Trace amines(TAs)are metabolically related to catecholamine and associated with cancer and neurological disorders.Comprehensive measurement of TAs is essential for understanding pathological processes and providing pr...Trace amines(TAs)are metabolically related to catecholamine and associated with cancer and neurological disorders.Comprehensive measurement of TAs is essential for understanding pathological processes and providing proper drug intervention.However,the trace amounts and chemical instability of TAs challenge quantification.Here,diisopropyl phosphite coupled with chip two-dimensional(2D)liquid chromatography tandem triple-quadrupole mass spectrometry(LC-QQQ/MS)was developed to simultaneously determine TAs and associated metabolites.The results showed that the sensitivities of TAs increased up to 5520 times compared with those using nonderivatized LC-QQQ/MS.This sensitive method was utilized to investigate their alterations in hepatoma cells after treatment with sorafenib.The significantly altered TAs and associated metabolites suggested that phenylalanine and tyrosine metabolic pathways were related to sorafenib treatment in Hep3B cells.This sensitive method has great potential to elucidate the mechanism and diagnose diseases considering that an increasing number of physiological functions of TAs have been discovered in recent decades.展开更多
Sensitive detection and precise quantitation of trace-level crucial biomarkers in a complex sample matrix has become an important area of research.For example,the detection of high-sensitivity cardiac troponin I (hs-c...Sensitive detection and precise quantitation of trace-level crucial biomarkers in a complex sample matrix has become an important area of research.For example,the detection of high-sensitivity cardiac troponin I (hs-cTnI) is strongly recommended in clinical guidelines for early diagnosis of acute myocardial infarction.Based on the use of an electrode modified by single-walled carbon nanotubes (SWCNTs) and a Ru(bpy)32+-doped silica nanoparticle (Ru@SiO2)/tripropylamine (TPA) system,a novel type of electrochemiluminescent (ECL) magnetoimmunosensor is developed for ultrasensitive detection of hs-cTnI.In this approach,a large amount of[Ru(bpy)3]2+is loaded in SiO2(silica nanoparticles) as luminophores with high luminescent efficiency and SWCNTs as electrode surface modification material with excellent electrooxidation ability for TPA.Subsequently,a hierarchical micropillar array of microstructures is fabricated with a magnet placed at each end to efficiently confine a single layer of immunomagnetic microbeads on the surface of the electrode and enable 7.5-fold signal enhancement In particular,the use of transparent SWCNTs to modify a transparent ITO electrode provides a two-order-of-magnitude ECL signal amplification.A good linear calibration curve is developed for hs-cTnI concentrations over a wide range from 10 fg/ml to 10 ng/ml,with the limit of detection calculated as 8.720 fg/ml (S/N=3).This ultrasensitive immunosensor exhibits superior detection performance with remarkable stability,reproducibility,and selectivity.Satisfactory recoveries are obtained in the detection of hs-cTnI in human serum,providing a potentia analysis protocol for clinical applications.展开更多
热休克蛋白70羧基末端相互作用蛋白（carboxyl-terminus of the Hsp70 interacting protein，CHIP）是Ballinger等发现的同时具有辅助伴侣分子和E3泛素连接酶活性的特殊蛋白分子。作为辅助伴侣分子，它与Hsp90和Hsc／Hsp70相互作用，调...热休克蛋白70羧基末端相互作用蛋白（carboxyl-terminus of the Hsp70 interacting protein，CHIP）是Ballinger等发现的同时具有辅助伴侣分子和E3泛素连接酶活性的特殊蛋白分子。作为辅助伴侣分子，它与Hsp90和Hsc／Hsp70相互作用，调节Hsp介导的蛋白质的异常折叠；而作为E3泛素连接酶，通过泛素化-蛋白酶体途径参与许多疾病相关蛋白的降解过程。展开更多
Plants use a sophisticated immune system to perceive pathogen infection and activate immune responses in a tightly controlled manner.In barley,Hv WRKY2 acts as a repressor in barley disease resistance to the powdery m...Plants use a sophisticated immune system to perceive pathogen infection and activate immune responses in a tightly controlled manner.In barley,Hv WRKY2 acts as a repressor in barley disease resistance to the powdery mildew fungus,Blumeria graminis f.sp.hordei(Bgh).However,the molecular features of Hv WRKY2 in its DNA-binding and repressor functions,as well as its target genes,are uncharacterized.We show that the W-box binding of Hv WRKY2 requires an intact WRKY domain and an upstream sequence of~75 amino acids,and the Hv WRKY2 W-box binding activity is linked to its repressor function in disease resistance.Chromatin immunoprecipitation(ChIP)-seq analysis identified HvCEBiP,a putative chitin receptor gene,as a target gene of Hv WRKY2 in overexpressing transgenic barley plants.ChIP-qPCR and Electrophoretic Mobility Shift Assay(EMSA)verified the direct binding of Hv WRKY2 to a W-boxcontaining sequence in the HvCEBiP promoter.Hv CEBiP positively regulates resistance against Bgh in barley.Our findings suggest that Hv WRKY2 represses barley basal immunity by directly targeting pathogen-associated molecular pattern(PAMP)recognition receptor genes,suggesting that Hv CEBiP and likely chitin signaling function in barley PAMP-triggered immune responses to Bgh infection.展开更多
基金supported by the Science and Technology Development Fund,Macao,China(Grant No.:FDCT0044/2018/AFJ).
