The human plasma protein binding of water soluble flavonoids in the peels of five spices of citrus fruits was studied by ultrafiltration combined with HPLC.The flavonoids were extracted separately by hot and cold wate...The human plasma protein binding of water soluble flavonoids in the peels of five spices of citrus fruits was studied by ultrafiltration combined with HPLC.The flavonoids were extracted separately by hot and cold water,and higher total flavonoid contents were detected in the former extracts than the latter ones.All the extracts show significant scavenging abilities to both ABTS and DPPH free radicals,which indicates the health benefits of the water extracts of citrus fruits peels.For DPPH radical,the IC50values of hot extract follow as Navel orange(NO)≈Mandarin orange(MO)< Lemon(LE)< Lo tangerine(LO)< Pomelo(PO),while the rank is NO< PO<LE≈MO<LO for ABTS radical.The HPLC results reveal that the kinds and contents of the flavonoids detected in the extracts are different among the species.MO extract has the most neohesperidin dihydrochalcone of 118.76 μmol/L and quercetrin of 211.81 μmol/L of which are much more than the rest extracts.Pomelo extract has the most plentiful flavonoids of naringin with a concentration of 303.28 μmol/L.The high contents of myricetrin and dihydromyricetin which both are potent free radical scavengers may explain the highest free radical scavenging activity of the NO extract.The plasma binding rates decrease with the increasing concentrations of flavonoids,and the flavonoids having plenty hydroxyl groups on both A ring and B ring of the molecular skeleton have relative higher plasma binding rates.In addition,the plasma binding rates of flavonoids with saturated C3-C4 bond decrease significantly with the increasing concentrations.展开更多
A simple and selective ultra performance liquid chromatography--electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） assay was developed for the determination of the human plasma protein binding of four...A simple and selective ultra performance liquid chromatography--electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） assay was developed for the determination of the human plasma protein binding of four bioactive ftavonoids （such as orientin and vitexin） in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C i s column with a mobile phase composed of 0.1% formic acid in acetonitrile and 0.1% aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITYTM TQD system was operated under the multiple reaction monitoring （MRM） mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations （r 〉 0.99） of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.展开更多
The stereoselective hydrolysis of esmolol in whole blood and in its separated components from rat,rabbit and human was investigated.Blood esterase activities were variable in different species in the order of rat>r...The stereoselective hydrolysis of esmolol in whole blood and in its separated components from rat,rabbit and human was investigated.Blood esterase activities were variable in different species in the order of rat>rabbit>human.Rat plasma showed the high esterase activity and had no stereoselectivity to enantiomers.Rabbit red blood cell（RBC） membrane,RBC cytosol and plasma all hydrolyzed esmolol but with different esterase activity,whereas the hydrolysis in RBC membrane and cytosol showed significant stereoselectivity towards R-（+）-esmolol.Esterase in RBC cytosol from human blood mainly contributed to the esmolol hydrolysis,which was demonstrated with no stereoselctivity.Esterase in human plasma showed a low activity,but a remarkable stereoselectivity with R-（+）-esmolol.In addition,the protein concentration affected the hydrolysis behavior of esmolol in RBC suspension.Protein binding of esmolol enantiomers in human plasma,human serum albumin（HSA） and α;-acid glycoprotein（AGP） revealed that there was a significant difference in bound fractions between two enantiomers,especially for AGP.Our results indicated that the stereoselective protein binding might play a role in the different hydrolysis rates of esmolol enantiomers in human plasma.展开更多
In this paper a new continuous variable called core-ratio is defined to describe the probability for a residue to be in a binding site, thereby replacing the previous binary description of the interface residue using ...In this paper a new continuous variable called core-ratio is defined to describe the probability for a residue to be in a binding site, thereby replacing the previous binary description of the interface residue using 0 and 1. So we can use the support vector machine regression method to fit the core-ratio value and predict the protein binding sites. We also design a new group of physical and chemical descriptors to characterize the binding sites. The new descriptors are more effective, with an averaging procedure used. Our test shows that much better prediction results can be obtained by the support vector regression （SVR） method than by the support vector classification method.展开更多
High density lipoprotein binding protein （HBP） plays an important role in lipid metabolism of animals. Lipids are indispensable energy materials for fi- shes, especially for carnivorous fishes with low utilization e...High density lipoprotein binding protein （HBP） plays an important role in lipid metabolism of animals. Lipids are indispensable energy materials for fi- shes, especially for carnivorous fishes with low utilization efficiency of carbohydrates. The single nucleotide polymorphism （SNP） of HBP gene may affect the fat metabolism, thereby exerting an effect on the growth traits of largemouth bass （Micropterus salmoides）. Investigating the correlations between SNP and growth traits can provide candidate markers for molecular marker-assisted selection. In this study, partial genomic fraganents of HBP gene （ GenBank accession number： KF652241 ） were amplified based on the sequences of an available contig in the EST-SNP database of largemouth bass. Three SNP mutation loci were identified in the 3＇ non-ceding region of HBP gene by direct sequencing, including H1 （G ＋ 2782T）, 142 （T ＋ 2817C） and H3 （G ＋ 2857A）. Three SNP loci of 165 randomly selected largemouth bass individuals were detected and genotyped by SnaPshot assay. Genetic structure was analyzed by POPGENE32 software. By using spssl7.0 software, a general linear model （GLM） was established for correlation analysis between different genotypes at SNP loci of HBP gene and various growth traits. The results showed that three SNP loci were in Hardy-Weinberg equilibrium state. To be specific, loci H1 and H2 formed two haplotypes （ A and B）, and three geno- types （AA, AB, and BB） were observed; loci H1, H2 and H3 formed six diplotypes （DI, I）2, D3, D4, D5 and D6）. According to the correlations between dif- ferent genotypes and various growth traits, the body weight and total length of largemouth bass individuals with genotype BB were significantly higher than those of individuals with genotype AB （ P 〈 0.05 ） ; the body weight and total length of largemouth bass individuals with diplotype D6 were significantly higher than those of individuals with diplotype D4 （P 〈0.05）. In this study, SNP markers correlated with growth traits were obtained in the 3＇ non-coding region ofHBP gene in large-mouth bass, which could be used as candidate genetic markers for subsequent molecular marker-assisted selection breeding of largemouth bass.展开更多
TM-2 known as a potential antitumor drug is a novel semi-synthetic taxane derivative. As drug-protein interactions contribute to insights into pharmacokinetic and pharmacodynamic properties, we eluci- dated the bindin...TM-2 known as a potential antitumor drug is a novel semi-synthetic taxane derivative. As drug-protein interactions contribute to insights into pharmacokinetic and pharmacodynamic properties, we eluci- dated the binding of TM-2 to plasma protein. In this study, a simple, rapid and reliable method was developed and validated employing equilibrium dialysis for the separation of bound and unbound drugs and ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for the quantitation. Protein binding reached equilibrium within 24 h of incubation at 37 ℃. After liquid-liquid extraction with methyl tert-butyl ether, the samples were separated on Thermo Syncronis UPLC C18 （2.1 mm× 50 mm, 1.7 μm）, and acquisition of mass spectrometric data was performed in multiple re- action monitoring （MRM） mode via positive electrospray ionization. The assay was linear over the concentration rang of 5-2000 nglmL The intra- and inter-day precisions were 0.1%-14.8%, and the accuracy was from -6.4% to Z0%. This assay has been successfully applied to a protein binding study of TM-2 in rat, human and beagle dog plasma. TM-2 showed high protein binding of 81.4% ± 6.5% （rat）, 87.9% ± 3.6% （human） and 79.4% ± 4.0% （beagle dog）. The results revealed that there was an insignificant difference among the three species.展开更多
