To develop a new high-throughput screening model for human high-density lipoprotein(HDL)receptor(CD36 and LIMPⅡanalogous-1,CLA-1)agonists using CLA-1-expressing insect cells.Methods With the total RNA of human hepato...To develop a new high-throughput screening model for human high-density lipoprotein(HDL)receptor(CD36 and LIMPⅡanalogous-1,CLA-1)agonists using CLA-1-expressing insect cells.Methods With the total RNA of human hepatoma cells BEL-7402 as template,the complementary DNA(cDNA)of CLA-1 was amplified by reverse transcription-polymerase chain reaction(RT-PCR).Bac-to-Bac baculovirus expression system was used to express CLA-1 in insect cells.CLA-1 cDNA was cloned downstream of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus(AcNPV)into donor vector pFastBacl and recombinant pFastBacl-CLA-1 was transformed into E.coli DH10Bac to transpose CLA-1 cDNA to bacrnid DNA.Recombinant bacrnid-CLA-1 was transfected into Spodopterafrugiperda Sf9 insect cells to produce recombinant baculovirus particles.Recombinant CLA-1 was expressed on the membrane of Sf9 cells infected with the recombinant baculoviruses.A series of parameters of DiI-lipoprotein binding assays of CLA-1-expressing Sf9 cells in 96-well plates were optimized.Results Western blot analysis and DiI-lipoprotein binding assays confirmed that CLA-1 expressed in insect cells had similar immunoreactivity and ligand binding activity as its native counterpart.A reliable and sensitive in vitro cell-based assay was established to assess the activity of CLA-1 and used to screen agonists from different sample libraries.Conclusion Human HDL receptor CLA-1 was successfully expressed in Sf9 insect cells and a novel high-throughput screening model for CLA-1 agonists was developed.Utilization of this model allows us to identify potent and selective CLA-1 agonists which might possibly be used as therapeutics for atherosclerosis.展开更多
The high porosity and tunable chemical functionality of metal-organic frameworks(MOFs)make it a promising catalyst design platform.High-throughput screening of catalytic performance is feasible since the large MOF str...The high porosity and tunable chemical functionality of metal-organic frameworks(MOFs)make it a promising catalyst design platform.High-throughput screening of catalytic performance is feasible since the large MOF structure database is available.In this study,we report a machine learning model for high-throughput screening of MOF catalysts for the CO_(2) cycloaddition reaction.The descriptors for model training were judiciously chosen according to the reaction mechanism,which leads to high accuracy up to 97%for the 75%quantile of the training set as the classification criterion.The feature contribution was further evaluated with SHAP and PDP analysis to provide a certain physical understanding.12,415 hypothetical MOF structures and 100 reported MOFs were evaluated under 100℃ and 1 bar within one day using the model,and 239 potentially efficient catalysts were discovered.Among them,MOF-76(Y)achieved the top performance experimentally among reported MOFs,in good agreement with the prediction.展开更多
Lithium-ion batteries(LiBs)with high energy density have gained significant popularity in smart grids and portable electronics.LiMn_(1-x)Fe_(x)PO_(4)(LMFP)is considered a leading candidate for the cathode,with the pot...Lithium-ion batteries(LiBs)with high energy density have gained significant popularity in smart grids and portable electronics.LiMn_(1-x)Fe_(x)PO_(4)(LMFP)is considered a leading candidate for the cathode,with the potential to combine the low cost of Li Fe PO_(4)(LFP)with the high theoretical energy density of LiMnPO_(4)(LMP).However,quantitative investigation of the intricate coupling between the Fe/Mn ratio and the resulting energy density is challenging due to the parametric complexity.It is crucial to develop a universal approach for the rapid construction of multi-parameter mapping.In this work,we propose an active learning-guided high-throughput workflow for quantitatively predicting the Fe/Mn ratio and the energy density mapping of LMFP.An optimal composition(LiMn_(0.66)Fe_(0.34)PO_(4))was effectively screened from 81 cathode materials via only 5 samples.Model-guided electrochemical analysis revealed a nonlinear relationship between the Fe/Mn ratio and electrochemical properties,including ion mobility and impedance,elucidating the quantitative chemical composition-energy density map of LMFP.The results demonstrated the efficacy of the method in high-throughput screening of LiBs cathode materials.展开更多
Additives are widely employed to regulate the morphology,size,and agglomeration degree of crystalline materials during crystallization to enhance their functional,physical,and powder properties.However,the existing me...Additives are widely employed to regulate the morphology,size,and agglomeration degree of crystalline materials during crystallization to enhance their functional,physical,and powder properties.However,the existing methods for screening and validating target additives require a large quantity of materials and involve tedious molecular simulation/crystallization experiments,making them time-consuming,resource-intensive,and reliant on the operator’s experience level.To overcome these challenges,we proposed a computer vision-assisted high-throughput additive screening system(CV-HTPASS)which comprises a high-throughput additive screening device,in situ imaging equipment,and an artificial intelligence(AI)-assisted image-analysis algorithm.Using the CV-HTPASS,we performed high-throughput screening experiments on additives to regulate the succinic acid crystal properties,generating thousands of crystal images with diverse crystal morphologies.To extract valuable crystal information from the massive data and improve the analysis accuracy and efficiency,the AI-based image-analysis algorithm was implemented innovatively for the segmentation,classification,and data mining of crystals with four morphologies to further screen the target additive.Subsequently,scale-up crystallization experiments conducted under optimized conditions demonstrated that succinic acid products exhibited a preferred cubic morphology,reduced agglomeration degree,narrowed crystal size distribution,and improved powder properties.The proposed CV-HTPASS offers a highly efficient approach for scale-up experiments.Further,it provides a platform for the screening of additives and the optimization of the powder properties of crystal products in industrial-scale crystallization processes.展开更多
