Mechanistic analysis for the origin of diverse diterpenes in Tripterygium wilfordii

Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3-23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.

Effects of subthreshold micropulse yellow laser (577 nm) on VEGF, NGF and chemerin expressions in retina of early stage diabetic rats / 眼科新进展

Objective To investigate the effects of subthreshold micropulse yellow laser (577 nm) on vascular endothelial growth factor (VEGF),nerve growth factor (NGF) and Chemerin expressions in retina of early stage diabetic rats.Methods A total of 40 Brown Norway rats were treated with streptozocin (65 mg · kg-1) to establish the diabetic model.20 diabetic BN rats' right eyes were received subthreshold micropulse yellow laser (577 run) therapy after 2 weeks.The left eyes were used as control group.At 3 days,7 days,14 days,28 days after laser therapy,5 BN rats were randomly chosen to perform RT-PCR and Weston-blot.The expressions of mRNA and protein of VEGF,NGF and Chemerin were analyzed.Results The expression of VEGF mRNA and protein increased in control group at 3 days,7 days,14 days and 28 days (all P ＜ 0.05).Compared with the control group,VEGF mRNA and protein decreased in the subthreshold micropulse yellow laser (577 nm) group (all P ＜ 0.05).The expression of NGF mRNA and protein decreased in the control group at 3 days,7 days,14 days and 28 days (all P ＜ 0.05),however,the difference was not statistically significant between 3 days and 7 days(P ＞0.05).Compared with control group,NGF mRNA and protein increased in the subthreshold micropulse yellow laser (577 nm) group (all P ＜ 0.05),with maximum expression at 14 days.The expression of chemerin mRNA and protein increased at 3 days,7 days,14 days and 28 days in the control group (all P ＜0.05).Compared with the control group,chemerin mRNA and protein decreased in the subthreshold micropulse yellow laser (577 urn) group (all P ＜ 0.05).Conclusion Subthreshold micropulse yellow laser (577 urn) can suppress VEGF,Chemerin expression and upregulate NGF expression in early stage diabetic rats.

Effect of leptin on the oxidative damage in human retinal pigment epithelial cells / 中华眼底病杂志

Objective To investigate the effects of leptin on the oxidative damage in human retinal pigment epithelial (RPE) cells.Methods Human RPE cells (ARPE-19) were cultured in vitro,and randomly divided into control group and insulin resistance group.RPE cells were treated with 0,10,100 ng/mL leptin for 24,48,72 hours respectively.Then the levels of reactive oxygen species (ROS) expression in RPE cells were detected by 2',7'-dichlorofluorescin-diacetate (DCFH-DA),and the levels of 8-hydroxy-2′-deoxyguanosine (8-OHdG) expression in RPE cells were observed by immunocytochemistry (ICC),and the levels of human 8-oxoguanine DNA glycosylase 1 (hOGG1) expression in lysate were measured by Western blot.Results After 24,48,72 hours,the level of ROS (Control group:F=37.136,37.178,49.634; P＜0.05.Insulin resistance group:F=9.822,28.881,71.150;P＜0.05),8-OHdG (Control group:F =88.643,390.920,1039.276; P ＜ 0.05.Insulin resistance group:F =273.311,299.155,82.237;P＜0.05) and hOGGl (Control group:F=470.062,1073.113,295.456;P＜0.05.Insulin resistance group:F =240.032,592.389,527.760 ; P＜0.05) expression increased significantly with the increase of leptin concentration in control group and insulin resistance group.Under the same leptin concentration,the level of 8-OHdG has a trend that it was higher in the insulin resistance group than the control group.After 24 hours,the difference of hOGGl expression between control group and insulin resistance group was not significant (F=23.392,P＞0.05).After 72 hours,the level of hOGGl expression was significantly higher in the insulin resistance group than the control group (F=129.394,P＜0.05).The level of hOGGl expression was significantly higher at 48 hours than that at 24 hours and 72 hours (P＜ 0.05).Conclusion Leptin could induce the oxidative damage of RPE cells in normal and insulin resistance status.With the increase of leptin concentration and time extended,the degree of oxidative damage and its repair were both increased.The degree of oxidative repair increased with the increase of leptin concentration,but decreased with time extended.

Research on relationship of the procrastination and the big five personality factors in university students / 中华行为医学与脑科学杂志

Objective To study the relationship of the university students' procrastination and big five personality factors.Methods 461 university students were assessed by the simple edition of tuckman procrastination scale(TPS) and the big five personality inventory short form(NEO-FFI).Results TPS score of the student was (48.51 ± 7.66).There were 89.4% students respectively had different level of procrastination behavior.There were no significant differences in university students by the gender(47.95 ±7.84 vs 48.95 ± 7.43 ) ,whether one-child students(48.64 ± 7.49 vs 48.45 ± 7.81 ) and grades.There were significant positive correlation between procrastination and the personality factor of neuroticism( r=0.235, P＜0.01 ) and significant negative correlation with extraversion ( r=-0.180, P ＜ 0.01 ) and conscientiousness ( r =-0.198, P ＜ 0.01 ).The big five personality factors of neuroticism,extraversion and conscientiousness could explain the 14.0% variation of procrastination of university students.Conclusion Procrastination is a very common phenomenon in the university students.The students with higher neuroticism, lower extraversion and conscientiousness of big five personality have more procrastination behavior.