ISSR analysis of genetic diversity and relationship among medicinal plants of Epimedium acuminatum from different habitats in Guizhou Province / 中草药

Objective: To analyze the genetic diversity and relationship of Epimedium acuminatum from different habitats in Guizhou Province. Methods: PopGene 32 software and NTsys2.10e software was used to analyze the genetic diversity and phylogenetic relationship of five different habitats in Guizhou Province, and phylogenetic tree was constructed according to the UPGMA method. Results: A total of 13 polymorphic and clear bands were screened from 100 random primers showed that amplified 211 bands, of which 205 were polymorphic from the level of species, the percentage of polymorphic bands (PPB) was 97.16%. The Shannon’s information index (I) was 0.455 7; Nei’s gene diversity index (He) was 0.297 3; The effective number (Ne) of alleles was 1.490 2; The total genetic diversity (Ht) of five populations was 0.297 3, while the population genetic diversity within the group (Hs) was 0.207 5, indicated that the genetic variation mainly existed within populations, genetic differentiation within population than that of population between. The population genetic distance clustering was built by UPGMA. The results showed that five populations were divided into two branches, three populations from Anshun, Huaxi, and Gaopo clustered into one branch, and the other two populations from Longli and Leishan populations clustered into one branch. Conclusion: ISSR molecular marker technique can be used to analyze the genetic diversity and phylogenetic relationship of Epimedium acuminatum from different habitats in Guizhou Province.

Inhibition of Combination of Icaritin and Doxorubicin on Human Osteosarcoma MG-63 Cells in vitro / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the inhibition and molecular mechanism of icaritin (ICT) combined doxorubicin (DOX) on human osteosarcoma MG-63 cells in vitro.</p><p><b>METHODS</b>The control group, ICT groups (10, 20, 40, 80, and 160 µmol/L), DOX groups (1, 2, 4, 8, and 16 µg/mL), and combination groups (20 µmol/ L ICT +1 µg/mL DOX, 20 µmol/L ICT +2 µg/mL DOX, 20 µmol/L ICT +4 µg/mL DOX, 40 µmol/L ICT +1 µg/mL DOX, 40 µmol/L ICT +2 µg/mL DOX, 40 µmol/L ICT +4 µg/mL DOX, 80 µmol/L ICT +1 µg/mL DOX, 80 µmol/L ICT +2 µg/mL DOX, 80 µmol/L ICT +4 µg/mL DOX) were set up. Human osteosarcoma MG-63 cells were respectively cultured and their effects on morphological changes were observed using inverted phase contrast microscope after 24-and 48-h intervention. The cell proliferation inhibition rate of each group was de- termined using CCK-8, and IC50 calculated. The MG-63 apoptosis rate was detected using Annexin V-FITC/ PI double dye flow cytometry. Expression levels of bcl-2, caspase-3, and p21 were detected using RT-PCR.</p><p><b>RESULTS</b>ICT and DOX could obviously inhibit the proliferation of MG-63 cell. Along with ICT concentration increasing from 10 µmol/L to 160 µmol/L, the cell proliferation inhibition rate also increased gradually from 9.67% ± 3.62% to 89.18% ± 9.66%. The IC50 was 46.93 µmol/L and 3.87 µg/mL respectively. ICT and DOX could cause either early or late stage apoptosis, down-regulate Bcl-2 gene expression, and up-regulate gene expressions of Caspase-3 and p21 respectively (P < 0.05). Aforesaid changes were more obviously seen in combination groups than in lCT groups and DOX groups (P < 0.05).</p><p><b>CONCLUSION</b>CT combined DOX had additive or synergistic inhibition effect for the proliferation of osteosarcoma MG-63 cells, which might be related with regulating gene expressions of bcl-2, caspase-3, and p21.</p>

Effects of Leukemia Inhibitory Factor Combined with Basic Fibroblast Growth Factor on Self-maintenance and Self-renewal of Human Umbilical Cord Mesenchymal Stem Cells In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effects of LIF combined with bFGF on the proliferation, stemness and senescence of hUC-MSC.</p><p><b>METHODS</b>Experiments were divided into 4 groups: control group, in which the cells were treated with complete medium (α-MEM containing 10% FBS); group LIF, in which the cells were treated with complete medium containing 10 ng/ml LIF; group bFGF, in which the cells were treated with complete medium containing 10 ng/ml bFGF; combination group, in which the cells were treated with complete medium containing 10 ng/ml LIF and 10 ng/ml bFGF. The growth curves of hUC-MSC at passage 4 in different groups were assayed by cell counting kit 8. Cellular morphologic changes were observed under inverted phase contrast microscope; hUC-MSC senescence in different groups was detected by β-galactosidase staining. The expression of PCNA, P16, P21, P53, OCT4 and NANOG genes was detected by RT-PCR.</p><p><b>RESULTS</b>The cell growth curves of each group were similar to the S-shape; the cell proliferation rate from high to low as follows: that in the combination group > group bFGF > group LIF > control group. Senescence and declining of proliferation were observed at hUC-MSC very early in control group; the cells in group LIF maintained good cellular morphology at early stage, but cell proliferation was slow and late senescence was observed; a few cells in group bFGF presented signs of senescence, but with quick proliferation; the cells in combination group grew quickly and maintained cellular morphology of hUC-MSC for long time. The LIF and bFGF up-regulated the expression of PCNA, OCT4 and NANOG, while they down-regulated the expression of P16, P21, P53, and their combinative effects were more significant.</p><p><b>CONCLUSION</b>LIF combined with bFGF not only can promote the proliferation and maintenance of stemness of hUC-MSC, but also can delay the senescence of hUC-MSC.</p>