The capture of CO_(2)from CO_(2)/H_(2)gas mixtures in syngas is a crucial issue for hydrogen production from steam methane reforming in industry,as the presence of CO_(2)directly affects the purity of H_(2).A combinat...The capture of CO_(2)from CO_(2)/H_(2)gas mixtures in syngas is a crucial issue for hydrogen production from steam methane reforming in industry,as the presence of CO_(2)directly affects the purity of H_(2).A combination of a high-throughput screening method and grand canonical Monte Carlo simulation was utilized to evaluate and screen 1725 metal–organic frameworks(MOFs)in detail as a means of determining their adsorption performance for CO_(2)/H_(2)gas mixtures.The adsorption and separation performance of double-linker MOFs was comprehensively evaluated using eight evaluation indicators,namely,the largest cavity diameter,accessible surface area,pore occupied accessible volume,porosity,adsorption selectivity,working capacity,adsorbent performance score and percent regeneration.Six optimal performance frameworks were screened to further study their single-component adsorption and binary competitive adsorption of CO_(2)/H_(2)respectively.The CO_(2)adsorption selectivity at different CO_(2)/H_(2)feed ratios was also evaluated,which indicated their excellent adsorption and separation performance.The microscopic adsorption mechanisms for CO_(2)and H_(2)at the molecular level were investigated by analyzing the radial distribution function and density distribution.This study may provide directional guidance and reference for subsequent experiments on the adsorption and separation of CO_(2)/H_(2).展开更多
Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throu...Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throughput techniques.This study constructed an integrated system combining metabolic engineering and high-throughput screening of fluorescent substrates,aiming to overcome the technical bottlenecks in the identification of key enzymes for PAPS synthesis and the optimization of their catalytic efficiency.Methods:Eighteen candidate genes related to carbohydrate metabolism were screened via transcriptome sequencing of deer antler stem cells.After prokaryotic expression and preliminary screening by high-performance liquid chromatography(HPLC),a self-designed fluorescent substrate(FAM-Glc-β1-3-PNPG)combined with a 96-well plate platform was used to achieve high-throughput optimization and screening of catalytic efficiency.Enzyme function was verified through kinetic analysis and in vitro antioxidant experiments.Results:The screened high-efficiency glycosyltransferase UGT-Wnt3a showed a 2.1-fold increase in catalytic efficiency compared with the wild type,with a maximum reaction rate(Vmax)of12.3μmol·min^(-1)·mg^(-1)and a Michaelis constant(Km)of 0.87 mM.The catalytic product of UGT-Wnt3a increased the superoxide dismutase(SOD)activity by 42%(P<0.01)and decreased the malondialdehyde(MDA)content by 28%(P<0.05)in the oxidative damage system.The detection efficiency of the new platform was 9 times higher than that of HPLC,and the limit of detection was reduced by 50 times.Conclusion:The screening system established in this study provides an innovative technical solution for the directed synthesis of PAPS and the development of anti-aging components,promoting the transformation of active components in traditional Chinese medicine from natural extraction to bioengineering-based production.展开更多
For the advancement of fast-charging sodium-ion batteries(SIBs),the synthesis of cutting-edge cathode materials with superior structural stability and enhanced Na+diffusion kinetics is imperative.Multiphase layered tr...For the advancement of fast-charging sodium-ion batteries(SIBs),the synthesis of cutting-edge cathode materials with superior structural stability and enhanced Na+diffusion kinetics is imperative.Multiphase layered transition metal oxides(LTMOs),which leverage the synergistic properties of two distinct monophasic LTMOs,have garnered significant attention;however,their efficacy under fast-charging conditions remains underexplored.In this study,we developed a high-throughput computational screening framework to identify optimal dopants that maximize the electrochemical performance of LTMOs.Specifically,we evaluated the efficacy of 32 dopants based on P2/O3-type Mn/Fe-based Na_(x)Mn_(0.5)Fe_(0.5)O_(2)(NMFO)cathode material.Multiphase LTMOs satisfying criteria for thermodynamic and structural stability,minimized phase transitions,and enhanced Na^(+)diffusion were systematically screened for their suitability in fast-charging applications.The analysis identified two dopants,Ti and Zr,which met all predefined screening criteria.Furthermore,we ranked and scored dopants based on their alignment with these criteria,establishing a comprehensive dopant performance database.These findings provide a robust foundation for experimental exploration and offer detailed guidelines for tailoring dopants to optimize fast-charging SIBs.展开更多
Multiorgan-on-a-chip(MOoC)systems are advanced microfluidic devices that integrate multiple organ models into a single modular unit,each composed of cells derived from various tissues or organs.These systems enable in...Multiorgan-on-a-chip(MOoC)systems are advanced microfluidic devices that integrate multiple organ models into a single modular unit,each composed of cells derived from various tissues or organs.These systems enable interorgan communication and accurately replicate physiological conditions,providing a more physiologically relevant modeling framework for constructing disease models and predicting drug efficacy and toxicity.MOoC systems also provide significant advantages in terms of flexibility,cost-effectiveness,and reproducibility,making them valuable tools for drug development and toxicity assessment.In this review,we first provide an overview of the MOoC technology,covering cell sources,stimulations,materials and fabrication techniques,and biosensors.We then examine the application of MOoC systems in disease modeling,focusing on cancer metastasis,metabolic disorders,and cardiovascular disease.We next discuss the use of MOoC systems in drug toxicity evaluation and drug screening,emphasizing their role in providing comprehensive assessments of drug effects.Finally,we address the challenges it faces and the future perspectives of the MOoC technology.展开更多
