Hsa_circ_0087354 Regulates Proliferation and Redox State via hsa-miR-199-3p / SLC7A11 in MG-63 Cells / 中国生物化学与分子生物学报

Circular RNAs (circRNAs),a kind of novel non-coding RNAs, have been shown to play animportant role in cellular redox reactions. In the previous study, we found that hsa_circ_0087354 wasclosely related to the cellular redox state by real-time PCR. After overexpression of hsa_circ_0087354, the relative expression of ROS1 were decreased significantly (P < 0. 01), while the levels of SOD1 wereincreased significantly (P < 0. 05) . The activities of SOD and Gpx as well as GSH concentration weresignificantly increased (P < 0. 01), and cell proliferation was promoted in cells (P < 0. 05) . Bioinformatics analysis predicted that there were binding sites between hsa-miR-199-3p and hsa_circ_0087354 as well as solute carrier family 7 member 11 (SLC7A11), which might have a targetedregulatory relationship. Dual luciferase reporter assay confirmed the targeted regulatory relationshipbetween hsa-miR-199-3p with hsa_circ_0087354, and SLC7A11. Overexpressed hsa_circ_0087354 plasmid and ctrl plasmid were constructed, hsa-miR-199a-3p, hsa-miR-199b-3p and hsa-miR-NC mimicswere synthesized. Real-time PCR analysis showed that the relative expression of hsa-miR-199-3p was observably decreased (P < 0.01), while the relative expression of SLC7A11 in cells was dramaticallyincreased after transfection of has_circ_0087354 plasmid (P < 0.05) . After transfection with hsa-miR-199-3p, the relative expressions of SLC7A11 were markedly decreased (P < 0.001) . The activities ofSOD and Gpx, GSH concentration (P<0.01), and cell proliferation rate (P < 0.05) were significantlyreduced. In conclusion, hsa_circ_0087354 could enhance the expression of SLC7A11, promote theproliferation of cells and reduce the oxidative stress by adsorption of hsa-miR-199-3p.

Quantitative Analysis for Telomerase Activity in Rice / 中国生物工程杂志

The Telomeric Repeat Amplification Protocol(TRAP)and its modified versions change the size and/or the ratio of the telomerase products in the amplification stage of the assay.Based on TRAP,a useful method was developed for detecting telomerase activity in rice.A special precursor primer and a special reverse primer and conducted two steps of PCR cycles were designed.GENE Genius? Bio-imaging System was applied for this quantitative analysis for exploring telomerase activity and its optimal reaction conditions.The method ensured that the optimal reaction conditions for the telomerase was 19℃,13minutes,at a concentration of 0.28 u g/? l.A quantitative analysis method was established for detecting telomerase activity in rice.With this method,we detected telomerase activity in roots,young leaves and young panicles of six parental lines of hybrid rice.The results show that young panicles have the highest telomerase activity,demonstrating that telomerase activity is closely related to the cell vitality in plants.

Prokaryotic Expression and Characteristics Reseach of Pinellia Ternate and Pinellia Pedatisecta Lectins / 中国生物工程杂志

Pinellia ternate and Pinellia pedatisecta lectins with biological functions could be obtained through procaryon expression.Their biological similarities and differences were analyzed and further experiments were conducted to discuss the agglutinative activity difference between polymer lectin and metamer lectin.The results showed that agglutinative activity of Pinellia pedatisecta was four times more than that of Pinellia ternate.The differences of the agglutinative activity and pharmacologic action were mainly because two site mutations of Pinellia pedatisecta in the third active site compared with the amino acids sequence in Pinellia ternate.In addition,the metamer lectin expressed by prokaryotic expression system did not agglutinate rabbit red blood cells.It conduct further discussion about the difference between two lectins and the difference between polymer lectin and metamer lectin.It would be applicable for solving the problem of lacking Pinellia resources.