ABSTRACT
Objective:To explore the effect of knockdown of the homeobox gene paired-box 6 ( Pax6) on the biological behavior and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs). Methods:The SRA01/04 human LECs were divided into small interfering RNA-Pax6 (siRNA-Pax6) group transfected with siRNA-Pax6 and siRNA negative control (siRNA-NC) group transfected with disordered siRNA.Cell survival rate was detected by cell counting kit-8 method at 24, 48 and 72 hours after transfection.Cell cycle distribution and apoptosis were analyzed by flow cytometry at 48 hours after transfection.Migratory capability of cells was examined by cell scratch test at 24 hours after transfection.The mRNA relative expression levels of Pax6, α-crystallin A (CRYAA), α-crystallin B (CRYAB), Sox2, α-smooth muscle actin (α-SMA) and E-cadherin were detected by quantitative real-time PCR at 48 hours after transfection.The relative expression of Pax6 protein was detected by Western blot at 48 hours after transfection.Results:There was a significant difference in cell survival rates at different time points between the two groups ( Fgroup=4.776, P<0.05; Ftime=13.535, P<0.05). The cell survival rate of siRNA-Pax6 group was obviously lower than that of siRNA-NC group at 48 and 72 hours after transfection, and the differences were statistically significant (both at P<0.05). Compared with siRNA-NC group, the proportion of cells in G 0/G 1 phase was significantly increased and the proportion of cells in S phase was significantly reduced in siRNA-Pax6 group ( t=9.971, -5.063; both at P<0.05). The cell migration rate of siRNA-Pax6 group was (19.73±6.07)%, which was lower than (70.56±2.97)% of siRNA-NC group, showing a statistically significant difference ( t=-7.245, P<0.05). The relative expressions of Sox2 mRNA and α-SMA mRNA were lower, and the relative expression of E-cadherin mRNA was higher in siRNA-Pax6 group than siRNA-NC group, with statistically significant differences between them ( t=-23.254, -5.294, 6.062; all at P<0.01). The relative expression of CRYAA mRNA and CRYAB mRNA was significantly higher in siRNA-Pax6 group than siRNA-NC group, and the differences were statistically significant ( t=5.521, 8.270; both at P<0.01). The relative expressions of Pax6 mRNA and protein in siRNA-Pax6 group were 0.27±0.01 and 0.24±0.05, respectively, which were both lower than 1.00±0.05 and 1.14±0.10 in siRNA-NC group, showing statistically significant differences ( t=-14.456, -4.458; both at P<0.001). Conclusions:Silence of Pax6 can suppress the proliferation and EMT of human LECs and enhance the expression of crystallin.
ABSTRACT
Objective To explore the teaching effects of theory and practice integrated teaching model in small class teaching of ENT nursing. Methods Two classes of nursing students from grade two were randomly selected as experiment group and control group with 120 students in each group. The experiment group was treated with theory and practice integrated teaching model in small class and the control group with traditional practice. The teaching effects of the two groups were compared by test results as well as questionnaire survey . Result The teaching effect in the experiment group was significantly better than that of the control group (P<0.05). Conclusion The theory and practice integrated teaching model in small classes can promote the students' interest in learning and mobilize their initiative and enthusiasm to master theoretical knowledge and specialist skills.
ABSTRACT
Objective:To examine the expression and function of thrombonspondin-1 (TSP-1) in oxygen-induced retinopathy in new-born mice, and to investigate its role in retinal neovascularization. Methods:A total of 40 C57BL/6J newborn mice were divided equally into a model group (n=20)andanormalcontrolgroup(n=20).Miceinthemodelgroupwereexposedto75%oxygentoestablish the oxygen-induced retinopathy (OIR) model. On the 7th, 9th, and 11th day after the birth of mice, 5 mice were randomly selected each time from the 2 groups to examine the expression of TSP-1 mRNA with reverse transcription polymerase chain reaction (RT-PCR). After that, 5 mice were selected on the 11th day to observe the retinal neovascularization by fluorescein angiography retinal flatmount. Results:On the 11th day, fluorescein angiography retinal flatmount showed that the retinal blood vessels presented mean network distribution in the normal control group, while in the model group, a lot of dilatated areas in the retinal main vessels surrounded the optic disc. Meanwhile lots of new blood vessels were found surrounding the optic disc with irregular distribution but well distributed peripherial retinal small vessels, which was typical of early stage OIR. There was no signiifcant difference in the retinal TSP-1 mRNA level between the model group and the normal control group in the postnatal 7-day mice (P>0.05). Compared with the normal control group, the expression of TSP-1 mRNA in the model group was signiifcantly lower in postnatal 9-day and 11-day mice (P Conclusion:In the early stage of OIR model (retinal vascular growth and development stage), the expression of TSP-1 mRNA in the retinal tissue is gradually decreased, implying that TSP-1 (as a negative regulatory factor) may be involved in the formation of retinal neovascularization in the early stage.