Electrochemical nitrate(NO_(3)^(-))reduction reaction(eNO_(3)RR)presents a promising and sustainable strategy for converting environmentally hazardous NO_(3)^(-)into value-added ammonia(NH3),thereby addressing both po...Electrochemical nitrate(NO_(3)^(-))reduction reaction(eNO_(3)RR)presents a promising and sustainable strategy for converting environmentally hazardous NO_(3)^(-)into value-added ammonia(NH3),thereby addressing both pollution mitigation and nitrogen resource recovery.However,its practical implementation remains hindered by the limited availability of electrocatalysts that simultaneously offer high activity,selectivity,and stability.In this study,we systematically investigate the catalytic potential of 300 Cu-based heteronuclear trimetallic dual-atom alloys(DAAs),featuring heterometallic dual-atom active sites embedded on a catalytically suboptimal Cu(111)surface.By combining high-throughput firstprinciples calculations with a hierarchical four-step screening strategy,we efficiently identify promising DAA catalysts for eNO_(3)RR.Among them,CrRh and MnRh catalysts stand out,exhibiting ultralow limiting potentials of-0.16 and-0.25 V,respectively,along with excellent selectivity and stability.Constantpotential simulations further reveal that both catalysts maintain high activity and selectivity across varying pH conditions and applied potentials.Geometric and electronic structure analyses indicate that the incorporation of dual-atom sites induces local lattice distortion and charge polarization,effectively tuning the electronic structure of the active centers.Notably,strong d-d orbital interactions between the embedded metal dimers give rise to new molecule-like electronic states near the Fermi level.These states facilitate effective activation of NO_(3)^(-)and NO via an electron“acceptance-donation”mechanism,driven by p-d orbital interactions between adsorbates and metal dimers.This work not only strategically designs efficient heteronuclear Cu-based DAA catalysts but also provides valuable insights into the electronic synergistic effects of dual-atom sites in modulating eNO_(3)RR performance.It establishes a promising platform for demonstrating the feasibility of constructing dual-atom active centers on pure metal surfaces for eNO_(3)RR,while broadening the potential applications of DAAs in electrocatalysis and related fields.展开更多
In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment si...In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment simulation,particularly of the mechanical microenvironment,is crucial for estimating the biological effects of a tumor.However,current in vitro osteosarcoma model construction is often limited to a single mechanical signal,which fails to simulate the diversity of osteosarcoma mechanical stimuli.In this study,we utilized embedded bioprinting technology and the multiple response properties of calcium ions in soft and hard stiffness systems with osteosarcoma cell biological functions to construct an integrated gradient biomechanical signal-tailored osteosarcoma model(IGBSTOM).We achieved this by printing a fibrinogen bioink containing calcium ions and osteosarcoma tumor spheroids within an extracellular matrix composed of methacryloylated alginate,methacryloylated gelatin,thrombin,and transglutaminase,which is rich in polysaccharides and proteins and exhibits self-healing properties.Our in vitro and in vivo studies showed that the IGBSTOM enhanced tumor stemness,proliferation,and migration,and successfully reproduced the nest-like structure of tumors,providing an in vitro research platform that is more similar to the natural tumor than the existing models.This study proposes a novel IGBSTOM construction and provides a new strategy for the clinical understanding of tumor development,drug screening,and exploration of drug resistance and metastasis mechanisms.展开更多
This editorial discusses an article by Liu et al,which focuses on the development and evaluation of a modified scoring model incorporating the waist-to-hip ratio for predicting advanced colorectal neoplasia(ACN).This ...This editorial discusses an article by Liu et al,which focuses on the development and evaluation of a modified scoring model incorporating the waist-to-hip ratio for predicting advanced colorectal neoplasia(ACN).This editorial provides an overview of the study,including the background of ACN risk prediction,the study design,key findings,and the significance and limitations of the new model.The study identified independent risk factors for ACN and developed a 7-point scoring model with better predictive performance than existing models.However,challenges,such as generalizability across ethnic groups and selection bias,exist.Further research involving multi-ethnic cohorts and the integration of novel biomarkers is needed to improve the model and its clinical application.展开更多
With the continuous emergence and rapid spread of multidrug-resistant and extensively-drug-resistant Mycobacterium tuberculosis strains, it is imperative to develop novel therapies against this bacterium. The intrins...With the continuous emergence and rapid spread of multidrug-resistant and extensively-drug-resistant Mycobacterium tuberculosis strains, it is imperative to develop novel therapies against this bacterium. The intrinsic β-lactam resistance of M. tuberculosis is primarily due to the production of an Ambler class-A β-lactamase BlaC, which limits the application of β-lactam antibiotics in the treatment of tuberculosis. Therefore, the inhibitors of BlaC could be novel anti-tuberculosis drug synergistic agents to recover the sensibility of M. Tuberculosis to the β-lactam antibiotics. In the present study, BlaC of M. tuberculosis was expressed and purified to establish a screening model of the BlaC inhibitors. The screening conditions were determined, and the screening model was evaluated to fit for the high throughput screening. A total of 22 BlaC inhibitors were screened out from 26 400 compound samples with a positive rate of 0.083%. Taken together, our findings lay the foundation for the discovery of novel anti-tuberculosis drug synergistic agents in clinic.展开更多
Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment fro...Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.展开更多
The efficiency of particle screening was studied over a range of vibrational parameters including amplitude, frequency and vibrational direction. The Discrete Element Method (DEM) was used to simulate the screening pr...The efficiency of particle screening was studied over a range of vibrational parameters including amplitude, frequency and vibrational direction. The Discrete Element Method (DEM) was used to simulate the screening process. A functional relationship between efficiency and the parameters, both singly and combined, is established. The function is a complicated exponential. Optimal amplitude and frequency values are smaller for particles near the mesh and larger for other particles. The optimum vibration angle is 45° for nearly all kinds of particles. A transverse velocity, V⊥, was defined and V⊥=0.2 m/s was identified to be the most efficient operating point by both simulation and experimental observation. Comparison of these results with those reported by others is included.展开更多
