Objective To optimize the soaking treatment method for increasing the germination of Amomum villosum Lour.(AVL) seeds.Methods Nine groups were set up: the control group(without soaking treatment),treatment group 1(soaking AVL seeds with naphthylacetic acid 15 mg/mL for 8 hours),treatment group 2(soaking with gibberellin 100 mg/L for 30 hours) and treatment groups 3~8(soaking with clean water for 8,20,30,40,45 and 50 hours respectively).The sowing amount was 100 grains in each group for each time,and the seeds were planted in the outdoor pot at a planting space of 4cm?4cm.Results Naphthylacetic acid had an obvious inhibition on the germination of AVL seeds and delayed the shooting.Gibberellin promoted the germination and shooting of the seeds.Soaking with clean water for 8 hours had no obvious effect on the seed germination,but soaking for 20~50 hours increased the seed germination to various degrees,in particular soaking for 30~50 hours.Conclusion Soaking with gibberellin or clean water can increase the germination of AVL seeds.This method is simple and practical and economic,and is worth of extensively applying in the sowing and breeding of AVL seeds.

Objective:To investigate the effect of serine/arginine-rich splicing factor 2 (SRSF2) on ferroptosis and its possible mechanism in glioblastoma cells.Methods:The online database of gene expression profiling interactive analysis 2 (GEPIA 2) and Chinese Glioma Genome Atlas were used to analyze the expression of SRSF2 in glioblastoma tissue and its association with patients prognosis. To validate the findings of the online databases, the pathological sections of glioblastoma and non-tumor brain tissues from Tianjin Medical University General Hospital, Tianjin, China were collected and analyzed by using immunohistochemistry. Silencing SRSF2 gene expression in glioblastoma cells by siRNA was analyzed with Western blot. The proliferation index was detected by using CCK8 assay. The rescued experiment was conducted by using expression plasmid of pcDNA3.1(+)-SRSF2. The activity of ferroptosis was assessed by using the levels of iron ions and malondialdehyde in glioblastoma cells and the changes in the ratio of glutathione to oxidized glutathione. The changes of gene expression and differential pre-mRNA alternative splicing (PMAS) induced by SRSF2 were monitored by using the third-generation sequencing technology analysis, namely Oxford nanopore technologies (ONT) sequencing analysis.Results:SRSF2 expression was higher in glioblastoma tissues than non-tumor brain tissues. Immunohistochemistry also showed a positive rate of 88.48%±4.60% in glioblastoma tissue which was much higher than the 9.97%±4.57% in non-tumor brain tissue. The expression of SRSF2 was inversely correlated with overall and disease-free disease survivals ( P<0.01). The proliferation index of glioblastoma cells was significantly reduced by silencing with SRSF2 siRNA ( P<0.01) and could be reversed with transfection of exogenous SRSF2. The levels of intracellulariron ions and malondialdehyde increased ( P<0.05), but the glutathione/oxidized glutathione ratio and the expression of key proteins in the glutathione pathway remained unchanged ( P>0.05). ONT sequencing results showed that silencing SRSF2 in glioblastoma cells could induce a significant alternative 3' splice site change on ferroptosis suppressor protein 1 (FSP1). Conclusion:SRSF2 inhibits the ferroptosis in glioblastoma cells and promotes their proliferation, which may be achieved by regulating FSP1 PMAS.