The flavonoids from Pentorum chinense Pursh.mediates ferroptosis to alleviate sorafenib-induced liver injury in BRL-3A cells

Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(PCP)has hepatoprotective properties.There are currently few reports on PCP’s protective impact and mechanism against sorafenib-induced liver injury.Methods:To create a liver injury model,sorafenib was administered to BRL-3A cells.Cell viability assays,immunofluorescence tests,Western blotting,real-time quantitative PCR,and high-content imaging systems were utilized to examine PCP’s effect and mechanism.Results:In this study,PCP treatment mitigated the liver damage caused by sorafenib by enhancing cell survival,lowering lipid reactive oxygen species and malondialdehyde levels,and elevating glutathione levels.In addition,PCP can enhance the protein expression of cystine/glutamate transporter xCT and glutathione peroxidase 4,reduce iron content and alleviate mitochondrial toxicity.Further mechanism studies revealed that PCP inhibited ferroptosis by promoting the production of nuclear factor E2-related factor 2 nuclear translocation and subsequently affecting target genes(HO-1 and NQO1).Conclusion:Together,PCP regulates the nuclear factor E2-related factor 2 pathway,which helps to lessen ferroptosis brought on by sorafenib.

Automatic analytical approach for the determination of 12 illicit drugs and nicotine metabolites in wastewater using on-line SPE-UHPLC-MS/MS

In this study,we developed a novel on-line solid phase extraction(SPE)-ultra-high-performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS)-based analytical method for simultaneously quantifying 12 illicit drugs and metabolites(methamphetamine,amphetamine,morphine,codeine,6-monoacetylmorphine,benzoylecgonine,3,4-methylenedioxymethamphetamine,3,4-methylenedioxyamphetamine,cocaine,ketamine,norketamine,and methcathinone)and cotinine(COT)in wastewater samples.The analysis was performed by loading 2 m L of the sample onto an Oasis hydrophilic-lipophilic balance cartridge and using a cleanup step(5%methanol)to eliminate interference with a total run time of 13 min.The isotope-labeled internal standard method was used to quantify the target substances and correct for unavoidable losses and matrix effects during the on-line SPE process.Typical analytical characteristics used for method validation were sensitivity,linearity,precision,repeatability,recovery,and matrix effects.The limit of detection(LOD)and limit of quantification(LOQ)of each target were set at 0.20 ng/L and 0.50 ng/L,respectively.The linearity was between 0.5 ng/L and250 ng/L,except for that of COT.The intra-and inter-day precisions were<10.45%and 25.64%,respectively,and the relative recovery ranged from 83.74%to 162.26%.The method was used to analyze various wastewater samples from 33 cities in China,and the results were compared with the experimental results of identical samples analyzed using off-line SPE.The difference rate was between 19.91%and-20.44%,and the error range could be considered acceptable.These findings showed that on-line SPE is a suitable alternative to off-line SPE for the analysis of illicit drugs in samples.

UHPLC-MS/MS analysis of cAMP and cGMP in rat plasma as potential biomarkers of Yin-Yang disharmony in traditional Chinese medicine

Cyclic 3’,5’-adenosine monophosphate(cAMP)and cyclic 3’,5’-guanosine monophosphate(cGMP)are considered as potential biomarkers for Yin-Yang disharmony in traditional Chinese medicine.However,phosphodiesterase-mediated ex vivo degradation of these molecules in biological samples may result in their underestimation.In the present study,a ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)method was developed for determination of cAMP and cGMP in rat plasma,with special consideration of their stability ex vivo.Following precipitation of proteins from plasma samples with 0.4 M perchloric acid,the analytes were chromatographed on a Shimadzu Shimpack-XR-ODS II column with 2.5 mM ammonium acetate and methanol in gradient mode.The MS/MS detection was performed using multiple reaction monitoring in the positive electrospray ionization mode.The lower limit of quantification was 0.27 ng/mL for cAMP and 0.37 ng/mL for cGMP.The method was used to determine the plasma cAMP and cGMP levels in normal and Yin deficiency diabetic rats treated with or without Rehmannia glutinosa.The developed method may be useful for evaluating the regulatory effects of Chinese herbal medicine on the levels of cAMP and cGMP in the body.

On-line enrichment and determination of polycyclic aromatic hydrocarbons in atmospheric particulates using high performance liquid chromatography with fluorescence as detector

Seven polycyclic aromatic hydrocarbons （PAHs） in atmospheric particulates were determinated by high performance liquid chromatography （HPLC） with fluorescence detector using direction injection and an on-line enrichment trap column. The method simplified the sample pretreatment, saved time and increased the efficiency. With the on-line trap column, PAHs were separated availably even underground injecting 1.0 ml sample with relatively high column efficiency. The recoveries of the seven PAHs were from 85% to 120% for spiked atmospheric particulate sample. The limit of detection was 15.3-39.6 ng/L （S/N=3.3）. There were good linear correlations between the peak areas and concentrations of the seven kinds of PAHs in the range of 1-50 ng/ml with the correlation coefficients over 0.9970. Furthermore, it also indicated that the method is available to determine PAHs in atmospheric particulates well.

用在线样品预处理技术通过高效液相色谱测定血样中的药物

The flow diagram of this on-line sample pretreatment system is shown in Fig.1.