Finding energetic materials with tailored properties is always a significant challenge due to low research efficiency in trial and error.Herein,a methodology combining domain knowledge,a machine learning algorithm,and...Finding energetic materials with tailored properties is always a significant challenge due to low research efficiency in trial and error.Herein,a methodology combining domain knowledge,a machine learning algorithm,and experiments is presented for accelerating the discovery of novel energetic materials.A high-throughput virtual screening(HTVS)system integrating on-demand molecular generation and machine learning models covering the prediction of molecular properties and crystal packing mode scoring is established.With the proposed HTVS system,candidate molecules with promising properties and a desirable crystal packing mode are rapidly targeted from the generated molecular space containing 25112 molecules.Furthermore,a study of the crystal structure and properties shows that the good comprehensive performances of the target molecule are in agreement with the predicted results,thus verifying the effectiveness of the proposed methodology.This work demonstrates a new research paradigm for discovering novel energetic materials and can be extended to other organic materials without manifest obstacles.展开更多
To discover new lead compounds for M1 agonists. Ten typical M1 agonists were superimposed to build a M1 agonists 3D-pharmacophore model using distance-comparisons （DISCO） method without the previous knowledge of the...To discover new lead compounds for M1 agonists. Ten typical M1 agonists were superimposed to build a M1 agonists 3D-pharmacophore model using distance-comparisons （DISCO） method without the previous knowledge of the three-dimensional structure of M1 receptor. Virtual screening strategy was used to analyze the Available Chemicals Directory-Screening Compounds （ACD-SC） to identify possible new hits. Twenty-two compounds which fit the pharmacophore model well and are not similar with known M1 agonists were purchased in order to evaluate their M1 receptor agonist activity. One of them shows M1 receptor agonist activity with EC50 of 4.90 μmol/L and maximum response. Multiple of 10.0 which shows it worthy of further study as a new lead compound for M1 agonists.展开更多
Natural antimicrobial peptides(AMPs)are promising candidates for the development of a new generation of antimicrobials to combat antibiotic-resistant pathogens.They have found extensive applications in the fields of m...Natural antimicrobial peptides(AMPs)are promising candidates for the development of a new generation of antimicrobials to combat antibiotic-resistant pathogens.They have found extensive applications in the fields of medicine,food,and agriculture.However,efficiently screening AMPs from natural sources poses several challenges,including low efficiency and high antibiotic resistance.This review focuses on the action mechanisms of AMPs,both through membrane and non-membrane routes.We thoroughly examine various highly efficient AMP screening methods,including whole-bacterial adsorption binding,cell membrane chromatography(CMC),phospholipid membrane chromatography binding,membranemediated capillary electrophoresis(CE),colorimetric assays,thin layer chromatography(TLC),fluorescence-based screening,genetic sequencing-based analysis,computational mining of AMP databases,and virtual screening methods.Additionally,we discuss potential developmental applications for enhancing the efficiency of AMP discovery.This review provides a comprehensive framework for identifying AMPs within complex natural product systems.展开更多
Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening （HTS） data for 3 762 distinct global gene knockout （KO） mouse lines with viable adult hom...Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening （HTS） data for 3 762 distinct global gene knockout （KO） mouse lines with viable adult homozygous mice generated using either gene-trap or homologous recombination technologies. Bone mass was determined from DEXA scans of male and female mice at 14 weeks of age and by microCT analyses of bones from male mice at 16 weeks of age. Wild-type （WT） cagemates/littermates were examined for each gene KO. Lethality was observed in an additional 850 KO lines. Since primary HTS are susceptible to false positive findings, additional cohorts of mice from KO lines with intriguing HTS bone data were examined. Aging, ovariectomy, histomorphometry and bone strength studies were performed and possible non-skeletal phenotypes were explored. Together, these screens identified multiple genes affecting bone mass： 23 previously reported genes （Calcr, Cebpb, Crtap, Dcstamp, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.展开更多
Ion channels are attractive targets for drug discovery as an increasing number of new ion channel targets have been uncovered in diseases, such as pain, cardiovascular disease, and neurological disorders. Despite thei...Ion channels are attractive targets for drug discovery as an increasing number of new ion channel targets have been uncovered in diseases, such as pain, cardiovascular disease, and neurological disorders. Despite their relevance in diseases and the variety of physiological functions they are involved in, ion channels still remain underexploited as drug targets. This, to a large extent, is attributed to the absence of screening technologies that ensure both the quality and the throughput of data. However, an increasing number of assays and technologies have evolved rapidly in the past decades. In this review, we summarized the currently available high-throughput screening technologies in ion channel drug discovery.展开更多
Objective To develop a high-throughput screening assay for Farnesoid X receptor （FXR） agonists based on mammalian one-hybrid system （a chimera receptor gene system） for the purpose of identifying new lead compound...Objective To develop a high-throughput screening assay for Farnesoid X receptor （FXR） agonists based on mammalian one-hybrid system （a chimera receptor gene system） for the purpose of identifying new lead compounds for dyslipidaemia drug from the chemical library. Methods cDNA encoding the human FXR ligand binding domain （LBD） was amplified by RT-PCR from a human liver total mRNA and fused to the DNA binding domain （DBD） of yeast GAL4 of pBIND to construct a GAL4-FXR （LBD） chimera expression plasmid. Five copies of the GAL4 DNA binding site were synthesized and inserted into upstream of the SV40 promoter of pGL3-promoter vector to construct a reporter plasmid pG5-SV40 Luc. The assay was developed by transient co-transfection with pG5-SV40 Luc reporter plasmid and pBIND-FXR-LBD （189-472） chimera expression plasmid. Results After optimization, CDCA, a FXR natural agonist, could induce expression of the luciferase gene in a dose-dependent manner, and had a signal/noise ratio of 10 and Z＇ factor value of 0.65, Conclusion A stable and sensitive cell-based high-throughput screening model can be used in high-throughput screening for FXR agonists from the synthetic and natural compound library.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.39930190)Meg a-projects of Science Rescarch for the 10th Five-Year Plan(Grant No.2004AA2Z3784).