Background Post-weaning diarrhea(PWD)in piglets,often caused by F4^(+)enterotoxigenic Escherichia coli(ETEC),poses significant challenges in pig production.Traditional solutions like antibiotics and zinc oxide face in...Background Post-weaning diarrhea(PWD)in piglets,often caused by F4^(+)enterotoxigenic Escherichia coli(ETEC),poses significant challenges in pig production.Traditional solutions like antibiotics and zinc oxide face increasing restrictions due to growing concerns over antibiotic resistance and environmental sustainability.This study investigates the application of bivalent heavy chain variable domain(V_(H)H)constructs(BL1.2 and BL2.2)targeting ETEC virulence factors,administered in feed to mitigate ETEC-induced PWD in weaned piglets.Results The supplementation of BL1.2 and BL2.2 in both mash and pelleted feed significantly reduced the diarrhea incidence and fecal shedding of F4^(+)ETEC in challenged piglets.Pelleted feed containing V_(H)H constructs helped to preserve gut barrier integrity by maintaining levels of the tight junction protein occludin in the small intestine.Additionally,the constructs maintained blood granulocyte counts at a similar level to the non-challenged control group,including neutrophils,and ameliorated the acute phase protein response after challenge.Notably,even at low feed intake immediately after weaning,V_(H)H constructs helped maintain piglet health by mitigating ETEC-induced inflammation and the resulting diarrhea.Conclusions Our findings demonstrated that using V_(H)H constructs as feed additives could serve as an effective strategy to help manage ETEC-associated PWD,by reducing F4^(+)ETEC gut colonization and supporting gut barrier function of weaned piglets.The high stability of these V_(H)H constructs supports their incorporation into industrial feed manufacturing processes,offering a more sustainable preventive strategy compared to traditional antimicrobial interventions,which could contribute to sustainable farming practices.展开更多
Magnolol,a compound extracted from Magnolia officinalis,demonstrates potential efficacy in addressing metabolic dysfunction and cardiovascular diseases.Its biological activities encompass anti-inflammatory,antioxidant...Magnolol,a compound extracted from Magnolia officinalis,demonstrates potential efficacy in addressing metabolic dysfunction and cardiovascular diseases.Its biological activities encompass anti-inflammatory,antioxidant,anticoagulant,and anti-diabetic effects.Growth/differentiation factor-15(GDF-15),a member of the transforming growth factorβsuperfamily,is considered a potential therapeutic target for metabolic disorders.This study investigated the impact of magnolol on GDF-15 production and its underlying mechanism.The research examined the pharmacological effect of magnolol on GDF-15 expression in vitro and in vivo,and determined the involvement of endoplasmic reticulum(ER)stress signaling in this process.Luciferase reporter assays,chromatin immunoprecipitation,and in vitro DNA binding assays were employed to examine the regulation of GDF-15 by activating transcription factor 4(ATF4),CCAAT enhancer binding proteinγ(CEBPG),and CCCTC-binding factor(CTCF).The study also investigated the effect of magnolol and ATF4 on the activity of a putative enhancer located in the intron of the GDF-15 gene,as well as the influence of single nucleotide polymorphisms(SNPs)on magnolol and ATF4-induced transcription activity.Results demonstrated that magnolol triggers GDF-15 production in endothelial cells(ECs),hepatoma cell line G2(HepG2)and hepatoma cell line 3B(Hep3B)cell lines,and primary mouse hepatocytes.The cooperative binding of ATF4 and CEBPG upstream of the GDF-15 gene or the E1944285 enhancer located in the intron led to full-power transcription of the GDF-15 gene.SNP alleles were found to impact the magnolol and ATF4-induced transcription activity of GDF-15.In high-fat diet ApoE^(-/-)mice,administration of magnolol induced GDF-15 production and partially suppressed appetite through GDF-15.These findings suggest that magnolol regulates GDF-15 expression through priming of promoter and enhancer activity,indicating its potential as a drug for the treatment of metabolic disorders.展开更多
BACKGROUND The Mac-2 binding protein glycosylated isomer(M2BPGi)is a serum marker for fibrosis that correlates with the fibrosis stages in various liver diseases.AIM To examine the M2BPGi’s threshold for staging fibr...BACKGROUND The Mac-2 binding protein glycosylated isomer(M2BPGi)is a serum marker for fibrosis that correlates with the fibrosis stages in various liver diseases.AIM To examine the M2BPGi’s threshold for staging fibrosis in patients with chronic hepatitis B(CHB),and its changes during treatment.METHODS This was a prospective,longitudinal study.A total of 348 eligible patients were recruited from the Hepatology Department,Medic Medical Center between March 2020 and December 2023.Liver enzyme tests,platelet counts,M2BPGi levels,and FibroScan were conducted at baseline and at 3-month intervals until six months post-treatment.Correlation plots of M2BPGi,FibroScan,and the other parameters were generated.Receiver operating characteristic curves were constructed for M2BPGi and the other parameters to evaluate their performance.RESULTS M2BPGi levels correlated well with FibroScan results and increased as the fibrosis stage advanced.The median M2BPGi levels at the different stages of fibrosis showed statistically significant differences.The cut-off values of M2BPGi for diagnosing significant fibrosis(F≥2),advanced fibrosis(F3),and cirrhosis(F4)were determined to be 1.08,1.4,and 1.52,respectively.In the context of fibrosis regression in CHB patients during the first 6-month of treatment,M2BPGi levels appeared to decrease before this pattern occurred in the FibroScan results.CONCLUSION M2BPGi levels were strongly correlated with FibroScan.M2BPGi can be used to assess liver fibrosis,and to serve as a tool for monitoring fibrosis regression in CHB patients undergoing treatment.展开更多
BACKGROUND Cirrhosis is a progressive condition characterized by fibrosis that can lead to severe complications and increased mortality.The mac-2 binding protein glyco-sylation isomer(M2BPGi)is a prominent biomarker f...BACKGROUND Cirrhosis is a progressive condition characterized by fibrosis that can lead to severe complications and increased mortality.The mac-2 binding protein glyco-sylation isomer(M2BPGi)is a prominent biomarker for predicting hepatocellular carcinoma(HCC)and cirrhosis-induced esophageal varices(EV).AIM To investigate thresholds of M2BPGi associated with HCC,EV,and decomp-ensation in patients with cirrhosis.METHODS This was a prospective study.A total of 153 patients with cirrhosis who met the inclusion criteria were enrolled.The patients were diagnosed with HCC and EV according to the Baveno VII and European Association for the Study of the Liver guidelines.Baseline serum M2BPGi levels were assessed along with other routine tests.The data analysis aimed to determine the cutoff values of M2BPGi for pre-dicting EV and HCC.RESULTS In the study 85.6%of patients were Child-Pugh B and C.M2BPGi mean cutoff index was 7.1±3.7,showing no significant etiological differences.However,M2BPGi levels varied significantly among Child-Pugh classes,EV classifications,and between patients with and without HCC(P<0.01).M2BPGi cutoff values for predicting HCC,EV,and decompensated cirrhosis were 6.50,6.64,and 5.25,respectively.Mul-tivariate analysis confirmed M2BPGi as an independent risk factor for EV[adjusted odds ratio(aOR):1.3,95%CI:1.08-1.64]and liver decompensation(aOR:2.11,95%CI:1.37-3.83).Area under the curve of M2BPGi for HCC differ-entiation was 0.71.An algorithm combining alpha-fetoprotein(AFP)and M2BPGi detected 26 of 28 HCC cases with 98.04%accuracy vs 10 cases by AFP alone.CONCLUSION Serum M2BPGi predicted cirrhosis complications,including decompensation and varices,especially in HCC.Combined with AFP,it enhanced HCC detection.Future liver biopsy studies are needed for tissue confirmation.展开更多
