Perfluorooctanoic acid(PFOA)is a novel type of persistent synthetic organic pollutant,and its exposure on pregnant women can cause some adverse effects,such as pregnancy-induced hypertension,gestational diabetes melli...Perfluorooctanoic acid(PFOA)is a novel type of persistent synthetic organic pollutant,and its exposure on pregnant women can cause some adverse effects,such as pregnancy-induced hypertension,gestational diabetes mellitus,and preeclampsia.Therefore,understanding the metabolic changes caused by PFOA exposure is of great significance to protect pregnant women from its adverse effects.In this study,the metabolomes from the urine samples of pregnant women exposure to PFOA at different degrees were analyzed by GC-MS and LC-MS.The samples in different groups were distinguished and the differential metabolites were screened based on the VIP value,FC,and P-value of each comparison group through multivariate statistical analysis.The pathways related to differential metabolites were searched to reveal the effects of PFOA exposure on metabolic changes in pregnant women at different degrees.Finally,the ROC of differential metabolites was performed,and the differential metabolites with large area under the curve(AUC)values were selected and compared to identify the mutually differential metabolites.Meanwhile,these metabolites were fitted with a multivariable to explore if they could be used to distinguish different groups.The quantitative comparison of mutually differential metabolites revealed that the levels of L-cysteine,glycine,and 5-aminovaleric acid were positively correlated with the degree of PFOA exposure,indicating that different degrees of PFOA exposure could affect the synthesis or degradation of GSH and change the metabolism of oral or intestinal microbiota.Additionally,they may cause oxidative stress and abnormal fat metabolism in pregnant women.展开更多
Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In...Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In the present study,a novel separation strategy for exosomes is proposed based on the specific interaction between immobilized peptide ligands and phosphatidylserine moieties which are highly abundant on the surface of exosomes.With the new affinity method,intact model exosomes can be recovered with a high yield in a short processing time.The purity of exosome samples enriched from serum by the affinity method is far higher than that isolated by ultrafiltration,and similar to that obtained by density gradient centrifugation and ultracentrifugation.Moreover,the variety of contaminants co-isolated by the affinity method is relatively low due to its specific separation principle.Proteomics analysis of exosomes isolated by the affinity method from the serum of healthy,hepatocellular carcinoma patients,and intrahepatic cholangiocarcinoma patients was performed to prove the applicability of this method.In conclusion,our novel strategy shows characteristics of easy preparation,high specificity,and cost-effectiveness,and provides a promising approach for exosome isolation which should have wide applications.展开更多
基金the National Natural Science Foundation of China(Nos.81874307 and 21874088)the Shanghai Science and Technology Commission Scientific Research Project(Nos.19142203100,20142200400,18490731500,and 19DZ2202200)the National Facility for Translational Medicine,Shanghai(No.TMSK-2020-131).
文摘Perfluorooctanoic acid(PFOA)is a novel type of persistent synthetic organic pollutant,and its exposure on pregnant women can cause some adverse effects,such as pregnancy-induced hypertension,gestational diabetes mellitus,and preeclampsia.Therefore,understanding the metabolic changes caused by PFOA exposure is of great significance to protect pregnant women from its adverse effects.In this study,the metabolomes from the urine samples of pregnant women exposure to PFOA at different degrees were analyzed by GC-MS and LC-MS.The samples in different groups were distinguished and the differential metabolites were screened based on the VIP value,FC,and P-value of each comparison group through multivariate statistical analysis.The pathways related to differential metabolites were searched to reveal the effects of PFOA exposure on metabolic changes in pregnant women at different degrees.Finally,the ROC of differential metabolites was performed,and the differential metabolites with large area under the curve(AUC)values were selected and compared to identify the mutually differential metabolites.Meanwhile,these metabolites were fitted with a multivariable to explore if they could be used to distinguish different groups.The quantitative comparison of mutually differential metabolites revealed that the levels of L-cysteine,glycine,and 5-aminovaleric acid were positively correlated with the degree of PFOA exposure,indicating that different degrees of PFOA exposure could affect the synthesis or degradation of GSH and change the metabolism of oral or intestinal microbiota.Additionally,they may cause oxidative stress and abnormal fat metabolism in pregnant women.
基金supported by the National Natural Science Foundation of China(81874307,21874088)Shanghai Science and Technology Commission Scientific Research Project(18142200700,19142203100,20142200400)+1 种基金the Open Project Program of Engineering Research Center of Cell&Therapeutic Antibody,Ministry of Education,Shanghai Jiao Tong University(19X110020009-005)Startup Fund for Youngman Research at SJTU(19X100040029).
文摘Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In the present study,a novel separation strategy for exosomes is proposed based on the specific interaction between immobilized peptide ligands and phosphatidylserine moieties which are highly abundant on the surface of exosomes.With the new affinity method,intact model exosomes can be recovered with a high yield in a short processing time.The purity of exosome samples enriched from serum by the affinity method is far higher than that isolated by ultrafiltration,and similar to that obtained by density gradient centrifugation and ultracentrifugation.Moreover,the variety of contaminants co-isolated by the affinity method is relatively low due to its specific separation principle.Proteomics analysis of exosomes isolated by the affinity method from the serum of healthy,hepatocellular carcinoma patients,and intrahepatic cholangiocarcinoma patients was performed to prove the applicability of this method.In conclusion,our novel strategy shows characteristics of easy preparation,high specificity,and cost-effectiveness,and provides a promising approach for exosome isolation which should have wide applications.