文摘To develop a new high-throughput screening model for human high-density lipoprotein(HDL)receptor(CD36 and LIMPⅡanalogous-1,CLA-1)agonists using CLA-1-expressing insect cells.Methods With the total RNA of human hepatoma cells BEL-7402 as template,the complementary DNA(cDNA)of CLA-1 was amplified by reverse transcription-polymerase chain reaction(RT-PCR).Bac-to-Bac baculovirus expression system was used to express CLA-1 in insect cells.CLA-1 cDNA was cloned downstream of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus(AcNPV)into donor vector pFastBacl and recombinant pFastBacl-CLA-1 was transformed into E.coli DH10Bac to transpose CLA-1 cDNA to bacrnid DNA.Recombinant bacrnid-CLA-1 was transfected into Spodopterafrugiperda Sf9 insect cells to produce recombinant baculovirus particles.Recombinant CLA-1 was expressed on the membrane of Sf9 cells infected with the recombinant baculoviruses.A series of parameters of DiI-lipoprotein binding assays of CLA-1-expressing Sf9 cells in 96-well plates were optimized.Results Western blot analysis and DiI-lipoprotein binding assays confirmed that CLA-1 expressed in insect cells had similar immunoreactivity and ligand binding activity as its native counterpart.A reliable and sensitive in vitro cell-based assay was established to assess the activity of CLA-1 and used to screen agonists from different sample libraries.Conclusion Human HDL receptor CLA-1 was successfully expressed in Sf9 insect cells and a novel high-throughput screening model for CLA-1 agonists was developed.Utilization of this model allows us to identify potent and selective CLA-1 agonists which might possibly be used as therapeutics for atherosclerosis.
基金financial support from the National Key Research and Development Program of China(2021YFB 3501501)the National Natural Science Foundation of China(No.22225803,22038001,22108007 and 22278011)+1 种基金Beijing Natural Science Foundation(No.Z230023)Beijing Science and Technology Commission(No.Z211100004321001).
文摘The high porosity and tunable chemical functionality of metal-organic frameworks(MOFs)make it a promising catalyst design platform.High-throughput screening of catalytic performance is feasible since the large MOF structure database is available.In this study,we report a machine learning model for high-throughput screening of MOF catalysts for the CO_(2) cycloaddition reaction.The descriptors for model training were judiciously chosen according to the reaction mechanism,which leads to high accuracy up to 97%for the 75%quantile of the training set as the classification criterion.The feature contribution was further evaluated with SHAP and PDP analysis to provide a certain physical understanding.12,415 hypothetical MOF structures and 100 reported MOFs were evaluated under 100℃ and 1 bar within one day using the model,and 239 potentially efficient catalysts were discovered.Among them,MOF-76(Y)achieved the top performance experimentally among reported MOFs,in good agreement with the prediction.
基金supported by the National Key Research and Development Program of China(No.2021YFB3702102)support from the“Initiation Program for New Teachers”(No.AF0500207)+1 种基金Shanghai Jiao Tong Universitysupport from the Changsha Science and Technology Plan International and Regional Cooperation Project(No.kh2304002)。
文摘Lithium-ion batteries(LiBs)with high energy density have gained significant popularity in smart grids and portable electronics.LiMn_(1-x)Fe_(x)PO_(4)(LMFP)is considered a leading candidate for the cathode,with the potential to combine the low cost of Li Fe PO_(4)(LFP)with the high theoretical energy density of LiMnPO_(4)(LMP).However,quantitative investigation of the intricate coupling between the Fe/Mn ratio and the resulting energy density is challenging due to the parametric complexity.It is crucial to develop a universal approach for the rapid construction of multi-parameter mapping.In this work,we propose an active learning-guided high-throughput workflow for quantitatively predicting the Fe/Mn ratio and the energy density mapping of LMFP.An optimal composition(LiMn_(0.66)Fe_(0.34)PO_(4))was effectively screened from 81 cathode materials via only 5 samples.Model-guided electrochemical analysis revealed a nonlinear relationship between the Fe/Mn ratio and electrochemical properties,including ion mobility and impedance,elucidating the quantitative chemical composition-energy density map of LMFP.The results demonstrated the efficacy of the method in high-throughput screening of LiBs cathode materials.
基金supported by the Shandong Provincial Key Research and Development Program(Major Key Technology Project)(2021CXGC010514)the National Natural Science Foundation of China(22008173).
文摘Additives are widely employed to regulate the morphology,size,and agglomeration degree of crystalline materials during crystallization to enhance their functional,physical,and powder properties.However,the existing methods for screening and validating target additives require a large quantity of materials and involve tedious molecular simulation/crystallization experiments,making them time-consuming,resource-intensive,and reliant on the operator’s experience level.To overcome these challenges,we proposed a computer vision-assisted high-throughput additive screening system(CV-HTPASS)which comprises a high-throughput additive screening device,in situ imaging equipment,and an artificial intelligence(AI)-assisted image-analysis algorithm.Using the CV-HTPASS,we performed high-throughput screening experiments on additives to regulate the succinic acid crystal properties,generating thousands of crystal images with diverse crystal morphologies.To extract valuable crystal information from the massive data and improve the analysis accuracy and efficiency,the AI-based image-analysis algorithm was implemented innovatively for the segmentation,classification,and data mining of crystals with four morphologies to further screen the target additive.Subsequently,scale-up crystallization experiments conducted under optimized conditions demonstrated that succinic acid products exhibited a preferred cubic morphology,reduced agglomeration degree,narrowed crystal size distribution,and improved powder properties.The proposed CV-HTPASS offers a highly efficient approach for scale-up experiments.Further,it provides a platform for the screening of additives and the optimization of the powder properties of crystal products in industrial-scale crystallization processes.