BACKGROUND Mac-2 binding protein glycosylation isomer(M2BPGi)serves as a marker of activated hepatic stellate cells and as such holds potential as a biomarker for liver fibrosis.In Viet Nam,metabolic dysfunction-assoc...BACKGROUND Mac-2 binding protein glycosylation isomer(M2BPGi)serves as a marker of activated hepatic stellate cells and as such holds potential as a biomarker for liver fibrosis.In Viet Nam,metabolic dysfunction-associated steatotic liver disease(MASLD)is rising in prevalence and there is an urgent need for better clinical management,particularly in early detection methods that will improve overall prognosis.AIM To examine M2BPGi cut-off values for staging liver fibrosis in patients with MASLD and risk factors associated with disease progression.METHODS A total of 301 individuals with ultrasound-confirmed or FibroScan-confirmed diagnosis of fatty liver were enrolled in the study.The participants were stratified according to fibrosis stage,measured via magnetic resonance elastography.M2-BPGi,Fibrosis-4(FIB-4)Index score,and routine parameters of liver function were assessed to statistically investigate the correlation of M2BPGi levels in various fibrosis stages and to identify risk factors associated with fibrosis severity.RESULTS M2BPGi levels positively correlated with fibrosis stages,with cut-off indexes of 0.57 for F0-1,0.68 for F2-3,and 0.78 for F4.M2BPGi levels in the F0-1 group were significantly different from those in both the F2-3 group(P=0.038)and the F4 group(P=0.0051);the F2-3 and F4 groups did not show a significant difference(P=0.39).Females exhibited significantly higher M2BPGi levels than males for all fibrosis stages,particularly in the F2-3 group(P=0.01)and F4 group(P=0.0006).In the F4(cirrhosis)group,individuals with diabetes had significantly higher M2BPGi levels than those without.M2BPGi,hemoglobin A1c,and FIB-4 score were identified as independent risk factors for greater fibrosis and cirrhosis.CONCLUSION M2BPGi levels varied significantly throughout fibrosis progression,from early MASLD to cirrhosis,with sex correlation.M2BPGi holds promise as an early biomarker for fibrosis characterization in MASLD adult patient populations.展开更多
Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome,with implications for microbial pathogenesis and host defense.Among these,transfer RNA-de...Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome,with implications for microbial pathogenesis and host defense.Among these,transfer RNA-derived small RNAs(tsRNAs)have garnered attention for their roles in modulating microbial behavior.However,the bacterial factors mediating tsRNA interaction and functionality remain poorly understood.In this study,using RNA affinity pull-down assay in combination with mass spectrometry,we identified a putative membrane-bound protein,annotated as P-type ATPase transporter(PtaT)in Fusobacterium nucleatum(Fn),which binds Fn-targeting tsRNAs in a sequence-specific manner.Through targeted mutagenesis and phenotypic characterization,we showed that in both the Fn type strain and a clinical tumor isolate,deletion of ptaT led to reduced tsRNA intake and enhanced resistance to tsRNA-induced growth inhibition.Global RNA sequencing and label-free Raman spectroscopy revealed the phenotypic differences between Fn wild type and PtaT-deficient mutant,highlighting the functional significance of PtaT in purine and pyrimidine metabolism.Furthermore,AlphaFold 3 prediction provides evidence supporting the specific binding between PtaT and Fn-targeting tsRNA.By uncovering the first RNA-binding protein in Fn implicated in growth modulation through interactions with host-derived small RNAs(sRNAs),our study offers new insights into sRNA-mediated host-pathogen interplay within the context of microbiome-host interactions.展开更多
BACKGROUND The exact mechanisms underlying diabetic nephropathy(DN)remain incompletely elucidated,prompting researchers to explore new perspectives and identify novel intervention targets in this field.AIM To explore ...BACKGROUND The exact mechanisms underlying diabetic nephropathy(DN)remain incompletely elucidated,prompting researchers to explore new perspectives and identify novel intervention targets in this field.AIM To explore the role and underlying mechanisms of farnesoid X receptor(FXR)in the development of DN by regulating endoplasmic reticulum stress(ERS)molecular chaperone binding immunoglobulin protein(BiP)expression.METHODS Bioinformatics analyses identified potential FXR-binding elements in the BiP promoter.Dual-luciferase and chromatin immunoprecipitation(ChIP)assays confirmed FXR-BiP binding sites.In vitro studies used SV40 MES 13 cells under varying glucose conditions and treatments with FXR modulators[obeticholic acid(INT-747)and guggulsterones]or BiP small interfering RNA.The expression of BiP and ERS-related proteins[protein kinase R-like endoplasmic reticulum kinase(PERK),inositol-requiring enzyme 1(IRE1),activating transcription factor 6(ATF6)]was assessed alongside cell proliferation and extracellular matrix(ECM)synthesis.In vivo studies in DN mice(db/db)examined the effects of FXR activation on renal function and morphology.RESULTS FXR bound to the target sequence in the BiP promoter region,enhancing transcriptional activity,as confirmed by ChIP experiments.FXR expression decreased in SV40 MES 13 cells stimulated with high glucose and in renal tissues of DN mice compared with control.Treatment of SV40 MES 13 cells with the FXR agonist INT-747 significantly increased intracellular BiP expression,whereas silencing the FXR gene led to the downregulation of BiP levels.In vivo administration of INT-747 significantly elevated BiP levels in renal tissues,improved renal function and fibrosis in DN mice,while inhibiting the expression of ERS-related signaling proteins PERK,IRE1,and ATF6.CONCLUSION FXR promotes BiP expression by binding to its promoter,suppressing ERS pathways,and reducing mesangial cell proliferation and ECM synthesis.These findings highlight FXR as a potential therapeutic target for diabetic glomerulosclerosis.展开更多
BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the rol...BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the role of MLKL’s adenosine triphosphate(ATP)-binding pocket in facilitating necroptosis-independent pathways that may contribute to liver disease progression.By focusing on this mechanism,we seek to identify potential therapeutic targets that can modulate MLKL activity,offering new strategies for the prevention and treatment of liver-related pathologies.AIM To investigate the possibility of using the ATP-binding pocket-associated,necro-ptosis-independent MLKL pathway as a target for liver diseases.METHODS Cell death following necroptosis stimuli was evaluated using cell proliferation assays,flow cytometry,and electron microscopy in various cells.The human liver organoid system was used to evaluate whether the MLKL ATP pocket-binding inhibitor could attenuate inflammation.Additionally,alcoholic and non-alcoholic fatty liver diseases animal models were used to determine whether MLKL ATP pocket inhibitors could attenuate liver injury.RESULTS While an MLKL ATP pocket-binding inhibitor did not prevent necroptosis-induced cell death in RAW 264.7 cells,it did reduce the necroptosis-led expression of CXCL2,ICAM,and VCAM.Notably,MLKL ATP pocket inhibitor diminishes the expression of CXCL2,ICAM,and VCAM by inhibiting the IκB kinase and nuclear factor kappa-B pathways without inducing necroptosis-induced cell death in two-dimensional cell culture as well as the human-derived liver organoid system.Although MLKL ATP-binding inhibitor was ineffective in non-alcoholic fatty liver disease animal models,MLKL ATP-binding inhibitor attenuated hepatic inflammation in the alcoholic liver disease model.CONCLUSION MLKL ATP pocket-binding inhibitor exerted anti-inflammatory effects through the necroptosis-independent MLKL pathway in an animal model of alcoholic liver disease.展开更多
Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBI...Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBIRC6)on non-small cell lung cancer(NSCLC)progression.Methods:The NSCLC and adjacent non-tumor tissues were collected at Shanghai Ninth People's Hospital.Quantitative real-time polymerase chain reaction was conducted for assessing the levels of circBIRC6,amyloid beta precursor protein binding protein 2(APPBP2)messenger RNA(mRNA),baculoviral IAP repeat containing 6 mRNA(BIRC6),and microRNA-217(miR-217).Western blot assay was adopted for measuring the protein levels of APPBP2,E-cadherin,N-cadherin,and vimentin.Colony formation assay,transwell assay,and flow cytometry analysis were utilized for evaluating cell colony formation,metastasis,and apoptosis.Dualluciferase reporter assay and RNA immunoprecipitation assay were carried out to determine the interaction between miR-217 and circBIRC6 and APPBP2 in NSCLC tissues.The murine xenograft model assay was used to investigate the function of circBIRC6 in tumor formation in vivo.Differences were analyzed via Student's t test or one-way analysis of variance.Pearson's correlation coefficient analysis was used to analyze linear correlation.Results:CircBIRC6 was overexpressed in NSCLC tissues and cells.Knockdown of circBIRC6 repressed the colony formation and metastasis and facilitated apoptosis of NSCLC cells in vitro and restrained tumorigenesis in vivo.Mechanically,circBIRC6 functioned as miR-217 sponge to promote APPBP2 expression in NSCLC cells.MiR-217 inhibition rescued circBIRC6 knockdown-mediated effects on NSCLC cell colony formation,metastasis,and apoptosis.Overexpression of miR-217 inhibited the malignant phenotypes of NSCLC cells,while the effects were abrogated by elevating APPBP2.Conclusion:CircBIRC6 aggravated NSCLC cell progression by elevating APPBP2 via sponging miR-217,which might provide a fresh perspective on NSCLC therapy.展开更多
