Doxycycline Suppresses Vasculogenic Mimicry in Human Pterygium Fibroblasts.

PURPOSE: To explore the effect of doxycycline on vasculogenic mimicry (VM) formation and the potential mechanism in human pterygium fibroblasts in order to find novel targets for pterygium therapy. METHODS: First, we demonstrate the existence of VM in 73 pterygium specimens by CD31 and periodic acid Schiff (PAS) dual staining. Then we used cell counting kit-8, clone formation assay and flow cytometry to prove the inhibitory effect of doxycycline on cell proliferation and apoptosis. The VM formation was evaluated through wound healing assay, cell transwell assay and three-dimensional cell culture combined with PAS staining. Finally, we used Western blot to testify the correlation of the VM and the factors in protein level preliminarily. RESULTS: Our results showed that VM existed in human pterygium specimens exactly. Otherwise, in human pterygium fibroblasts, doxycycline induced a dose-dependent inhibitory effect on cell proliferation and apoptosis induction. Besides, doxycycline significantly suppressed vasculogenic mimicry tube formation, cell migration and invasion. Furthermore, doxycycline impaired the expression of MMP-9, MMP-2 and VEGF which may related to pterygium VM formation. CONCLUSIONS: Doxycycline decelerated pterygium progression might be through inhibiting VM formation according to the downregulation of MMP-9, MMP-2 and VEGF, which may provide the basis of further studies involving doxycycline for pterygium treatment.

[Optimization of application parameters of soil seed bank in vegetation recovery via response surface methodology].

The thickness of surface soil, the covering thickness and the number of adding arbor seeds are all important factors to be considered in the application of soil seed bank (SSB) for vegetation recovery. To determine the optimal conditions, the Box-Behnken central composite design with three parameters and three levels was conducted and Design-Expert was used for response surface optimization. Finally, the optimal model and optimal level of each parameter were selected. The quadratic model was more suitable for response surface optimization (P < 0.0001), indicating the model had good statistical significance which could express ideal relations between all the independent variable and dependent variable. For the optimum condition, the thickness of surface soil was 4.3 cm, the covering thickness was 2 cm, and the number of adding arbor seeds was 224 ind x m(-2), under which the number of germinated seedlings could be reached up to 6222 plants x m(-2). During the process of seed germination, significant interactions between the thickness of surface soil and the covering thickness, as well as the thickness of surface soil and the number of adding arbor seeds were found, but the relationship between the covering thickness and the number of adding arbor seeds was relatively unremarkable. Among all the parameters, the thickness of surface soil was the most important one, which had the steepest curve and the largest standardized coefficient.