Objective This study aims to achieve high-yield functional expression of the human voltage-gated potassium channel K_V3.1 using an Escherichia coli cell-free protein synthesis system, thereby providing a novel synthetic approach for drug screening, structural analysis and functional characterization of K_V3.1. Methods K_V3.1 was expressed in an Escherichia coli cell-free protein synthesis system for 10 hours in the presence of peptide surfactant A₆K. The secondary structure of K_V3.1 was analyzed by circular dichroism spectroscopy. The potassium channel activity of the recombinant protein liposome K_V3.1-A₆K was investigated using fluorescent dyes Oxonol VI as indicators, which are capable of reflecting alterations in membrane potential. Results Soluble K_V3.1 protein was successfully synthesized, achieving a purified yield of up to 1.2 mg/mL via an Escherichia coli cell-free protein synthesis system. Circular dichroism spectroscopy revealed that K_V3.1 exhibited characteristic α-helical secondary structures. Membrane potential fluorescence assays demonstrated that the K_V3.1-A₆K proteoliposomes, which were reconstructed with surfactant peptide A₆K, exhibited remarkable potassium ion permeability. Conclusion This study successfully achieved high-yield expression of human K_V3.1 with activity using an Escherichia coli-based cell-free protein synthesis system. This innovative method not only significantly enhances the expression yield of K_V3.1, but also maintains its functional activity, thereby establishing a novel and efficient synthetic platform for drug screening and advancing our understanding of structure-function relationships in K_V3.1 research.
Humans ; Escherichia coli/metabolism* ; Shaw Potassium Channels/biosynthesis* ; Cell-Free System ; Circular Dichroism ; Protein Biosynthesis ; Recombinant Proteins/metabolism* ; Membrane Potentials ; Shab Potassium Channels

Organ damage from cancer treatment remarkably effects patients’ prognosis and quality of life. In recent years, preventive organ protection strategies, such as interdisciplinary collaboration, early prevention, precision interventions, psychological support, and the integrated application of traditional Chinese medicine, have demonstrated substantial clinical value and achieved notable progress. However, these approaches still encounter multiple challenges. Establishing multidisciplinary teams, optimizing therapeutic balance, and strengthening evidence-based research are essential for addressing the challenges related to treatment balance optimization, multidisciplinary coordination, and clinical translation of novel technologies. This review systematically summarizes recent advancements in preventive organ protection, analyzes existing challenges and potential solutions, and offers forward-looking recommendations. It aims to provide valuable insights for optimizing comprehensive cancer treatment strategies and improving long-term patient outcomes.

Abstract Dry eye disease is a chronic ocular surface inflammatory disease caused by abnormal tear quality or quantity and decreased tear film stability due to various reasons, and often accompanied by ocular discomfort such as itching, dryness, foreign body sensation, and visual dysfunction, which can seriously affect the patient′s quality of life and visual quality if not intervened in time. With the change of social lifestyles, the increase of environmental pollution and the trend of population aging, dry eye disease has become the most common ocular surface disease besides refractive error. Currently, dry eye disease is widely recognized as a non-infectious immune-related inflammatory disease, but the signaling pathways involved in dry eye disease are poorly understood. Whether dry eye disease is caused by excessive tear evaporation, insufficient tear production, or mucin deficiency, the ocular surface tissues(cornea/conjunctiva) inevitably undergo pathological processes such as aberrant proliferation, squamous epithelial hyperplasia, initiation of corneal damage repair mechanisms, and reduction in the number of conjunctival goblet cells, whereas the Wnt/β-catenin pathway is known to have a wide range of biological functions and plays an important role in cell proliferation, differentiation, and stemness maintenance. Therefore, this review describes the pathogenesis and potential experimental therapeutic options of the Wnt/β-catenin signaling pathway in dry eye disease from this perspective, aiming to provide new targets for the treatment of dry eye disease and achieve the goal of controlling the disease progression from the root.

Objective:Using atenolol as a model drug,the aim of this study was to develop a sustained and controlled transdermal drug delivery system(TDDS)based on polyethyleneimine-modified MoS2 nanoparticles(PEI-MoS2 NPs)that were responsive to near infrared(NIR)laser irradiation.Methods:The three-dimensional flower-like PEI-MoS2 NPs were successfully synthesized and further characterized by attenuated total reflection Fourier transform infrared spectroscopy,X-ray diffraction measurements,scanning electron microscopy,and transmission electron microscopy.The controlled release capacity of PEI-MoS2 NPs was examined using in vitro drug release and skin penetration experiments.Results:The PEI-MoS2 NPs exhibited a drug loading efficiency of 53.86% and high photothermal conversion ability.Moreover,the release of atenolol was enhanced by NIR stimulation with an enhancement ratio of 1.56.Conclusion:NIR-controlled PEI-MoS2 NPs was essential for the control and sustained release of drugs in TDDS.