[Objective] The research aimed to find the extracellular binding proteins of CR4.[Method] The extracellular domain of OsCR4 was as the bait protein,and the yeast two-hybrid was used to screen cDNA library of seedling ...[Objective] The research aimed to find the extracellular binding proteins of CR4.[Method] The extracellular domain of OsCR4 was as the bait protein,and the yeast two-hybrid was used to screen cDNA library of seedling which was cultivated 14 d.[Result] A lot of proteins which included a peroxide B（D26484）,a methionine thioredoxin reductase（ABF96078）and an unknown function protein were gained.[Conclusion] It provided the theory basis for studying the signal transduction mechanism of CR4.展开更多
The molecular mechanism of how hepatocytes maintain cholesterol homeostasis has become much more transparent with the discovery of sterol regulatory element binding proteins (SREBPs) in recent years. These membrane pr...The molecular mechanism of how hepatocytes maintain cholesterol homeostasis has become much more transparent with the discovery of sterol regulatory element binding proteins (SREBPs) in recent years. These membrane proteins aremembers of the basic helix-loop-helix-leucine zipper (bHLHZip) family of transcription factors. They activate the expression of at least 30 genes involved in the synthesis of cholesterol and lipids. SREBPs are synthesized as precursor proteins in the endoplasmic reticulum (ER), where they form a complex with another protein, SREBP cleavage activating protein (SCAP). The SCAP molecule contains a sterol sensory domain. In the presence of high cellular sterol concentrations SCAP confines SREBP to the ER. With low cellular concentrations, SCAP escorts SREBP to activation in the Golgi. There, SREBP undergoes two proteolytic cleavage steps to release the mature, biologically active transcription factor, nuclear SREBP (nSREBP). nSREBP translocates to the nucleus and binds to sterol response elements (SRE) in the promoter/enhancer regions of target genes. Additional transcription factors are required to activate transcription of these genes. Three different SREBPs are known, SREBPs-1a, -1c and -2. SREBP-1a and -1c are isoforms produced from a single gene by alternate splicing. SREBP-2 is encoded by a different gene and does not display any isoforms. It appears that SREBPs alone, in the sequence described above, can exert complete control over cholesterol synthesis, whereas many additional factors (hormones, cytokines, etc.) are required for complete control of lipid metabolism. Medicinal manipulation of the SREBP/SCAP system is expected to prove highly beneficial in the management of cholesterol-related disease.展开更多
The association of retinol binding protein 4 （RBP4） with atherosclerosis of the carotid artery in type 2 diabetes mellitus （T2DM） remains undefined. We aimed to investigate the correlation of RBP4 expression with ...The association of retinol binding protein 4 （RBP4） with atherosclerosis of the carotid artery in type 2 diabetes mellitus （T2DM） remains undefined. We aimed to investigate the correlation of RBP4 expression with atherosclerosis of the carotid artery in T2DM. A total of 1,076 subjects were investigated for intima-media thickness of the bilateral common carotid arteries, and they were divided into three groups： in group Ⅰ, patients had normal neck vascular ultra- sound, in group Ⅱ, intimal carotid artery media thickness was equal to or more than 1 mm, and in group Ⅲ, carotid artery plaque was present. Height, weight, blood pressure （BP）, fasting plasma glucose （FPG）, hemoglobin Alc （HbA1c）, total cholesterol （TC）, triglyceride （TG）, low-density lipoprotein cholesterol （LDL-C）, high-density lipopro- tein cholesterol （HDL-C）, apolipoprotein A-1 （apoA-1）, apolipoprotein B （apoB） and lipoprotein （a） [Lp（a）] were determined by routine laboratory methods. RBP4 and high sensitivity C reactive protein （HsCRP） were measured by an enzyme-linked immuno-sorbent assay, and insulin concentration was measured by an electrochemiluminescence sandwich immunoassay. Duration of diabetes, waist and BP, FPG, HbAlc, TG, TC, LDL-C, APOB, Lp（a）, HsCRP, RBP4 and homeostasis model assessment insulin resistance index （HOMA-IR） were significantly lower in group I than in the other two groups （P〈0.01, P〈0.01）. Plasma levels of HbAlc, RBP4, LDL-C, TC, HOMA-IR, HsCRP and Lp（a）, waist and BP were significantly increased in group III than in group II （P〈0.01）. Multivariate logistic regression analysis showed that there were seven factors associated with the occurrence of carotid artery atherosclero- sis and its risks in descending order were： high LDL-C, high waist, high HsCRP, duration of diabetes, high HOMA-IR, HbAlc and high RBP4. Our finding supported that RBP4 was positively correlated with carotid atherosclerosis in patients with T2DM and could be used as an early predictor of cardiovascular disease.展开更多
Protein binding is essential to the transport,decay and regulation of almost all RNA molecules.However,the structural preference of protein binding on RNAs and their cellular functions and dynamics upon changing envir...Protein binding is essential to the transport,decay and regulation of almost all RNA molecules.However,the structural preference of protein binding on RNAs and their cellular functions and dynamics upon changing environmental conditions are poorly understood.Here,we integrated various high-throughput data and introduced a computational framework to describe the global interactions between RNA binding proteins(RBPs)and structured RNAs in yeast at single-nucleotide resolution.We found that on average,in terms of percent total lengths,~15%of mRNA untranslated regions(UTRs),~37%of canonical non-coding RNAs(ncRNAs)and^11%of long ncRNAs(lncRNAs)are bound by proteins.The RBP binding sites,in general,tend to occur at single-stranded loops,with evolutionarily conserved signatures,and often facilitate a specific RNA structure conformation in vivo.We found that four nucleotide modifications of tRNA are significantly associated with RBP binding.We also identified various structural motifs bound by RBPs in the UTRs of mRNAs,associated with localization,degradation and stress responses.Moreover,we identified>200 novel lncRNAs bound by RBPs,and about half of them contain conserved secondary structures.We present the first ensemble pattern of RBP binding sites in the structured non-coding regions of a eukaryotic genome,emphasizing their structural context and cellular functions.展开更多
The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,...The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,encoding a protein with 147 amino acids,namely HarmOBP5.HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography.SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics.Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles,including （E）-β-farnesene,ethyl butyrate,ethyl heptanoate,and acetic acid 2-methylbutyl ester.Based on three-dimensional （3D） model of AaegOBP1 from Aedes aegypti,a 3D model of HarmOBP5 was predicted.The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H.armigera.This study provides clues for better understanding physiological functions of OBPs in H.armigera and other insects.展开更多
基金Project(21176263) supported by the National Natural Science Foundation of China
文摘The human plasma protein binding of water soluble flavonoids in the peels of five spices of citrus fruits was studied by ultrafiltration combined with HPLC.The flavonoids were extracted separately by hot and cold water,and higher total flavonoid contents were detected in the former extracts than the latter ones.All the extracts show significant scavenging abilities to both ABTS and DPPH free radicals,which indicates the health benefits of the water extracts of citrus fruits peels.For DPPH radical,the IC50values of hot extract follow as Navel orange(NO)≈Mandarin orange(MO)< Lemon(LE)< Lo tangerine(LO)< Pomelo(PO),while the rank is NO< PO<LE≈MO<LO for ABTS radical.The HPLC results reveal that the kinds and contents of the flavonoids detected in the extracts are different among the species.MO extract has the most neohesperidin dihydrochalcone of 118.76 μmol/L and quercetrin of 211.81 μmol/L of which are much more than the rest extracts.Pomelo extract has the most plentiful flavonoids of naringin with a concentration of 303.28 μmol/L.The high contents of myricetrin and dihydromyricetin which both are potent free radical scavengers may explain the highest free radical scavenging activity of the NO extract.The plasma binding rates decrease with the increasing concentrations of flavonoids,and the flavonoids having plenty hydroxyl groups on both A ring and B ring of the molecular skeleton have relative higher plasma binding rates.In addition,the plasma binding rates of flavonoids with saturated C3-C4 bond decrease significantly with the increasing concentrations.
