AIM:The study aims to investigate the effects of Huayu-Qutan formula(HYQT)-medicated serum on oleic acid-induced lipid damage and endoplasmic reticulum stress(ERS)in HepG2 cells,and to explore the mecha-nism underlying the prevention and treatment of lipid metabolism disorders by HYQT.METHODS:The HepG2 cells were treated with various concentrations of oleic acid,and CCK8 assay,oil red O staining and ELISA were used to identify the optimal treatment concentration and time of oleic acid and HYQT-medicated serum.Moreover,the cells were divided into control,model,thapsigargin(Tg),and Tg+HYQT groups.Cellular lipid deposition was measured using oil red O staining,while triglyceride(TG)and free fatty acid(FFA)levels were assessed by ELISA.Transmission electron micros-copy was used to examine endoplasmic reticulum structures,and RT-qPCR and Wes fully automated protein quantification analysis system were used to measure the mRNA and protein expression of glucose-regulated protein 78(GRP78),sterol regulatory element binding protein 1c(SREBP1c),acetyl coenzyme A carboxylase(ACC1),fatty acid synthase(FAS)and stearoyl coenzyme A desaturase 1(SCD1)in different groups.RESULTS:Significant lipid deposition was induced in HepG2 cells after treatment with 1 000 μmol/L oleic acid,while treatment with serum containing 10%HYQT for 48 h was found to be optimal.Compared with control group,the cells in model group showed significant deposition of oil red O-stained lipid droplets in the cytoplasm,associated with endoplasmic reticulum expansion,ERS,and nuclear condensa-tion.The TG and FFA levels,and the mRNA and protein expression levels of GRP78,SREBP1c,ACC1,FAS and SCD1 were increased significantly(P<0.01).Compared with model group,the cells treated with HYQT-mediated serum showed marked decrease in the number of lipid droplets in the cytoplasm,with restored endoplasmic reticulum morphology.The TG and FFA levels were significantly reduced(P<0.01),and the mRNA and protein levels of factors related to ERS and de novo fatty acid synthesis were markedly decreased(P<0.01).Treatment with Tg,an ERS agonist,led to greater accu-mulation of cytoplasmic lipid droplets and increased endoplasmic reticulum expansion.Marked variations in the morpholo-gy and size of the endoplasmic reticulum were observed,with fusion and clustering of vacuoles.The TG and FFA levels,and the expression of ERS-and fatty acid synthesis-related factors were increased(P<0.01).Compared with Tg group,the cells treated with Tg+HYQT showed reduced number of cytoplasmic lipid droplets,attenuated endoplasmic reticulum dilation,and decreased number and volume of vacuoles,while the TG and FFA levels and the expression of ERS-and fatty acid synthesis-associated factors were significantly decreased(P<0.01).CONCLUSION:Serum containing HYQT alle-viates oleic acid-induced lipid damage in HepG2 cells by inhibiting ERS-induced de novo synthesis of fatty acids.

AIM:The study aims to investigate the effects of Huayu-Qutan formula(HYQT)-medicated serum on oleic acid-induced lipid damage and endoplasmic reticulum stress(ERS)in HepG2 cells,and to explore the mecha-nism underlying the prevention and treatment of lipid metabolism disorders by HYQT.METHODS:The HepG2 cells were treated with various concentrations of oleic acid,and CCK8 assay,oil red O staining and ELISA were used to identify the optimal treatment concentration and time of oleic acid and HYQT-medicated serum.Moreover,the cells were divided into control,model,thapsigargin(Tg),and Tg+HYQT groups.Cellular lipid deposition was measured using oil red O staining,while triglyceride(TG)and free fatty acid(FFA)levels were assessed by ELISA.Transmission electron micros-copy was used to examine endoplasmic reticulum structures,and RT-qPCR and Wes fully automated protein quantification analysis system were used to measure the mRNA and protein expression of glucose-regulated protein 78(GRP78),sterol regulatory element binding protein 1c(SREBP1c),acetyl coenzyme A carboxylase(ACC1),fatty acid synthase(FAS)and stearoyl coenzyme A desaturase 1(SCD1)in different groups.RESULTS:Significant lipid deposition was induced in HepG2 cells after treatment with 1 000 μmol/L oleic acid,while treatment with serum containing 10%HYQT for 48 h was found to be optimal.Compared with control group,the cells in model group showed significant deposition of oil red O-stained lipid droplets in the cytoplasm,associated with endoplasmic reticulum expansion,ERS,and nuclear condensa-tion.The TG and FFA levels,and the mRNA and protein expression levels of GRP78,SREBP1c,ACC1,FAS and SCD1 were increased significantly(P<0.01).Compared with model group,the cells treated with HYQT-mediated serum showed marked decrease in the number of lipid droplets in the cytoplasm,with restored endoplasmic reticulum morphology.The TG and FFA levels were significantly reduced(P<0.01),and the mRNA and protein levels of factors related to ERS and de novo fatty acid synthesis were markedly decreased(P<0.01).Treatment with Tg,an ERS agonist,led to greater accu-mulation of cytoplasmic lipid droplets and increased endoplasmic reticulum expansion.Marked variations in the morpholo-gy and size of the endoplasmic reticulum were observed,with fusion and clustering of vacuoles.The TG and FFA levels,and the expression of ERS-and fatty acid synthesis-related factors were increased(P<0.01).Compared with Tg group,the cells treated with Tg+HYQT showed reduced number of cytoplasmic lipid droplets,attenuated endoplasmic reticulum dilation,and decreased number and volume of vacuoles,while the TG and FFA levels and the expression of ERS-and fatty acid synthesis-associated factors were significantly decreased(P<0.01).CONCLUSION:Serum containing HYQT alle-viates oleic acid-induced lipid damage in HepG2 cells by inhibiting ERS-induced de novo synthesis of fatty acids.