文摘Trace amines(TAs)are metabolically related to catecholamine and associated with cancer and neurological disorders.Comprehensive measurement of TAs is essential for understanding pathological processes and providing proper drug intervention.However,the trace amounts and chemical instability of TAs challenge quantification.Here,diisopropyl phosphite coupled with chip two-dimensional(2D)liquid chromatography tandem triple-quadrupole mass spectrometry(LC-QQQ/MS)was developed to simultaneously determine TAs and associated metabolites.The results showed that the sensitivities of TAs increased up to 5520 times compared with those using nonderivatized LC-QQQ/MS.This sensitive method was utilized to investigate their alterations in hepatoma cells after treatment with sorafenib.The significantly altered TAs and associated metabolites suggested that phenylalanine and tyrosine metabolic pathways were related to sorafenib treatment in Hep3B cells.This sensitive method has great potential to elucidate the mechanism and diagnose diseases considering that an increasing number of physiological functions of TAs have been discovered in recent decades.
基金The authors acknowledge financial support from the National Natural Science Foundation of China(Grant Nos.62001460,31971368,12202461,and 22104148)the Guangdong Regional Joint Funds for Young Scientists(Grant Nos.2020A1515110201 and 2020A1515110368)+2 种基金Guangdong Provincial General Funding(Grant No.2021A1515220156)Guangdong Basic and Applied Basic Research Funding-Regional Joint Fund(Grant No.2020B1515120040)Shenzhen Science and Technology Research Funding(Grant Nos.JSGG20201103153801005,JSGG20191115141601721,ZDSYS20220527171406014,JCYJ20220818101412027,JCYJ20200109115635440,and JCYJ 20200109115408041).
文摘Sensitive detection and precise quantitation of trace-level crucial biomarkers in a complex sample matrix has become an important area of research.For example,the detection of high-sensitivity cardiac troponin I (hs-cTnI) is strongly recommended in clinical guidelines for early diagnosis of acute myocardial infarction.Based on the use of an electrode modified by single-walled carbon nanotubes (SWCNTs) and a Ru(bpy)32+-doped silica nanoparticle (Ru@SiO2)/tripropylamine (TPA) system,a novel type of electrochemiluminescent (ECL) magnetoimmunosensor is developed for ultrasensitive detection of hs-cTnI.In this approach,a large amount of[Ru(bpy)3]2+is loaded in SiO2(silica nanoparticles) as luminophores with high luminescent efficiency and SWCNTs as electrode surface modification material with excellent electrooxidation ability for TPA.Subsequently,a hierarchical micropillar array of microstructures is fabricated with a magnet placed at each end to efficiently confine a single layer of immunomagnetic microbeads on the surface of the electrode and enable 7.5-fold signal enhancement In particular,the use of transparent SWCNTs to modify a transparent ITO electrode provides a two-order-of-magnitude ECL signal amplification.A good linear calibration curve is developed for hs-cTnI concentrations over a wide range from 10 fg/ml to 10 ng/ml,with the limit of detection calculated as 8.720 fg/ml (S/N=3).This ultrasensitive immunosensor exhibits superior detection performance with remarkable stability,reproducibility,and selectivity.Satisfactory recoveries are obtained in the detection of hs-cTnI in human serum,providing a potentia analysis protocol for clinical applications.
文摘热休克蛋白70羧基末端相互作用蛋白（carboxyl-terminus of the Hsp70 interacting protein，CHIP）是Ballinger等发现的同时具有辅助伴侣分子和E3泛素连接酶活性的特殊蛋白分子。作为辅助伴侣分子，它与Hsp90和Hsc／Hsp70相互作用，调节Hsp介导的蛋白质的异常折叠；而作为E3泛素连接酶，通过泛素化-蛋白酶体途径参与许多疾病相关蛋白的降解过程。
基金supported by National Key Research and Development Program of China(2018YFD1000703,2018YFD1000700)Strategic Priority Research Program of the Chinese Academy of Sciences(XDB11020400)+3 种基金National Program on Research and Development of Transgenic Plants(2016ZX08009-003-001)Startup Fund for Advanced Talents of Lushan Botanical GardenChinese Academy of Science(2020ZWZX03 and 2020ZWZX05)the“Double Hundred and Double Thousand”Talent Project of Jiujiang City(jjsbsq2020026)。
文摘Plants use a sophisticated immune system to perceive pathogen infection and activate immune responses in a tightly controlled manner.In barley,Hv WRKY2 acts as a repressor in barley disease resistance to the powdery mildew fungus,Blumeria graminis f.sp.hordei(Bgh).However,the molecular features of Hv WRKY2 in its DNA-binding and repressor functions,as well as its target genes,are uncharacterized.We show that the W-box binding of Hv WRKY2 requires an intact WRKY domain and an upstream sequence of~75 amino acids,and the Hv WRKY2 W-box binding activity is linked to its repressor function in disease resistance.Chromatin immunoprecipitation(ChIP)-seq analysis identified HvCEBiP,a putative chitin receptor gene,as a target gene of Hv WRKY2 in overexpressing transgenic barley plants.ChIP-qPCR and Electrophoretic Mobility Shift Assay(EMSA)verified the direct binding of Hv WRKY2 to a W-boxcontaining sequence in the HvCEBiP promoter.Hv CEBiP positively regulates resistance against Bgh in barley.Our findings suggest that Hv WRKY2 represses barley basal immunity by directly targeting pathogen-associated molecular pattern(PAMP)recognition receptor genes,suggesting that Hv CEBiP and likely chitin signaling function in barley PAMP-triggered immune responses to Bgh infection.