基金This work was supported by the National Science Foundation of China （No. 30860366） Guizhou Province Municipal Science and Technology Project （No. 2007-6010）.
文摘A simple and selective ultra performance liquid chromatography--electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） assay was developed for the determination of the human plasma protein binding of four bioactive ftavonoids （such as orientin and vitexin） in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C i s column with a mobile phase composed of 0.1% formic acid in acetonitrile and 0.1% aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITYTM TQD system was operated under the multiple reaction monitoring （MRM） mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations （r 〉 0.99） of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.
基金supported by National Major Projects of Ministry Science and Technology of China(2011CB710800,2012ZX09506001-004)Zhejiang Education Department(Y200909571)
文摘The stereoselective hydrolysis of esmolol in whole blood and in its separated components from rat,rabbit and human was investigated.Blood esterase activities were variable in different species in the order of rat>rabbit>human.Rat plasma showed the high esterase activity and had no stereoselectivity to enantiomers.Rabbit red blood cell（RBC） membrane,RBC cytosol and plasma all hydrolyzed esmolol but with different esterase activity,whereas the hydrolysis in RBC membrane and cytosol showed significant stereoselectivity towards R-（+）-esmolol.Esterase in RBC cytosol from human blood mainly contributed to the esmolol hydrolysis,which was demonstrated with no stereoselctivity.Esterase in human plasma showed a low activity,but a remarkable stereoselectivity with R-（+）-esmolol.In addition,the protein concentration affected the hydrolysis behavior of esmolol in RBC suspension.Protein binding of esmolol enantiomers in human plasma,human serum albumin（HSA） and α;-acid glycoprotein（AGP） revealed that there was a significant difference in bound fractions between two enantiomers,especially for AGP.Our results indicated that the stereoselective protein binding might play a role in the different hydrolysis rates of esmolol enantiomers in human plasma.
基金Project supported by the National Natural Science Foundation of China (Grant Nos. 10674172 and 10874229)
文摘In this paper a new continuous variable called core-ratio is defined to describe the probability for a residue to be in a binding site, thereby replacing the previous binary description of the interface residue using 0 and 1. So we can use the support vector machine regression method to fit the core-ratio value and predict the protein binding sites. We also design a new group of physical and chemical descriptors to characterize the binding sites. The new descriptors are more effective, with an averaging procedure used. Our test shows that much better prediction results can be obtained by the support vector regression （SVR） method than by the support vector classification method.
基金Supported by National Natural Science Foundation of China(No.31201985)National Key Technology Support Program of China(2012BAD26B03)"948"Key Program of the Ministry of Agriculture of China(2011-G12)
文摘High density lipoprotein binding protein （HBP） plays an important role in lipid metabolism of animals. Lipids are indispensable energy materials for fi- shes, especially for carnivorous fishes with low utilization efficiency of carbohydrates. The single nucleotide polymorphism （SNP） of HBP gene may affect the fat metabolism, thereby exerting an effect on the growth traits of largemouth bass （Micropterus salmoides）. Investigating the correlations between SNP and growth traits can provide candidate markers for molecular marker-assisted selection. In this study, partial genomic fraganents of HBP gene （ GenBank accession number： KF652241 ） were amplified based on the sequences of an available contig in the EST-SNP database of largemouth bass. Three SNP mutation loci were identified in the 3＇ non-ceding region of HBP gene by direct sequencing, including H1 （G ＋ 2782T）, 142 （T ＋ 2817C） and H3 （G ＋ 2857A）. Three SNP loci of 165 randomly selected largemouth bass individuals were detected and genotyped by SnaPshot assay. Genetic structure was analyzed by POPGENE32 software. By using spssl7.0 software, a general linear model （GLM） was established for correlation analysis between different genotypes at SNP loci of HBP gene and various growth traits. The results showed that three SNP loci were in Hardy-Weinberg equilibrium state. To be specific, loci H1 and H2 formed two haplotypes （ A and B）, and three geno- types （AA, AB, and BB） were observed; loci H1, H2 and H3 formed six diplotypes （DI, I）2, D3, D4, D5 and D6）. According to the correlations between dif- ferent genotypes and various growth traits, the body weight and total length of largemouth bass individuals with genotype BB were significantly higher than those of individuals with genotype AB （ P 〈 0.05 ） ; the body weight and total length of largemouth bass individuals with diplotype D6 were significantly higher than those of individuals with diplotype D4 （P 〈0.05）. In this study, SNP markers correlated with growth traits were obtained in the 3＇ non-coding region ofHBP gene in large-mouth bass, which could be used as candidate genetic markers for subsequent molecular marker-assisted selection breeding of largemouth bass.
基金partly supported by the National High Technology Research and Development Program of China(No.2012AA020305)
文摘TM-2 known as a potential antitumor drug is a novel semi-synthetic taxane derivative. As drug-protein interactions contribute to insights into pharmacokinetic and pharmacodynamic properties, we eluci- dated the binding of TM-2 to plasma protein. In this study, a simple, rapid and reliable method was developed and validated employing equilibrium dialysis for the separation of bound and unbound drugs and ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for the quantitation. Protein binding reached equilibrium within 24 h of incubation at 37 ℃. After liquid-liquid extraction with methyl tert-butyl ether, the samples were separated on Thermo Syncronis UPLC C18 （2.1 mm× 50 mm, 1.7 μm）, and acquisition of mass spectrometric data was performed in multiple re- action monitoring （MRM） mode via positive electrospray ionization. The assay was linear over the concentration rang of 5-2000 nglmL The intra- and inter-day precisions were 0.1%-14.8%, and the accuracy was from -6.4% to Z0%. This assay has been successfully applied to a protein binding study of TM-2 in rat, human and beagle dog plasma. TM-2 showed high protein binding of 81.4% ± 6.5% （rat）, 87.9% ± 3.6% （human） and 79.4% ± 4.0% （beagle dog）. The results revealed that there was an insignificant difference among the three species.