基金Project supported by the National Natural Science Foundation of China(Grant Nos.11304079,11404094,and 11504088)Science and Technology Research Project of Henan Science and Technology Department(Grant No.182102410076)。
文摘The capture of CO_(2)from CO_(2)/H_(2)gas mixtures in syngas is a crucial issue for hydrogen production from steam methane reforming in industry,as the presence of CO_(2)directly affects the purity of H_(2).A combination of a high-throughput screening method and grand canonical Monte Carlo simulation was utilized to evaluate and screen 1725 metal–organic frameworks(MOFs)in detail as a means of determining their adsorption performance for CO_(2)/H_(2)gas mixtures.The adsorption and separation performance of double-linker MOFs was comprehensively evaluated using eight evaluation indicators,namely,the largest cavity diameter,accessible surface area,pore occupied accessible volume,porosity,adsorption selectivity,working capacity,adsorbent performance score and percent regeneration.Six optimal performance frameworks were screened to further study their single-component adsorption and binary competitive adsorption of CO_(2)/H_(2)respectively.The CO_(2)adsorption selectivity at different CO_(2)/H_(2)feed ratios was also evaluated,which indicated their excellent adsorption and separation performance.The microscopic adsorption mechanisms for CO_(2)and H_(2)at the molecular level were investigated by analyzing the radial distribution function and density distribution.This study may provide directional guidance and reference for subsequent experiments on the adsorption and separation of CO_(2)/H_(2).
文摘Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throughput techniques.This study constructed an integrated system combining metabolic engineering and high-throughput screening of fluorescent substrates,aiming to overcome the technical bottlenecks in the identification of key enzymes for PAPS synthesis and the optimization of their catalytic efficiency.Methods:Eighteen candidate genes related to carbohydrate metabolism were screened via transcriptome sequencing of deer antler stem cells.After prokaryotic expression and preliminary screening by high-performance liquid chromatography(HPLC),a self-designed fluorescent substrate(FAM-Glc-β1-3-PNPG)combined with a 96-well plate platform was used to achieve high-throughput optimization and screening of catalytic efficiency.Enzyme function was verified through kinetic analysis and in vitro antioxidant experiments.Results:The screened high-efficiency glycosyltransferase UGT-Wnt3a showed a 2.1-fold increase in catalytic efficiency compared with the wild type,with a maximum reaction rate(Vmax)of12.3μmol·min^(-1)·mg^(-1)and a Michaelis constant(Km)of 0.87 mM.The catalytic product of UGT-Wnt3a increased the superoxide dismutase(SOD)activity by 42%(P<0.01)and decreased the malondialdehyde(MDA)content by 28%(P<0.05)in the oxidative damage system.The detection efficiency of the new platform was 9 times higher than that of HPLC,and the limit of detection was reduced by 50 times.Conclusion:The screening system established in this study provides an innovative technical solution for the directed synthesis of PAPS and the development of anti-aging components,promoting the transformation of active components in traditional Chinese medicine from natural extraction to bioengineering-based production.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korean government(MSIT)(No.2022R1F1A1074339)。
文摘For the advancement of fast-charging sodium-ion batteries(SIBs),the synthesis of cutting-edge cathode materials with superior structural stability and enhanced Na+diffusion kinetics is imperative.Multiphase layered transition metal oxides(LTMOs),which leverage the synergistic properties of two distinct monophasic LTMOs,have garnered significant attention;however,their efficacy under fast-charging conditions remains underexplored.In this study,we developed a high-throughput computational screening framework to identify optimal dopants that maximize the electrochemical performance of LTMOs.Specifically,we evaluated the efficacy of 32 dopants based on P2/O3-type Mn/Fe-based Na_(x)Mn_(0.5)Fe_(0.5)O_(2)(NMFO)cathode material.Multiphase LTMOs satisfying criteria for thermodynamic and structural stability,minimized phase transitions,and enhanced Na^(+)diffusion were systematically screened for their suitability in fast-charging applications.The analysis identified two dopants,Ti and Zr,which met all predefined screening criteria.Furthermore,we ranked and scored dopants based on their alignment with these criteria,establishing a comprehensive dopant performance database.These findings provide a robust foundation for experimental exploration and offer detailed guidelines for tailoring dopants to optimize fast-charging SIBs.
基金supported by the National Natural Science Foundation of China(No.32371475)the Natural Science Foundation of Jiangsu Province,Major Project(No.BK20222008).
文摘Multiorgan-on-a-chip(MOoC)systems are advanced microfluidic devices that integrate multiple organ models into a single modular unit,each composed of cells derived from various tissues or organs.These systems enable interorgan communication and accurately replicate physiological conditions,providing a more physiologically relevant modeling framework for constructing disease models and predicting drug efficacy and toxicity.MOoC systems also provide significant advantages in terms of flexibility,cost-effectiveness,and reproducibility,making them valuable tools for drug development and toxicity assessment.In this review,we first provide an overview of the MOoC technology,covering cell sources,stimulations,materials and fabrication techniques,and biosensors.We then examine the application of MOoC systems in disease modeling,focusing on cancer metastasis,metabolic disorders,and cardiovascular disease.We next discuss the use of MOoC systems in drug toxicity evaluation and drug screening,emphasizing their role in providing comprehensive assessments of drug effects.Finally,we address the challenges it faces and the future perspectives of the MOoC technology.
