The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook ...The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents.To address this limitation,a fishing strategy based on offline two-dimensional liquid chromatography(2D-LC)combined with surface plasmon resonance(SPR)was utilized to screen bioactive trace components targeting peroxiredoxin 3(PRDX3),using Uncaria alkaloids(UAs)as a case study.Initially,an orthogonal preparative offline 2D-LC system combining a positively charged C_(18)column and a conventional C_(18)column under disparate mobile phase conditions was constructed.To fully reveal the trace alkaloids,132D fractions of UAs were prepared,and their components were characterized using mass spectrometry(MS).Subsequently,employing PRDX3 as the targeting protein,a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether(GSM)serving as a positive control for binding.Employing this refined strategy,29 candidate binding alkaloids were fished from the 132D fractions.Notably,combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone.Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids,with isovallesiachotamine(IV),corynoxeine N-oxide(CO-N),and cadambine(CAD)demonstrating the highest affinity for PRDX3.Their interactions were further validated through molecular docking analysis.Subsequent intracellular H_(2)O_(2)measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H_(2)O_(2)clearance.In conclusion,this study introduced an innovative strategy for the identification of active trace components in TCM.This approach holds promise for accelerating research on medicinal components within this field.展开更多
Trace metals in PM2.5 were measured at one industrial site and one urban site during September, 2010 in Ji＇nan, eastern China. Individual aerosol particles and PM2.5 samples were collected concurrently at both sites....Trace metals in PM2.5 were measured at one industrial site and one urban site during September, 2010 in Ji＇nan, eastern China. Individual aerosol particles and PM2.5 samples were collected concurrently at both sites. Mass concentrations of eleven trace metals （i.e., Al, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Sr, Ba, and Pb） and one metalloid （i.e., As） were measured by inductively coupled plasma atomic emission spectroscopy （ICP-AES）. The result shows that mass concentrations of PM2.5 （130μg/m3） and trace metals （4.03 μg/m3） at the industrial site were 1.3 times and 1.7 times higher than those at the urban site, respectively, indicating that industrial activities nearby the city can emit trace metals into the surrounding atmosphere. Fe concentrations were the highest among all the measured trace metals at both sites, with concentrations of 1.04 ixg/m 3 at the urban site and 2.41 Itg/m3 at the industrial site, respectively. In addition, Pb showed the highest enrichment factors at both sites, suggesting the emissions from anthropogenic activities existed around the city. Correlation coefficient analysis and principal component analysis revealed that Cu, Fe, Mn, Pb, and Zn were originated from vehicular traffic and industrial emissions at both sites; As, Cr, and part of Pb from coal-fired power plant; Ba and Ti from natural soil. Based on the transmission electron microscopy analysis, we found that most of the trace metals were internally mixed with secondary sulfate/organic particles. These internally mixed trace metals in the urban air may have different toxic abilities compared with externally mixed trace metals.展开更多
Trace metals associated with PM10 aerosols and their variation during day and nighttime as well as during different seasons have been studied for the year 2012. PCA analysis suggested5 PCs,which accounted for 86.8% cu...Trace metals associated with PM10 aerosols and their variation during day and nighttime as well as during different seasons have been studied for the year 2012. PCA analysis suggested5 PCs,which accounted for 86.8% cumulative variance. PC1 accounted for 30% with a significant loading of metals of anthropogenic origin,while PC2 showed 28% variance with the loading of metals of crustal origin. These trace metals showed seasonal distinct day and night time characteristics. The concentrations of Cu,Pb,and Cd were found to be higher during nighttime in all the seasons. Only Fe was observed with significantly higher mean concentrations during daytime of all seasons except monsoon. The highest mean values of Cu,Cd,Zn,and Pb during post-monsoon might be attributed to winds advection over the regions of waste/biomass burning and industrial activities in Punjab and Haryana regions.Furthermore,concentration weighted trajectory analysis suggested that metals of crustal origin were contributed by long-range transport while metals of anthropogenic and industrial activities were contributed by regional/local source regions.展开更多
基金supported by Shanghai Institute of Materia Medica-Shanghai University of Traditional Chinese Medicine(SIMM-SHUTCM)Traditional Chinese Medicine Innovation Joint Research Program,China(Grant No.:2022,E2G808H096)the National Key Research and Development Program of China(Grant Nos.:2022YFC3501704 and 2023YFC3504205)Sanming Project of Medicine in Shenzhen,China(Grant No.:ZZYSM202106004).