基金financially supported by the Green Development and Demonstration Programme(GUDP)(case number 34009-19-1585)。
文摘Background Post-weaning diarrhea(PWD)in piglets,often caused by F4^(+)enterotoxigenic Escherichia coli(ETEC),poses significant challenges in pig production.Traditional solutions like antibiotics and zinc oxide face increasing restrictions due to growing concerns over antibiotic resistance and environmental sustainability.This study investigates the application of bivalent heavy chain variable domain(V_(H)H)constructs(BL1.2 and BL2.2)targeting ETEC virulence factors,administered in feed to mitigate ETEC-induced PWD in weaned piglets.Results The supplementation of BL1.2 and BL2.2 in both mash and pelleted feed significantly reduced the diarrhea incidence and fecal shedding of F4^(+)ETEC in challenged piglets.Pelleted feed containing V_(H)H constructs helped to preserve gut barrier integrity by maintaining levels of the tight junction protein occludin in the small intestine.Additionally,the constructs maintained blood granulocyte counts at a similar level to the non-challenged control group,including neutrophils,and ameliorated the acute phase protein response after challenge.Notably,even at low feed intake immediately after weaning,V_(H)H constructs helped maintain piglet health by mitigating ETEC-induced inflammation and the resulting diarrhea.Conclusions Our findings demonstrated that using V_(H)H constructs as feed additives could serve as an effective strategy to help manage ETEC-associated PWD,by reducing F4^(+)ETEC gut colonization and supporting gut barrier function of weaned piglets.The high stability of these V_(H)H constructs supports their incorporation into industrial feed manufacturing processes,offering a more sustainable preventive strategy compared to traditional antimicrobial interventions,which could contribute to sustainable farming practices.
基金supported by the National Natural Science Foundation of China(Nos.82171552 and 82170479)the Natural Science Foundation of Shanghai Ctiy(No.21ZR1457500)the Science and Technology Bureau of Shanghai Putuo District(No.ptkwws202102).
文摘Magnolol,a compound extracted from Magnolia officinalis,demonstrates potential efficacy in addressing metabolic dysfunction and cardiovascular diseases.Its biological activities encompass anti-inflammatory,antioxidant,anticoagulant,and anti-diabetic effects.Growth/differentiation factor-15(GDF-15),a member of the transforming growth factorβsuperfamily,is considered a potential therapeutic target for metabolic disorders.This study investigated the impact of magnolol on GDF-15 production and its underlying mechanism.The research examined the pharmacological effect of magnolol on GDF-15 expression in vitro and in vivo,and determined the involvement of endoplasmic reticulum(ER)stress signaling in this process.Luciferase reporter assays,chromatin immunoprecipitation,and in vitro DNA binding assays were employed to examine the regulation of GDF-15 by activating transcription factor 4(ATF4),CCAAT enhancer binding proteinγ(CEBPG),and CCCTC-binding factor(CTCF).The study also investigated the effect of magnolol and ATF4 on the activity of a putative enhancer located in the intron of the GDF-15 gene,as well as the influence of single nucleotide polymorphisms(SNPs)on magnolol and ATF4-induced transcription activity.Results demonstrated that magnolol triggers GDF-15 production in endothelial cells(ECs),hepatoma cell line G2(HepG2)and hepatoma cell line 3B(Hep3B)cell lines,and primary mouse hepatocytes.The cooperative binding of ATF4 and CEBPG upstream of the GDF-15 gene or the E1944285 enhancer located in the intron led to full-power transcription of the GDF-15 gene.SNP alleles were found to impact the magnolol and ATF4-induced transcription activity of GDF-15.In high-fat diet ApoE^(-/-)mice,administration of magnolol induced GDF-15 production and partially suppressed appetite through GDF-15.These findings suggest that magnolol regulates GDF-15 expression through priming of promoter and enhancer activity,indicating its potential as a drug for the treatment of metabolic disorders.
文摘BACKGROUND The Mac-2 binding protein glycosylated isomer(M2BPGi)is a serum marker for fibrosis that correlates with the fibrosis stages in various liver diseases.AIM To examine the M2BPGi’s threshold for staging fibrosis in patients with chronic hepatitis B(CHB),and its changes during treatment.METHODS This was a prospective,longitudinal study.A total of 348 eligible patients were recruited from the Hepatology Department,Medic Medical Center between March 2020 and December 2023.Liver enzyme tests,platelet counts,M2BPGi levels,and FibroScan were conducted at baseline and at 3-month intervals until six months post-treatment.Correlation plots of M2BPGi,FibroScan,and the other parameters were generated.Receiver operating characteristic curves were constructed for M2BPGi and the other parameters to evaluate their performance.RESULTS M2BPGi levels correlated well with FibroScan results and increased as the fibrosis stage advanced.The median M2BPGi levels at the different stages of fibrosis showed statistically significant differences.The cut-off values of M2BPGi for diagnosing significant fibrosis(F≥2),advanced fibrosis(F3),and cirrhosis(F4)were determined to be 1.08,1.4,and 1.52,respectively.In the context of fibrosis regression in CHB patients during the first 6-month of treatment,M2BPGi levels appeared to decrease before this pattern occurred in the FibroScan results.CONCLUSION M2BPGi levels were strongly correlated with FibroScan.M2BPGi can be used to assess liver fibrosis,and to serve as a tool for monitoring fibrosis regression in CHB patients undergoing treatment.
文摘BACKGROUND Cirrhosis is a progressive condition characterized by fibrosis that can lead to severe complications and increased mortality.The mac-2 binding protein glyco-sylation isomer(M2BPGi)is a prominent biomarker for predicting hepatocellular carcinoma(HCC)and cirrhosis-induced esophageal varices(EV).AIM To investigate thresholds of M2BPGi associated with HCC,EV,and decomp-ensation in patients with cirrhosis.METHODS This was a prospective study.A total of 153 patients with cirrhosis who met the inclusion criteria were enrolled.The patients were diagnosed with HCC and EV according to the Baveno VII and European Association for the Study of the Liver guidelines.Baseline serum M2BPGi levels were assessed along with other routine tests.The data analysis aimed to determine the cutoff values of M2BPGi for pre-dicting EV and HCC.RESULTS In the study 85.6%of patients were Child-Pugh B and C.M2BPGi mean cutoff index was 7.1±3.7,showing no significant etiological differences.However,M2BPGi levels varied significantly among Child-Pugh classes,EV classifications,and between patients with and without HCC(P<0.01).M2BPGi cutoff values for predicting HCC,EV,and decompensated cirrhosis were 6.50,6.64,and 5.25,respectively.Mul-tivariate analysis confirmed M2BPGi as an independent risk factor for EV[adjusted odds ratio(aOR):1.3,95%CI:1.08-1.64]and liver decompensation(aOR:2.11,95%CI:1.37-3.83).Area under the curve of M2BPGi for HCC differ-entiation was 0.71.An algorithm combining alpha-fetoprotein(AFP)and M2BPGi detected 26 of 28 HCC cases with 98.04%accuracy vs 10 cases by AFP alone.CONCLUSION Serum M2BPGi predicted cirrhosis complications,including decompensation and varices,especially in HCC.Combined with AFP,it enhanced HCC detection.Future liver biopsy studies are needed for tissue confirmation.
