Single-cell or low-input multi-omics techniques have revolutionized the study of pre-implantation embryo development.However,the single-cell or low-input proteomic research in this field is relatively underdeveloped b...Single-cell or low-input multi-omics techniques have revolutionized the study of pre-implantation embryo development.However,the single-cell or low-input proteomic research in this field is relatively underdeveloped because of the higher threshold of the starting material for mammalian embryo samples and the lack of hypersensitive proteome technology.In this study,a comprehensive solution of ultrasensitive proteome technology(CS-UPT)was developed for single-cell or low-input mouse oocyte/embryo samples.The deep coverage and high-throughput routes significantly reduced the starting material and were selected by investigators based on their demands.Using the deep coverage route,we provided the first large-scale snapshot of the very early stage of mouse maternal-to-zygotic transition,including almost 5,500 protein groups from 20 mouse oocytes or zygotes for each sample.Moreover,significant protein regulatory networks centered on transcription factors and kinases between the MII oocyte and 1-cell embryo provided rich insights into minor zygotic genome activation.展开更多
Elaioplasts of citrus peel are colorless plastids which accumulate significant amounts of terpenes.However,other functions of elaioplasts have not been fully characterized to date.Here,a LC–MS/MS shotgun technology w...Elaioplasts of citrus peel are colorless plastids which accumulate significant amounts of terpenes.However,other functions of elaioplasts have not been fully characterized to date.Here,a LC–MS/MS shotgun technology was applied to identify the proteins from elaioplasts that were highly purified from young fruit peel of kumquat.A total of 655 putative plastid proteins were identified from elaioplasts according to sequence homology in silico and manual curation.Based on functional classification via Mapman,~50%of the identified proteins fall into six categories,including protein metabolism,transport,and lipid metabolism.Of note,elaioplasts contained ATP synthase and ADP,ATP carrier proteins at high abundance,indicating important roles for ATP generation and transport in elaioplast biogenesis.Additionally,a comparison of proteins between citrus chromoplast and elaioplast proteomes suggest a high level of functional conservation.However,some distinctive protein profiles were also observed in both types of plastids notably for isoprene biosynthesis in elaioplasts,and carotenoid metabolism in chromoplasts.In conclusion,this comprehensive proteomic study provides new insights into the major metabolic pathways and unique characteristics of elaioplasts and chromoplasts in citrus fruit.展开更多
OBJECTIVE:To observe the efficacy of moxibustion in the treatment of chronic fatigue syndrome(CFS)and explore the effects on gut microbiota and metabolic profiles.METHODS:Forty-eight male Sprague-Dawley rats were rand...OBJECTIVE:To observe the efficacy of moxibustion in the treatment of chronic fatigue syndrome(CFS)and explore the effects on gut microbiota and metabolic profiles.METHODS:Forty-eight male Sprague-Dawley rats were randomly assigned to control group(Con),CFS model group(Mod,established by multiple chronic stress for 35 d),MoxA group(CFS model with moxibustion Shenque(CV8)and Guanyuan(CV4),10 min/d,28 d)and MoxB group(CFS model with moxibustion Zusanli(ST36),10 min/d,28 d).Open-field test(OFT)and Morris-water-maze test(MWMT)were determined for assessment the CFS model and the therapeutic effects of moxibustion.16S rRNA gene sequencing analysis based gut microbiota integrated untargeted liquid chromatograph-mass spectrometer(LC-MS)based fecal metabolomics were executed,as well as Spearman correlation analysis,was utilized to uncover the functional relevance between the potential metabolites and gut microbiota.RESULTS:The results of our behavioral tests showed that moxibustion improved the performance of CFS rats in the OFT and the MWMT.Microbiome profiling analysis revealed that the gut microbiomes of CFS rats were less diverse with altered composition,including increases in pro-inflammatory species(such as Proteobacteria)and decreases in anti-inflammatory species(such as Bacteroides,Lactobacillus,Ruminococcus,and Prevotella).Moxibustion partially normalized these changes in the gut microbiota.Furthermore,CFS was associated with metabolic disorders,which were effectively ameliorated by moxibustion.This was demonstrated by the normalization of 33 microbiota-related metabolites,including mannose(P=0.001),aspartic acid(P=0.009),alanine(P=0.007),serine(P=0.000),threonine(P=0.027),methionine(P=0.023),5-hydroxytryptamine(P=0.008),alpha-linolenic acid(P=0.003),eicosapentaenoic acid(P=0.006),hypoxanthine(P=0.000),vitamin B6(P=0.000),cholic acid(P=0.013),and taurocholate(P=0.002).Correlation analysis showed a significant association between the perturbed fecal microbiota and metabolite levels,with a notable negative relationship between LCA and Bacteroides.CONCLUSIONS:In this study,we demonstrated that moxibustion has an antifatigue-like effect.The results from the 16S rRNA gene sequencing and metabolomics analysis suggest that the therapeutic effects of moxibustion on CFS are related to the regulation of gut microorganisms and their metabolites.The increase in Bacteroides and decrease in LCA may be key targets for the moxibustion treatment of CFS.展开更多