基金supported by the Water Conservancy Science and Technology Project of Jiangsu Province,China(No.2024005,2025012)。
文摘Electrochemical nitrate(NO_(3)^(-))reduction reaction(eNO_(3)RR)presents a promising and sustainable strategy for converting environmentally hazardous NO_(3)^(-)into value-added ammonia(NH3),thereby addressing both pollution mitigation and nitrogen resource recovery.However,its practical implementation remains hindered by the limited availability of electrocatalysts that simultaneously offer high activity,selectivity,and stability.In this study,we systematically investigate the catalytic potential of 300 Cu-based heteronuclear trimetallic dual-atom alloys(DAAs),featuring heterometallic dual-atom active sites embedded on a catalytically suboptimal Cu(111)surface.By combining high-throughput firstprinciples calculations with a hierarchical four-step screening strategy,we efficiently identify promising DAA catalysts for eNO_(3)RR.Among them,CrRh and MnRh catalysts stand out,exhibiting ultralow limiting potentials of-0.16 and-0.25 V,respectively,along with excellent selectivity and stability.Constantpotential simulations further reveal that both catalysts maintain high activity and selectivity across varying pH conditions and applied potentials.Geometric and electronic structure analyses indicate that the incorporation of dual-atom sites induces local lattice distortion and charge polarization,effectively tuning the electronic structure of the active centers.Notably,strong d-d orbital interactions between the embedded metal dimers give rise to new molecule-like electronic states near the Fermi level.These states facilitate effective activation of NO_(3)^(-)and NO via an electron“acceptance-donation”mechanism,driven by p-d orbital interactions between adsorbates and metal dimers.This work not only strategically designs efficient heteronuclear Cu-based DAA catalysts but also provides valuable insights into the electronic synergistic effects of dual-atom sites in modulating eNO_(3)RR performance.It establishes a promising platform for demonstrating the feasibility of constructing dual-atom active centers on pure metal surfaces for eNO_(3)RR,while broadening the potential applications of DAAs in electrocatalysis and related fields.
基金appreciate financial support from the National Key R&D Program of China(No.2022YFA1104600)2022 Lingang Laboratory“Seeking Outstanding Youth Program”Open Project(No.LGQS-202206-04)+3 种基金Shanghai Ninth People’s Hospital–Shanghai Jiao Tong University School of Medicine–Shanghai University of Science and Technology Cross-funded Collaborative Program(No.JYJC202233)the National Natural Science Foundation of China(No.82372377)Biomaterials and Regenerative Medicine Institute Cooperative Research Project by Shanghai Jiao Tong University School of Medicine(No.2022LHBO8),Shanghai Key Laboratory of Orthopaedic Implants,Department of Orthopaedics by Shanghai Ninth People’s Hospital–Shanghai Jiao Tong University School of Medicine(No.KFKT202206),the Key R&D Program of Jiangsu Province Social Development Project(No.BE2022708)the Project of Shanghai Science and Technology Commission(No.22015820100).
文摘In the current settings of osteosarcoma research and drug screening,in vitro three-dimensional(3D)models,which overcome the limitations of traditional models,are favored.In in vitro 3D models,tumor microenvironment simulation,particularly of the mechanical microenvironment,is crucial for estimating the biological effects of a tumor.However,current in vitro osteosarcoma model construction is often limited to a single mechanical signal,which fails to simulate the diversity of osteosarcoma mechanical stimuli.In this study,we utilized embedded bioprinting technology and the multiple response properties of calcium ions in soft and hard stiffness systems with osteosarcoma cell biological functions to construct an integrated gradient biomechanical signal-tailored osteosarcoma model(IGBSTOM).We achieved this by printing a fibrinogen bioink containing calcium ions and osteosarcoma tumor spheroids within an extracellular matrix composed of methacryloylated alginate,methacryloylated gelatin,thrombin,and transglutaminase,which is rich in polysaccharides and proteins and exhibits self-healing properties.Our in vitro and in vivo studies showed that the IGBSTOM enhanced tumor stemness,proliferation,and migration,and successfully reproduced the nest-like structure of tumors,providing an in vitro research platform that is more similar to the natural tumor than the existing models.This study proposes a novel IGBSTOM construction and provides a new strategy for the clinical understanding of tumor development,drug screening,and exploration of drug resistance and metastasis mechanisms.
文摘This editorial discusses an article by Liu et al,which focuses on the development and evaluation of a modified scoring model incorporating the waist-to-hip ratio for predicting advanced colorectal neoplasia(ACN).This editorial provides an overview of the study,including the background of ACN risk prediction,the study design,key findings,and the significance and limitations of the new model.The study identified independent risk factors for ACN and developed a 7-point scoring model with better predictive performance than existing models.However,challenges,such as generalizability across ethnic groups and selection bias,exist.Further research involving multi-ethnic cohorts and the integration of novel biomarkers is needed to improve the model and its clinical application.
基金Fundamental Research Funds for Central Public Welfare Research Institutes(Grant No.2015PT350001)National Major Scientific and Technological Special Project for “Significant New Drugs Development”(Grant No.2015ZX09102007-009)
文摘With the continuous emergence and rapid spread of multidrug-resistant and extensively-drug-resistant Mycobacterium tuberculosis strains, it is imperative to develop novel therapies against this bacterium. The intrinsic β-lactam resistance of M. tuberculosis is primarily due to the production of an Ambler class-A β-lactamase BlaC, which limits the application of β-lactam antibiotics in the treatment of tuberculosis. Therefore, the inhibitors of BlaC could be novel anti-tuberculosis drug synergistic agents to recover the sensibility of M. Tuberculosis to the β-lactam antibiotics. In the present study, BlaC of M. tuberculosis was expressed and purified to establish a screening model of the BlaC inhibitors. The screening conditions were determined, and the screening model was evaluated to fit for the high throughput screening. A total of 22 BlaC inhibitors were screened out from 26 400 compound samples with a positive rate of 0.083%. Taken together, our findings lay the foundation for the discovery of novel anti-tuberculosis drug synergistic agents in clinic.
文摘Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.
基金the Special Topic of Key Science and Technology of Fujian Province Fund (No.2006HZ0002-2)
文摘The efficiency of particle screening was studied over a range of vibrational parameters including amplitude, frequency and vibrational direction. The Discrete Element Method (DEM) was used to simulate the screening process. A functional relationship between efficiency and the parameters, both singly and combined, is established. The function is a complicated exponential. Optimal amplitude and frequency values are smaller for particles near the mesh and larger for other particles. The optimum vibration angle is 45° for nearly all kinds of particles. A transverse velocity, V⊥, was defined and V⊥=0.2 m/s was identified to be the most efficient operating point by both simulation and experimental observation. Comparison of these results with those reported by others is included.