文摘The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents.To address this limitation,a fishing strategy based on offline two-dimensional liquid chromatography(2D-LC)combined with surface plasmon resonance(SPR)was utilized to screen bioactive trace components targeting peroxiredoxin 3(PRDX3),using Uncaria alkaloids(UAs)as a case study.Initially,an orthogonal preparative offline 2D-LC system combining a positively charged C_(18)column and a conventional C_(18)column under disparate mobile phase conditions was constructed.To fully reveal the trace alkaloids,132D fractions of UAs were prepared,and their components were characterized using mass spectrometry(MS).Subsequently,employing PRDX3 as the targeting protein,a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether(GSM)serving as a positive control for binding.Employing this refined strategy,29 candidate binding alkaloids were fished from the 132D fractions.Notably,combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone.Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids,with isovallesiachotamine(IV),corynoxeine N-oxide(CO-N),and cadambine(CAD)demonstrating the highest affinity for PRDX3.Their interactions were further validated through molecular docking analysis.Subsequent intracellular H_(2)O_(2)measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H_(2)O_(2)clearance.In conclusion,this study introduced an innovative strategy for the identification of active trace components in TCM.This approach holds promise for accelerating research on medicinal components within this field.
基金supported by the National Basic Research Program(973)of China(No.2011CB403401)the National Natural Science Foundation of China(No.41105088,41275141)+1 种基金the Natural Science Foundation of Shandong Province(No.ZR2011DQ001)the State Key Laboratory for Coal Resources and Safe Mining(No.SKLCRSM11KFB03)
文摘Trace metals in PM2.5 were measured at one industrial site and one urban site during September, 2010 in Ji＇nan, eastern China. Individual aerosol particles and PM2.5 samples were collected concurrently at both sites. Mass concentrations of eleven trace metals （i.e., Al, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Sr, Ba, and Pb） and one metalloid （i.e., As） were measured by inductively coupled plasma atomic emission spectroscopy （ICP-AES）. The result shows that mass concentrations of PM2.5 （130μg/m3） and trace metals （4.03 μg/m3） at the industrial site were 1.3 times and 1.7 times higher than those at the urban site, respectively, indicating that industrial activities nearby the city can emit trace metals into the surrounding atmosphere. Fe concentrations were the highest among all the measured trace metals at both sites, with concentrations of 1.04 ixg/m 3 at the urban site and 2.41 Itg/m3 at the industrial site, respectively. In addition, Pb showed the highest enrichment factors at both sites, suggesting the emissions from anthropogenic activities existed around the city. Correlation coefficient analysis and principal component analysis revealed that Cu, Fe, Mn, Pb, and Zn were originated from vehicular traffic and industrial emissions at both sites; As, Cr, and part of Pb from coal-fired power plant; Ba and Ti from natural soil. Based on the transmission electron microscopy analysis, we found that most of the trace metals were internally mixed with secondary sulfate/organic particles. These internally mixed trace metals in the urban air may have different toxic abilities compared with externally mixed trace metals.
基金Partial financial support from the CSIR network project(PSC 0112)CSIR fellowship to Subhash Chandra,SRF
文摘Trace metals associated with PM10 aerosols and their variation during day and nighttime as well as during different seasons have been studied for the year 2012. PCA analysis suggested5 PCs,which accounted for 86.8% cumulative variance. PC1 accounted for 30% with a significant loading of metals of anthropogenic origin,while PC2 showed 28% variance with the loading of metals of crustal origin. These trace metals showed seasonal distinct day and night time characteristics. The concentrations of Cu,Pb,and Cd were found to be higher during nighttime in all the seasons. Only Fe was observed with significantly higher mean concentrations during daytime of all seasons except monsoon. The highest mean values of Cu,Cd,Zn,and Pb during post-monsoon might be attributed to winds advection over the regions of waste/biomass burning and industrial activities in Punjab and Haryana regions.Furthermore,concentration weighted trajectory analysis suggested that metals of crustal origin were contributed by long-range transport while metals of anthropogenic and industrial activities were contributed by regional/local source regions.