文摘BACKGROUND Mac-2 binding protein glycosylation isomer(M2BPGi)serves as a marker of activated hepatic stellate cells and as such holds potential as a biomarker for liver fibrosis.In Viet Nam,metabolic dysfunction-associated steatotic liver disease(MASLD)is rising in prevalence and there is an urgent need for better clinical management,particularly in early detection methods that will improve overall prognosis.AIM To examine M2BPGi cut-off values for staging liver fibrosis in patients with MASLD and risk factors associated with disease progression.METHODS A total of 301 individuals with ultrasound-confirmed or FibroScan-confirmed diagnosis of fatty liver were enrolled in the study.The participants were stratified according to fibrosis stage,measured via magnetic resonance elastography.M2-BPGi,Fibrosis-4(FIB-4)Index score,and routine parameters of liver function were assessed to statistically investigate the correlation of M2BPGi levels in various fibrosis stages and to identify risk factors associated with fibrosis severity.RESULTS M2BPGi levels positively correlated with fibrosis stages,with cut-off indexes of 0.57 for F0-1,0.68 for F2-3,and 0.78 for F4.M2BPGi levels in the F0-1 group were significantly different from those in both the F2-3 group(P=0.038)and the F4 group(P=0.0051);the F2-3 and F4 groups did not show a significant difference(P=0.39).Females exhibited significantly higher M2BPGi levels than males for all fibrosis stages,particularly in the F2-3 group(P=0.01)and F4 group(P=0.0006).In the F4(cirrhosis)group,individuals with diabetes had significantly higher M2BPGi levels than those without.M2BPGi,hemoglobin A1c,and FIB-4 score were identified as independent risk factors for greater fibrosis and cirrhosis.CONCLUSION M2BPGi levels varied significantly throughout fibrosis progression,from early MASLD to cirrhosis,with sex correlation.M2BPGi holds promise as an early biomarker for fibrosis characterization in MASLD adult patient populations.
基金supported by NSF 2333230 (J.L.),NIH National Institute of Dental and Craniofacial Research (NIDCR) awards,DE030943 (X.H.),DE023810 (X.H.) and DE031329 (J.L.),T90 DE026110,and K99 DE033794 (to P.-T.D.)
文摘Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome,with implications for microbial pathogenesis and host defense.Among these,transfer RNA-derived small RNAs(tsRNAs)have garnered attention for their roles in modulating microbial behavior.However,the bacterial factors mediating tsRNA interaction and functionality remain poorly understood.In this study,using RNA affinity pull-down assay in combination with mass spectrometry,we identified a putative membrane-bound protein,annotated as P-type ATPase transporter(PtaT)in Fusobacterium nucleatum(Fn),which binds Fn-targeting tsRNAs in a sequence-specific manner.Through targeted mutagenesis and phenotypic characterization,we showed that in both the Fn type strain and a clinical tumor isolate,deletion of ptaT led to reduced tsRNA intake and enhanced resistance to tsRNA-induced growth inhibition.Global RNA sequencing and label-free Raman spectroscopy revealed the phenotypic differences between Fn wild type and PtaT-deficient mutant,highlighting the functional significance of PtaT in purine and pyrimidine metabolism.Furthermore,AlphaFold 3 prediction provides evidence supporting the specific binding between PtaT and Fn-targeting tsRNA.By uncovering the first RNA-binding protein in Fn implicated in growth modulation through interactions with host-derived small RNAs(sRNAs),our study offers new insights into sRNA-mediated host-pathogen interplay within the context of microbiome-host interactions.
基金Supported by the Talent Launch Fund of Chongqing Medical University Affiliated University City HospitalChongqing Medical University Affiliated University City Hospital Youth Program,No.2021ZD05.
文摘BACKGROUND The exact mechanisms underlying diabetic nephropathy(DN)remain incompletely elucidated,prompting researchers to explore new perspectives and identify novel intervention targets in this field.AIM To explore the role and underlying mechanisms of farnesoid X receptor(FXR)in the development of DN by regulating endoplasmic reticulum stress(ERS)molecular chaperone binding immunoglobulin protein(BiP)expression.METHODS Bioinformatics analyses identified potential FXR-binding elements in the BiP promoter.Dual-luciferase and chromatin immunoprecipitation(ChIP)assays confirmed FXR-BiP binding sites.In vitro studies used SV40 MES 13 cells under varying glucose conditions and treatments with FXR modulators[obeticholic acid(INT-747)and guggulsterones]or BiP small interfering RNA.The expression of BiP and ERS-related proteins[protein kinase R-like endoplasmic reticulum kinase(PERK),inositol-requiring enzyme 1(IRE1),activating transcription factor 6(ATF6)]was assessed alongside cell proliferation and extracellular matrix(ECM)synthesis.In vivo studies in DN mice(db/db)examined the effects of FXR activation on renal function and morphology.RESULTS FXR bound to the target sequence in the BiP promoter region,enhancing transcriptional activity,as confirmed by ChIP experiments.FXR expression decreased in SV40 MES 13 cells stimulated with high glucose and in renal tissues of DN mice compared with control.Treatment of SV40 MES 13 cells with the FXR agonist INT-747 significantly increased intracellular BiP expression,whereas silencing the FXR gene led to the downregulation of BiP levels.In vivo administration of INT-747 significantly elevated BiP levels in renal tissues,improved renal function and fibrosis in DN mice,while inhibiting the expression of ERS-related signaling proteins PERK,IRE1,and ATF6.CONCLUSION FXR promotes BiP expression by binding to its promoter,suppressing ERS pathways,and reducing mesangial cell proliferation and ECM synthesis.These findings highlight FXR as a potential therapeutic target for diabetic glomerulosclerosis.
基金Supported by the National Research Foundation of Korea Grant Funded by the Korea Government,No.RS-2024-00440477the Korea Institute of Science and Technology Institutional Program,No.2E33111-24-042.
文摘BACKGROUND Mixed lineage kinase domain-like protein(MLKL)serves as a critical mediator in necroptosis,a form of regulated cell death linked to various liver diseases.This study aims to specifically investigate the role of MLKL’s adenosine triphosphate(ATP)-binding pocket in facilitating necroptosis-independent pathways that may contribute to liver disease progression.By focusing on this mechanism,we seek to identify potential therapeutic targets that can modulate MLKL activity,offering new strategies for the prevention and treatment of liver-related pathologies.AIM To investigate the possibility of using the ATP-binding pocket-associated,necro-ptosis-independent MLKL pathway as a target for liver diseases.METHODS Cell death following necroptosis stimuli was evaluated using cell proliferation assays,flow cytometry,and electron microscopy in various cells.The human liver organoid system was used to evaluate whether the MLKL ATP pocket-binding inhibitor could attenuate inflammation.Additionally,alcoholic and non-alcoholic fatty liver diseases animal models were used to determine whether MLKL ATP pocket inhibitors could attenuate liver injury.RESULTS While an MLKL ATP pocket-binding inhibitor did not prevent necroptosis-induced cell death in RAW 264.7 cells,it did reduce the necroptosis-led expression of CXCL2,ICAM,and VCAM.Notably,MLKL ATP pocket inhibitor diminishes the expression of CXCL2,ICAM,and VCAM by inhibiting the IκB kinase and nuclear factor kappa-B pathways without inducing necroptosis-induced cell death in two-dimensional cell culture as well as the human-derived liver organoid system.Although MLKL ATP-binding inhibitor was ineffective in non-alcoholic fatty liver disease animal models,MLKL ATP-binding inhibitor attenuated hepatic inflammation in the alcoholic liver disease model.CONCLUSION MLKL ATP pocket-binding inhibitor exerted anti-inflammatory effects through the necroptosis-independent MLKL pathway in an animal model of alcoholic liver disease.
