Objective:To identify the role of transcription factors Sp1 and Sp3 in the expressional regulation of ezrin in human esophageal carcinoma cells.Methods:Esophageal carcinoma EC109 cells were transfected with expressing vectors CMV-Sp1 or CMV-Sp3,and the effect of Sp1 and Sp3 over-expression on ezrin mRNA and protein expression was determined by real time RT-PCR and Western blot analysis.Furthermore,EC109 cells were cotransfected with the ezrin promoter-directed luciferase reporter vector and control vector pRL-TK along with transcription factor expression vector.The roles of Sp1 and Sp3 in ezrin promoter activation and whether this activation occurred through the Sp1 binding site,-75/-69,were analyzed by dual-luciferase reporter assay system.Results:Over-expression of transcription factors Sp1 and Sp3 significantly increased the expression of ezrin mRNA and protein and the ezrin promoter activity in EC109 cells.Sp1 and Sp3 enhanced the promoter activity through different binding sites and only Sp1 did that through the-75/-69 site.Conclusion:Sp1 and Sp3 can regulate ezrin expression in EC109 cells.

Previous studies suggest that NGAL (neutro phil gelatinase-associated lipocalin) is involved in the transformation and development of esophageal carcinoma. Alteration of NGAL expression can trigger the change of cellular morphology in esophageal carcinoma cells. However, the mechanisms remain unclear. To get a better understanding of NGAL function in esophageal carcinoma, NGAL protein was expressed in methylotrophic yeast, Pichia pastoris, and purified by chromatography. EC1.71 cells expressed high levels of NGALR (NGAL receptor) and EC109 cells expressed low levels of NGALR were used as cells model. The trafficking and the possible function of NGAL protein were then analyzed in the esophageal carcinoma cells. The results showed that 5-FAM-labeled recombinant NGAL protein could internalize into the EC1.71 and EC109 cells. Furthermore, the internalized NGAL protein could induce the alteration of cellular morphology, resulting in generation of autophagosome, transcriptional up-regulation of genes associated with autophagy and increase of phospho-ERK1/2 (p-ERK1/2). Interestingly, the treatment with the NGAL protein did not affect the intracellular iron level. These data indicate that induced autophagy by exogenous NGAL protein is a mechanism that internalized NGAL plays important roles in esophageal carcinoma cells, independent with NGAL-mediated iron transport process, while ERK1/2 signal pathway is involved in activation of autophagy by exogenous NGAL protein.

Insomnia is the most common sleep disorder in clinical practice， and prolonged insomnia can significantly impact daily life and work. Currently， the clinical treatment of insomnia primarily relies on sedative and hypnotic drugs， which， although fast-acting， come with numerous adverse reactions. Traditional Chinese medicine （TCM） approaches the treatment of insomnia by employing syndrome differentiation and focusing on addressing the root cause of the disease. This approach is characterized by fewer adverse reactions， stable long-term effects， and better patient compliance. Huanglian Ejiaotang was first recorded in Zhang Zhongjing's Treatise on Febrile and Miscellaneous Diseases（《伤寒杂病论》） during the Han dynasty. It has the effects of nourishing Yin， reducing fire， eliminating irritability， and calming the nerves. Clinical studies have shown that Huanglian Ejiaotang is frequently used to treat insomnia and， when combined with other classic prescriptions， Western medicine， or other therapies， it can have a synergistic effect， thereby enhancing therapeutic outcomes. Research indicates that the mechanism of action of Huanglian Ejiaotang in treating insomnia may be related to the regulation of the hypothalamic-pituitary-adrenal （HPA） axis， 5-hydroxytryptamine （5-HT）， norepinephrine （NE）， dopamine （DA）， central neurotransmitters， gut microbiota， and the inhibition of inflammatory factors. The active components of Huanglian Ejiaotang that are effective in treating insomnia include berberine， baicalin， baicalein， total glucosides of paeony， and collagen. This article discussed the treatment of insomnia with Huanglian Ejiaotang from several perspectives， including its theoretical foundation， clinical research， and mechanisms of action， and also summarized the latest research progress on Huanglian Ejiaotang for insomnia， with the aim of providing more comprehensive ideas for the clinical diagnosis and treatment of insomnia.