基金the Science Challenge Project(TZ2018004)the National Natural Science Foundation of China(21875228 and 21702195)for financial support。
文摘Finding energetic materials with tailored properties is always a significant challenge due to low research efficiency in trial and error.Herein,a methodology combining domain knowledge,a machine learning algorithm,and experiments is presented for accelerating the discovery of novel energetic materials.A high-throughput virtual screening(HTVS)system integrating on-demand molecular generation and machine learning models covering the prediction of molecular properties and crystal packing mode scoring is established.With the proposed HTVS system,candidate molecules with promising properties and a desirable crystal packing mode are rapidly targeted from the generated molecular space containing 25112 molecules.Furthermore,a study of the crystal structure and properties shows that the good comprehensive performances of the target molecule are in agreement with the predicted results,thus verifying the effectiveness of the proposed methodology.This work demonstrates a new research paradigm for discovering novel energetic materials and can be extended to other organic materials without manifest obstacles.
基金National Natural Science Foundation of China (Grant No. 30271538)985 program,Ministry of Education of China
文摘To discover new lead compounds for M1 agonists. Ten typical M1 agonists were superimposed to build a M1 agonists 3D-pharmacophore model using distance-comparisons （DISCO） method without the previous knowledge of the three-dimensional structure of M1 receptor. Virtual screening strategy was used to analyze the Available Chemicals Directory-Screening Compounds （ACD-SC） to identify possible new hits. Twenty-two compounds which fit the pharmacophore model well and are not similar with known M1 agonists were purchased in order to evaluate their M1 receptor agonist activity. One of them shows M1 receptor agonist activity with EC50 of 4.90 μmol/L and maximum response. Multiple of 10.0 which shows it worthy of further study as a new lead compound for M1 agonists.
基金supported by the National Natural Science Foundation of China(Grant Nos.:82373835,82304437,and 82173781)Regional Joint Fund Project of Guangdong Basic and Applied Basic Research Fund,China(Grant Nos.:2023A1515110417 and 2023A1515140131)+2 种基金Regional Joint Fund-Key Project of Guangdong Basic and Applied Basic Research Fund,China(Grant No.:2020B1515120033)the Key Field Projects of General Universities in Guangdong Province,China(Grant Nos.:2020ZDZX2057 and 2022ZDZX2056)Medical Scientific Research Foundation of Guangdong Province of China(Grant No.:A2022061).
文摘Natural antimicrobial peptides(AMPs)are promising candidates for the development of a new generation of antimicrobials to combat antibiotic-resistant pathogens.They have found extensive applications in the fields of medicine,food,and agriculture.However,efficiently screening AMPs from natural sources poses several challenges,including low efficiency and high antibiotic resistance.This review focuses on the action mechanisms of AMPs,both through membrane and non-membrane routes.We thoroughly examine various highly efficient AMP screening methods,including whole-bacterial adsorption binding,cell membrane chromatography(CMC),phospholipid membrane chromatography binding,membranemediated capillary electrophoresis(CE),colorimetric assays,thin layer chromatography(TLC),fluorescence-based screening,genetic sequencing-based analysis,computational mining of AMP databases,and virtual screening methods.Additionally,we discuss potential developmental applications for enhancing the efficiency of AMP discovery.This review provides a comprehensive framework for identifying AMPs within complex natural product systems.
文摘Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening （HTS） data for 3 762 distinct global gene knockout （KO） mouse lines with viable adult homozygous mice generated using either gene-trap or homologous recombination technologies. Bone mass was determined from DEXA scans of male and female mice at 14 weeks of age and by microCT analyses of bones from male mice at 16 weeks of age. Wild-type （WT） cagemates/littermates were examined for each gene KO. Lethality was observed in an additional 850 KO lines. Since primary HTS are susceptible to false positive findings, additional cohorts of mice from KO lines with intriguing HTS bone data were examined. Aging, ovariectomy, histomorphometry and bone strength studies were performed and possible non-skeletal phenotypes were explored. Together, these screens identified multiple genes affecting bone mass： 23 previously reported genes （Calcr, Cebpb, Crtap, Dcstamp, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.
基金supported by the State Key Laboratory of Natural and Biomimetic Drugs, Peking University。
文摘Ion channels are attractive targets for drug discovery as an increasing number of new ion channel targets have been uncovered in diseases, such as pain, cardiovascular disease, and neurological disorders. Despite their relevance in diseases and the variety of physiological functions they are involved in, ion channels still remain underexploited as drug targets. This, to a large extent, is attributed to the absence of screening technologies that ensure both the quality and the throughput of data. However, an increasing number of assays and technologies have evolved rapidly in the past decades. In this review, we summarized the currently available high-throughput screening technologies in ion channel drug discovery.
基金supported by the Ministry of Science and Technology, PRC in Mega-projects of Science Research During the 10th Five-Year Plan Period (No. 2004AA2Z38784)National Natural Science Foundation of China (No. 30472026).
文摘Objective To develop a high-throughput screening assay for Farnesoid X receptor （FXR） agonists based on mammalian one-hybrid system （a chimera receptor gene system） for the purpose of identifying new lead compounds for dyslipidaemia drug from the chemical library. Methods cDNA encoding the human FXR ligand binding domain （LBD） was amplified by RT-PCR from a human liver total mRNA and fused to the DNA binding domain （DBD） of yeast GAL4 of pBIND to construct a GAL4-FXR （LBD） chimera expression plasmid. Five copies of the GAL4 DNA binding site were synthesized and inserted into upstream of the SV40 promoter of pGL3-promoter vector to construct a reporter plasmid pG5-SV40 Luc. The assay was developed by transient co-transfection with pG5-SV40 Luc reporter plasmid and pBIND-FXR-LBD （189-472） chimera expression plasmid. Results After optimization, CDCA, a FXR natural agonist, could induce expression of the luciferase gene in a dose-dependent manner, and had a signal/noise ratio of 10 and Z＇ factor value of 0.65, Conclusion A stable and sensitive cell-based high-throughput screening model can be used in high-throughput screening for FXR agonists from the synthetic and natural compound library.
