FOXD1 expression-based prognostic model for uveal melanoma.

FOXD1, a new member of the FOX transcription factor family, serves as a mediator and biomarker for cell reprogramming. But its contribution to prognosis of uveal melanoma (UVM) is unclear. This study demonstrated that FOXD1 might promote tumor growth and invasion, because FOXD1 expression was negatively correlated with overall survival, progression-free survival, and disease-specific survival in UVM patients. This conjecture was verified in cell culture with human uveal melanoma cell line (MUM2B) as model cells. Additionally, the biological mechanisms of FOXD1 based on FOXD1-related genomic spectrum, molecular pathways, tumor microenvironment, and drug treatment sensitivity were examined using The Cancer Genome Atlas (TCGA) database, aiming to reasonably explain why FOXD1 leads to poor prognosis of UVM. On these bases, a novel tumor prognostic model was established using the FOXD1-related immunomodulators TMEM173, TNFRSF4, TNFSF13, and ULBP1, which will enable the stratification of disease seriousness and clinical treatment for patients.

An efficient approach for extraction of polysaccharide from abalone (Haliotis Discus Hannai Ino) viscera by natural deep eutectic solvent.

In this study, a natural deep eutectic solvent (NADES) was proposed for the ultrasonic-assisted extraction of polysaccharides from abalone (Haliotis Discus Hannai Ino) viscera. Eleven NADESs were employed for abalone viscera polysaccharide (AVP) extraction. NADES, composed of choline chloride and ethylene glycol in a molar ratio 1: 3 had the highest extraction efficiency. The optimal extraction conditions were obtained using a four-factor, three-level Box-Behnken design and specific response surface methodology. The maximum predicted polysaccharide yield was 17.32 %. Fick's second law was fitted to the extraction process of AVP by ultrasonic-assisted NADES based on a high linear correlation (R2 ≥ 0.9). The extraction rate constants (k), diffusion coefficients (Du) and half-lives (t1/2) were calculated. Compared to the polysaccharides prepared by the conventional method, the polysaccharides extracted by NADES had a higher sugar content, lower molecular weight, more glucuronic acid, and stronger antioxidant capacity. Overall, the NADES extraction method established in this research can become a strategy for the preparation of high-purity and highly bioactive abalone viscera polysaccharides, which has implications for the exploitation and application of marine food byproduct resources.

Detection of membrane protein-protein interaction in planta based on dual-intein-coupled tripartite split-GFP association.

Despite the great interest in identifying protein-protein interactions (PPIs) in biological systems, only a few attempts have been made at large-scale PPI screening in planta. Unlike biochemical assays, bimolecular fluorescence complementation allows visualization of transient and weak PPIs in vivo at subcellular resolution. However, when the non-fluorescent fragments are highly expressed, spontaneous and irreversible self-assembly of the split halves can easily generate false positives. The recently developed tripartite split-GFP system was shown to be a reliable PPI reporter in mammalian and yeast cells. In this study, we adapted this methodology, in combination with the ß-estradiol-inducible expression cassette, for the detection of membrane PPIs in planta. Using a transient expression assay by agroinfiltration of Nicotiana benthamiana leaves, we demonstrate the utility of the tripartite split-GFP association in plant cells and affirm that the tripartite split-GFP system yields no spurious background signal even with abundant fusion proteins readily accessible to the compartments of interaction. By validating a few of the Arabidopsis PPIs, including the membrane PPIs implicated in phosphate homeostasis, we proved the fidelity of this assay for detection of PPIs in various cellular compartments in planta. Moreover, the technique combining the tripartite split-GFP association and dual-intein-mediated cleavage of polyprotein precursor is feasible in stably transformed Arabidopsis plants. Our results provide a proof-of-concept implementation of the tripartite split-GFP system as a potential tool for membrane PPI screens in planta.