AIM:To explore the effects of CD38 on lysosome reformation and cholesterol efflux in macro-phages.METHODS:Bone marrow-derived macrophages from low-density lipoprotein(LDL)receptor knockout(LDLr-/-)mice were cultured as cell model.Live cell imaging system was applied to evaluate the effect of nicotinic acid adenine di-nucleotide phosphate(NAADP)on lysosome number.ELISA was conducted to measure NAADP level in macrophages.After the cells were treated with nicotinic acid(NA),RT-qPCR was conducted to detect CD38 mRNA expression,and Western blot was conducted to observe CD38 protein expression and phosphorylated transcription factor EB(TFEB)level.Laser scanning confocal microscopy was applied to evaluate the influence of CD38/NAADP signaling on lysosome number and cholesterol egression.RESULTS:NAADP remarkably increased lysosome number(P<0.05),and this effect was significantly inhibited by NAADP antagonist NED-19,Ca2+ chelator BAPTA,and calcineurin inhibitor CsA(P<0.05).CD38 markedly enhanced NAADP synthesis in macrophages(P<0.05).NAADP synthetic substrate NA prominently ele-vated the expression of CD38 mRNA and protein(P<0.05).NA significantly decreased the phosphorylated TFEB level;this effect was also attenuated by NED-19,BAPTA and CsA(P<0.05).Disrupting CD38/NAADP signaling pathway markedly inhibited NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux in macrophages(P<0.05).NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux abolished in LDLr/CD38 DKO macrophages(P<0.05),whereas these effects induced by NA were recovered after CD38 gene rescue.CONCLUSION:CD38 triggers lysosome reformation via TFEB and consequently pro-motes the efflux of lysosomal free cholesterol and cytosol cholesterol ester.

The ultrasonic manifestations of benign and malignant breast imaging-reporting and data system(BI-RADS)4 lesions overlap in some degrees,is able to result in unnecessary biopsy or untimely therapy.Accurate classifying the nature of BI-RADS 4 breast lesions can provide reliable references for clinical decision-making.The progresses of application of new ultrasonic technologies,including automated breast volume scanner,superb micro-vascular imaging,elastography,contrast-enhanced ultrasound and artificial intelligence for assisting diagnosis of BI-RADS 4 lesions were reviewed in this article.

Zedoary turmeric oil， volatile oil extracted from zedoary turmeric， composed mainly of monoterpenes （including α-pinene， β-pinene， etc.） and sesquiterpenes （including β-elemene， zedoary alcohol， zedoary ketone， etc.）， and has been used in clinical practice to treat various malignant tumors such as ovarian cancer， cervical carcinoma， colorectal cancer， lung cancer and liver cancer. Zedoary turmeric oil regulates vascular endothelial growth factor and nuclear factors- κB， signal transducers and activator of transcription 3 signaling pathways to play a role in inhibiting tumor angiogenesis， inhibiting tumor cell proliferation， inducing tumor cell apoptosis， and blocking cell cycle. However， due to its insolubility in water and poor stability， its clinical application is limited； the application of new formulations and technologies such as liposomes， microspheres， and nanoemulsion improves the solubility and stability of zedoary turmeric oil. This paper summarizes recent research progress on the chemical composition， anti-tumor effects， and formulations of zedoary turmeric oil， both domestically and internationally， providing a reference for further expanding the clinical application and formulation development of zedoary turmeric oil in the anti-tumor field.

Zedoary turmeric oil， volatile oil extracted from zedoary turmeric， composed mainly of monoterpenes （including α-pinene， β-pinene， etc.） and sesquiterpenes （including β-elemene， zedoary alcohol， zedoary ketone， etc.）， and has been used in clinical practice to treat various malignant tumors such as ovarian cancer， cervical carcinoma， colorectal cancer， lung cancer and liver cancer. Zedoary turmeric oil regulates vascular endothelial growth factor and nuclear factors- κB， signal transducers and activator of transcription 3 signaling pathways to play a role in inhibiting tumor angiogenesis， inhibiting tumor cell proliferation， inducing tumor cell apoptosis， and blocking cell cycle. However， due to its insolubility in water and poor stability， its clinical application is limited； the application of new formulations and technologies such as liposomes， microspheres， and nanoemulsion improves the solubility and stability of zedoary turmeric oil. This paper summarizes recent research progress on the chemical composition， anti-tumor effects， and formulations of zedoary turmeric oil， both domestically and internationally， providing a reference for further expanding the clinical application and formulation development of zedoary turmeric oil in the anti-tumor field.

As a severe clinical manifestation of nonalcoholic fatty liver disease， nonalcoholic steatohepatitis （NASH） is characterized by lipid deposition and inflammatory damage in the liver. At present， clinical medications for NASH are still in the exploratory phase， and it is urgent to make progress. Recent studies have shown that the pathogenesis of NASH is associated with circadian rhythm disorders in the liver， with the specific manifestation of dysregulated expression of liver clock genes such as BMAL1， which increases hepatic lipogenesis， reduces fatty acid oxidation， and activates pro-inflammatory factors. Therefore， improving circadian rhythm of the liver and regulating the expression of liver clock genes are feasible strategies for the prevention and treatment of NASH. Currently， some medications for NASH via activating the proteins encoded by clock genes have been applied in animal experiments， for example， the REVERB full-agonist SR9009 can inhibit the development of liver inflammation， which confirms the possibility of NASH treatment by targeting the proteins encoded by clock genes. This article summarizes the role of hepatic clock genes in regulating lipid metabolism and the development and progression of inflammation in the liver and elaborates on the recent advances in medications targeting clock genes and the proteins encoded by clock genes， in order to provide new targets for the treatment of NASH.