Widespread utilization of glyphosate has led to environmental residues,posing potential threats to ecological systems and human health.Traditional methods for detection of glyphosate are limited by specialized equipment and operational techniques,resulting in inefficient responses.Therefore,it is urgent to develop a convenient,sensitive and accurate detection method for detection of glyphosate.Herein,a new naphthalenesulfonamide-based"Turn-on"fluorescent probe was synthesized using 2-chloroaniline and dansyl chloride as raw materials through a one-step process,which showed a good linear relationship between the glyphosate concentration in concentration range of 0.003-70 μmol/L and the fluorescence intensity(R2=0.995),with a detection limit of 2.73 nmol/L(S/N=3).Analytical techniques such as nuclear magnetic resonance(NMR)spectroscopy and high-resolution mass spectrometry(HRMS)were used to investigate the interaction mechanism between the fluorescent probe and glyphosate.The results indicated that a nucleophilic substitution reaction occurred between the probe and the secondary amine(—NH—)of glyphosate,inducing a photoinduced electron transfer(PET)effect which enhanced the fluorescence intensity by 11.2 times.The probe showed good anti-interference ability towards coexisting metal ions,anions and pesticides in water.When applied to determination of glyphosate in the samples such as tap water,river water(Xiangxi River Reservoir),soil,soybeans,and corn,the spiking recoveries ranged from 94.7％to 109.9％,demonstrating the high accuracy and broad applicability of this detection method.A portable test strip based on this fluorescent probe was developed for rapid semi-quantitative analysis of glyphosate.The developed method was rapid,sensitive,and portable,providing theoretical and technical support for on-site measurement of environmental contaminants.

ObjectiveThe detection of RNA single nucleotide polymorphism (SNP) is of great importance due to their association with protein expression related to various diseases and drug responses. At present, splintR ligase-assisted methods are important approaches for RNA direct detection, but its specificity will be limited when the fidelity of ligases is not ideal. The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection. MethodsIn this study, a dual-competitive-padlock-probe (DCPLP) assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation. To verify the method, we employed dual competitive padlock probe-mediated rolling circle amplification (DCPLP-RCA) to genotype the CYP2C9 gene. ResultsThe specificity was well improved through the competition and strand displacement of dual padlock probe, with an 83.26% reduction in nonspecific signal. By detecting synthetic RNA samples, the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L. Furthermore, clinical samples were applied to the method to evaluate its performance, and the genotyping results were consistent with those obtained using the qPCR method. ConclusionThis study has successfully established a highly specific direct RNA SNP detection method, and provided a novel avenue for accurate identification of various types of RNAs.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.

Objective:To construct a set of rationality evaluation index system of medical service price items,so as to provide references and tools for improving the mechanism of finding abnormal medical service price items and the mechanism of monitoring and evaluating medical service price items.Methods:Literature Review,focus group discussion,Delphi method and Analytic Hierarchy Process were used to determine the evaluation indexes and their weights.Results:The rationality evaluation index system of medical service price items included 5 primary indexes,10 secondary indexes and 22 third-level indexes.According to the weight,primary indexes were cost compensation(0.253),clinical use(0.227),price comparison relationship(0.225),reform guidance(0.164)and technical evaluation(0.131).The positive coefficient of experts,1evel of authority and coordination of opinions were confirmed.The items whose specific single index score was zero or whose total score was below the threshold were considered as abnormal items.Conclusion:It constructed a set of rationality evaluation index system of medical service price items,which has high scientificity and reliability.Due to the prerequisites for obtaining the basic data required for the evaluation,it is recommended that a pilot project be carried out in the pilot areas of deepening the reform of medical service prices for the purpose of further verifying and optimizing the index system.