation patterns of T cell expansion were showed in the CDR3 of TCRbeta, including monoclonicity, oligoclonicity and skewed peak patterns. The number of perturbation families of CD8+ subpopulation was significantly higher than that of CD8- subpopulation (10.6+/-4.7 vs. 4.1+/-3.1, t = 6.619, P less than 0.01). In 3 out of 8 patients, the number of perturbation families of CD8+ subpopulation was also higher than that of PBMCs without depleting CD8+ subpopulation.</p><p><b>CONCLUSIONS</b>The characteristics of CDR3 of TCRbeta may help to understand the inflammatory response in CHB patients.</p>