Schisandra lignans ameliorate nonalcoholic steatohepatitis by regulating aberrant metabolism of phosphatidylethanolamines

Nonalcoholic steatohepatitis(NASH)is a spectrum of chronic liver disease characterized by hepatic lipid metabolism disorder.Recent reports emphasized the contribution of triglyceride and diglyceride accumulation to NASH,while the other lipids associated with the NASH pathogenesis remained unexplored.The specific purpose of our study was to explore a novel pathogenesis and treatment strategy of NASH via profiling the metabolic characteristics of lipids.Herein,multi-omics techniques based on LC—Q-TOF/MS,LC—MS/MS and MS imaging were developed and used to screen the action targets related to NASH progress and treatment.A methionine and choline deficient(MCD)diet-induced mouse model of NASH was then constructed,and Schisandra lignans extract(SLE)was applied to alleviate hepatic damage by regulating the lipid metabolism-related enzymes CES2A and CYP4A14.Hepatic lipidomics indicated that MCD-diet led to aberrant accumulation of phosphatidylethanolamines(PEs),and SLE could significantly reduce the accumulation of intrahepatic PEs.Notably,exogenous PE(18:0/18:1)was proved to significantly aggravate the mitochondrial damage and hepatocyte apoptosis.Supplementing PE(18:0/18:1)also deteriorated the NASH progress by up regulating intrahepatic proinflammatory and fibrotic factors,while PE synthase inhibitor exerted a prominent hepatoprotective role.The current work provides new insights into the relationship between PE metabolism and the pathogenesis of NASH.

Single-cell RNA sequencing reveals a hierarchical transcriptional regulatory network of terpenoid biosynthesis in cotton secretory glandular cells

Plants can synthesize a wide range of terpenoids in response to various environmental cues.However,the specific regulatory mechanisms governing terpenoid biosynthesis at the cellular level remain largely elusive.In this study,we employed single-cell RNA sequencing to comprehensively characterize the transcriptome profile of cotton leaves and established a hierarchical transcriptional network regulating cellspecific terpenoid production.We observed substantial expression levels of genes associated with the biosynthesis of both volatile terpenes(such asβ-caryophyllene andβ-myrcene)and non-volatile gossypol-type terpenoids in secretory glandular cells.Moreover,two novel transcription factors,namely GoHSFA4a and GoNAC42,are identified to function downstream of the Gossypium PIGMENT GLAND FORMATION genes.Both transcription factors could directly regulate the expression of terpenoid biosynthetic genes in secretory glandular cells in response to developmental and environmental stimuli.For convenient retrieval of the single-cell RNA sequencing data generated in this study,we developed a user-friendly web server.Our findings not only offer valuable insights into the precise regulation of terpenoid biosynthesis genes in cotton leaves but also provide potential targets for cotton breeding endeavors.

Simultaneous quantification of ginsenoside Rg1 and its metabolites by HPLC–MS/MS:Rg1 excretion in rat bile, urine and feces

Ginsenoside Rg1(Rg1), the major effective component of ginseng, has been shown to have multiple bioactivities, but low oral bioavailability. The aim of this study was to develop a simple, sensitive and rapid high performance liquid chromatography–tandem mass spectrometry(LC–MS/MS) method, which could be used to validate and quantify the concentrations of Rg1 and its metabolites in Sprague-Dawley rat bile,urine, and feces after oral administration(25 mg/kg). Calibration curves offered satisfactory linearity(r40.995)within the determined ranges. Both intra-day and inter-day variances were less than 15%, and the accuracy was within 80–120%. The excretion recoveries of Rg1, ginsenoside Rh1(Rh1), and protopanaxatriol(Ppt) in bile,urine, and feces combined were all greater than 70%. The fecal excretion recoveries of Rg1, Rh1, and Ppt were40.11%, 22.19%, and 22.88%, respectively, whereas 6.88% of Rg1 and 0.09% of Rh1 were excreted in bile.Urinary excretion accounted for only 0.04% of Rg1. In conclusion, the observed excretion profiles for Rg1 and its metabolites after oral administration are helpful for understanding the poor oral bioavailability of Rg1 and will aid further investigations of Rg1 as a pharmacologically active component.

Effect of Ce on solute redistribution in liquid ahead of solid–liquid interface during solidification of Fe–4 wt.%Si alloy

The high efficiency of Ce addition in grain refinement ofδ-ferrite in a cast Fe–4 wt.%Si alloy was verified.In order to further understand the solute effect of Ce on the grain refinement of δ-ferrite,the conventional directional solidification technique,which enabled to freeze the solid–liquid interface to room temperature,was used to investigate the interfacial morphology and solute redistribution in the liquid at the front of the interface,together with thermodynamic calculation of the equilibrium partition coefficients of Ce and Si in Fe–4 wt.%Si–Ce system using the Equilib module and the FsStel database in FactSage software system.Metallographic examination using a laser scanning confocal microscope showed a transition of the solid–liquid interface from planar to cellular in the Fe–4 wt.%Si alloy after adding 0.0260 wt.%Ce during the directional solidification experiment.Further,electron probe microanalysis revealed an enhanced segregation of Si solute in the liquid at the front of the solid–liquid interface due to the Ce addition.This solute segregation is considered as the cause of planar to cellular interface transition,which resulted from the creation of constitutional supercooling zone.Thermodynamic calculation indicated that Ce also segregated at the solid–liquid interface and the Ce addition had negligible effect on the equilibrium partition coefficient of Si.It is reasonable to consider that the contribution of Ce to the grain refinement ofδ-ferrite in the cast Fe–4 wt.%Si alloy as a solute was marginal.