文摘Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBIRC6)on non-small cell lung cancer(NSCLC)progression.Methods:The NSCLC and adjacent non-tumor tissues were collected at Shanghai Ninth People's Hospital.Quantitative real-time polymerase chain reaction was conducted for assessing the levels of circBIRC6,amyloid beta precursor protein binding protein 2(APPBP2)messenger RNA(mRNA),baculoviral IAP repeat containing 6 mRNA(BIRC6),and microRNA-217(miR-217).Western blot assay was adopted for measuring the protein levels of APPBP2,E-cadherin,N-cadherin,and vimentin.Colony formation assay,transwell assay,and flow cytometry analysis were utilized for evaluating cell colony formation,metastasis,and apoptosis.Dualluciferase reporter assay and RNA immunoprecipitation assay were carried out to determine the interaction between miR-217 and circBIRC6 and APPBP2 in NSCLC tissues.The murine xenograft model assay was used to investigate the function of circBIRC6 in tumor formation in vivo.Differences were analyzed via Student's t test or one-way analysis of variance.Pearson's correlation coefficient analysis was used to analyze linear correlation.Results:CircBIRC6 was overexpressed in NSCLC tissues and cells.Knockdown of circBIRC6 repressed the colony formation and metastasis and facilitated apoptosis of NSCLC cells in vitro and restrained tumorigenesis in vivo.Mechanically,circBIRC6 functioned as miR-217 sponge to promote APPBP2 expression in NSCLC cells.MiR-217 inhibition rescued circBIRC6 knockdown-mediated effects on NSCLC cell colony formation,metastasis,and apoptosis.Overexpression of miR-217 inhibited the malignant phenotypes of NSCLC cells,while the effects were abrogated by elevating APPBP2.Conclusion:CircBIRC6 aggravated NSCLC cell progression by elevating APPBP2 via sponging miR-217,which might provide a fresh perspective on NSCLC therapy.
基金Supported by Science Technology Research and Development Project in Shijiazhuang City in2010(10120803)Scientific Research Starting Fund Project of Shijiazhuang University in2007(2007012),Education Reform Research Item of Shijiazhuang University in2008(2008006)~~
文摘[Objective] The research aimed to find the extracellular binding proteins of CR4.[Method] The extracellular domain of OsCR4 was as the bait protein,and the yeast two-hybrid was used to screen cDNA library of seedling which was cultivated 14 d.[Result] A lot of proteins which included a peroxide B（D26484）,a methionine thioredoxin reductase（ABF96078）and an unknown function protein were gained.[Conclusion] It provided the theory basis for studying the signal transduction mechanism of CR4.
文摘The molecular mechanism of how hepatocytes maintain cholesterol homeostasis has become much more transparent with the discovery of sterol regulatory element binding proteins (SREBPs) in recent years. These membrane proteins aremembers of the basic helix-loop-helix-leucine zipper (bHLHZip) family of transcription factors. They activate the expression of at least 30 genes involved in the synthesis of cholesterol and lipids. SREBPs are synthesized as precursor proteins in the endoplasmic reticulum (ER), where they form a complex with another protein, SREBP cleavage activating protein (SCAP). The SCAP molecule contains a sterol sensory domain. In the presence of high cellular sterol concentrations SCAP confines SREBP to the ER. With low cellular concentrations, SCAP escorts SREBP to activation in the Golgi. There, SREBP undergoes two proteolytic cleavage steps to release the mature, biologically active transcription factor, nuclear SREBP (nSREBP). nSREBP translocates to the nucleus and binds to sterol response elements (SRE) in the promoter/enhancer regions of target genes. Additional transcription factors are required to activate transcription of these genes. Three different SREBPs are known, SREBPs-1a, -1c and -2. SREBP-1a and -1c are isoforms produced from a single gene by alternate splicing. SREBP-2 is encoded by a different gene and does not display any isoforms. It appears that SREBPs alone, in the sequence described above, can exert complete control over cholesterol synthesis, whereas many additional factors (hormones, cytokines, etc.) are required for complete control of lipid metabolism. Medicinal manipulation of the SREBP/SCAP system is expected to prove highly beneficial in the management of cholesterol-related disease.
文摘The association of retinol binding protein 4 （RBP4） with atherosclerosis of the carotid artery in type 2 diabetes mellitus （T2DM） remains undefined. We aimed to investigate the correlation of RBP4 expression with atherosclerosis of the carotid artery in T2DM. A total of 1,076 subjects were investigated for intima-media thickness of the bilateral common carotid arteries, and they were divided into three groups： in group Ⅰ, patients had normal neck vascular ultra- sound, in group Ⅱ, intimal carotid artery media thickness was equal to or more than 1 mm, and in group Ⅲ, carotid artery plaque was present. Height, weight, blood pressure （BP）, fasting plasma glucose （FPG）, hemoglobin Alc （HbA1c）, total cholesterol （TC）, triglyceride （TG）, low-density lipoprotein cholesterol （LDL-C）, high-density lipopro- tein cholesterol （HDL-C）, apolipoprotein A-1 （apoA-1）, apolipoprotein B （apoB） and lipoprotein （a） [Lp（a）] were determined by routine laboratory methods. RBP4 and high sensitivity C reactive protein （HsCRP） were measured by an enzyme-linked immuno-sorbent assay, and insulin concentration was measured by an electrochemiluminescence sandwich immunoassay. Duration of diabetes, waist and BP, FPG, HbAlc, TG, TC, LDL-C, APOB, Lp（a）, HsCRP, RBP4 and homeostasis model assessment insulin resistance index （HOMA-IR） were significantly lower in group I than in the other two groups （P〈0.01, P〈0.01）. Plasma levels of HbAlc, RBP4, LDL-C, TC, HOMA-IR, HsCRP and Lp（a）, waist and BP were significantly increased in group III than in group II （P〈0.01）. Multivariate logistic regression analysis showed that there were seven factors associated with the occurrence of carotid artery atherosclero- sis and its risks in descending order were： high LDL-C, high waist, high HsCRP, duration of diabetes, high HOMA-IR, HbAlc and high RBP4. Our finding supported that RBP4 was positively correlated with carotid atherosclerosis in patients with T2DM and could be used as an early predictor of cardiovascular disease.
基金supported by the National Natural Science Foundation of China(31271402 and 31100601)the National Key Basic Research Program(2012CB316503)
文摘Protein binding is essential to the transport,decay and regulation of almost all RNA molecules.However,the structural preference of protein binding on RNAs and their cellular functions and dynamics upon changing environmental conditions are poorly understood.Here,we integrated various high-throughput data and introduced a computational framework to describe the global interactions between RNA binding proteins(RBPs)and structured RNAs in yeast at single-nucleotide resolution.We found that on average,in terms of percent total lengths,~15%of mRNA untranslated regions(UTRs),~37%of canonical non-coding RNAs(ncRNAs)and^11%of long ncRNAs(lncRNAs)are bound by proteins.The RBP binding sites,in general,tend to occur at single-stranded loops,with evolutionarily conserved signatures,and often facilitate a specific RNA structure conformation in vivo.We found that four nucleotide modifications of tRNA are significantly associated with RBP binding.We also identified various structural motifs bound by RBPs in the UTRs of mRNAs,associated with localization,degradation and stress responses.Moreover,we identified>200 novel lncRNAs bound by RBPs,and about half of them contain conserved secondary structures.We present the first ensemble pattern of RBP binding sites in the structured non-coding regions of a eukaryotic genome,emphasizing their structural context and cellular functions.
基金supported by the National Basic Research Program of China(2012CB114104)the National Natural Science Foundation of China(30871640,31071694)+1 种基金the National High-Tech R&D Program of China(2008AA02Z307)the International Cooperation and Exchange Foundation of NSFC-RS of China(31111130203).
文摘The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,encoding a protein with 147 amino acids,namely HarmOBP5.HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography.SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics.Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles,including （E）-β-farnesene,ethyl butyrate,ethyl heptanoate,and acetic acid 2-methylbutyl ester.Based on three-dimensional （3D） model of AaegOBP1 from Aedes aegypti,a 3D model of HarmOBP5 was predicted.The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H.armigera.This study provides clues for better understanding physiological functions of OBPs in H.armigera and other insects.