The sub-cellular events that occur during the ethylene-modulated cell elongation were characterized by examining the ultra-structure of etiolated Arabidopsis seedling hypocotyl cells. Preventing the basal level ethyle...The sub-cellular events that occur during the ethylene-modulated cell elongation were characterized by examining the ultra-structure of etiolated Arabidopsis seedling hypocotyl cells. Preventing the basal level ethylene response facilitated cell elongation, and the cells exhibited wall loosening and separation phenotype. Nearby the wall separation sites were frequently associated with an increase in the cortical rough endoplasmic reticulum （rER） membranes, the presence of paramural bodies, and the circular Golgi formation. The cortical rER proliferation and circular Golgi phenotype were reverted by the protein biosynthesis inhibitor cycloheximide. The cortical rER membranes were longer when the ethylene response was prevented and shortened with elevated ethylene responses. Proteomic changes between wild type and the ethylene-insensitive mutant ethylene insensitive2 （ein2） seedling hypocotyls indicated that distinct subsets of proteins involving endomembrane trafficking, remodeling, and wall modifications were differentially expressed. FM4-64 staining supported the proteomic changes, which indicated reduced endocytosis activity with alleviation of the ethylene response. The basal level ethylene response has an important role in endomembrane trafficking, biological materials transport and maintenance of the endomembrane organization. It is possible that endomembrane alterations may partly associate with the wall modifications, though the biological significance of the alterations should be addressed in future studies.展开更多
基金supported by the National Natural Science Foundation of China(NSFC)(Grant Nos.:82030099,30700397 Detail)the National Key R&D Program of China(Grant No.:2022YFD2101500)+5 种基金the Science and Technology Commission of Shanghai Municipality,China(Grant No.:22DZ2303000)the Shanghai Municipal Science and Technology Commission“Science and Technology Innovation Action Plan”Technical Standard Project,China(Grant No.:21DZ2201700)the Shanghai Municipal Science and Technology Commission“Science and Technology Innovation Action Plan”Natural Science Foundation Project,China(Grant No.:23ZR1435800)the Strategic Priority Research Program of the Chinese Academy of Sciences,China(Grant No.:XDB32060000)the Basic Frontier Scientific Research Program of Chinese Academy of Sciences(Grant No.:ZDBS-LY-SM019)the Yangfan Project of Shanghai Science and Technology Commission,China(Grant No.:22YF1454100),and the Innovative Research Team of High-level Local Universities in Shanghai,China.
文摘Single-cell or low-input multi-omics techniques have revolutionized the study of pre-implantation embryo development.However,the single-cell or low-input proteomic research in this field is relatively underdeveloped because of the higher threshold of the starting material for mammalian embryo samples and the lack of hypersensitive proteome technology.In this study,a comprehensive solution of ultrasensitive proteome technology(CS-UPT)was developed for single-cell or low-input mouse oocyte/embryo samples.The deep coverage and high-throughput routes significantly reduced the starting material and were selected by investigators based on their demands.Using the deep coverage route,we provided the first large-scale snapshot of the very early stage of mouse maternal-to-zygotic transition,including almost 5,500 protein groups from 20 mouse oocytes or zygotes for each sample.Moreover,significant protein regulatory networks centered on transcription factors and kinases between the MII oocyte and 1-cell embryo provided rich insights into minor zygotic genome activation.
基金This work was supported by the National Natural Science Foundation of China(NSFC,grant nos.31501739)the National Basic Research Program of China(973 project No.2013CB127105)Huazhong Agricultural University Scientific&Technological Self-innovation Foundation(No.2662015BQ034).
文摘Elaioplasts of citrus peel are colorless plastids which accumulate significant amounts of terpenes.However,other functions of elaioplasts have not been fully characterized to date.Here,a LC–MS/MS shotgun technology was applied to identify the proteins from elaioplasts that were highly purified from young fruit peel of kumquat.A total of 655 putative plastid proteins were identified from elaioplasts according to sequence homology in silico and manual curation.Based on functional classification via Mapman,~50%of the identified proteins fall into six categories,including protein metabolism,transport,and lipid metabolism.Of note,elaioplasts contained ATP synthase and ADP,ATP carrier proteins at high abundance,indicating important roles for ATP generation and transport in elaioplast biogenesis.Additionally,a comparison of proteins between citrus chromoplast and elaioplast proteomes suggest a high level of functional conservation.However,some distinctive protein profiles were also observed in both types of plastids notably for isoprene biosynthesis in elaioplasts,and carotenoid metabolism in chromoplasts.In conclusion,this comprehensive proteomic study provides new insights into the major metabolic pathways and unique characteristics of elaioplasts and chromoplasts in citrus fruit.
基金National Natural Science Foundation of China:Based on TGF-β/Smad Signaling Pathway to Studying the Role and Mechanism of NF-κB in the Treatment of Chronic Fatigue Syndrome with Electroacupuncture (No.81704170)National Natural Science Foundation of China:Mechanism Study on Electroacupuncture Regulating MDM2 Ubiquitination PSD-95 Level and Remodeling Synaptic Structure of Hippocampal Neurons to Improve Cognitive Dysfunction in CFS (No.81873378)+7 种基金the Joint Guiding Project of Natural Science Foundation of Heilongjiang Province:Study on the Mechanism of Electroacupuncture Adjusting Gut Microbiota to Improve Oxidative Inflammatory Response in the Treatment of Chronic Fatigue Syndrome (No.LH2020H092)The Program of Heilongjiang Postdoctoral Initiation Fund:Experimental Study on Electroacupuncture Improving Protein Expression Differences and Target Effects in Hippocampus and Hypothalamus of Rats with Chronic Fatigue Syndrome (No.LBH-Q18117)Heilongjiang Administration of Traditional Chinese Medicine:Study on the Effect and Mechanism of Moxibustion on Gut Microbiota in Rats with Chronic Fatigue Syndrome (No.ZHY2020-79)The Mechanism of TGF-β/Smad Mediated NF-κB Inflammatory Pathway in the Treatment of Chronic Fatigue Syndrome by Electroacupuncture (No.ZHY16-003)The Mechanism of Electroacupuncture Improving Hippocampal Synaptic Plasticity in CFS Cognitive Dysfunction Rats (ZHY2022-136)Zhejiang Chinese Medical University Scientific Research:Study on the Mechanism of Moxibustion Regulating the Gut Microbiota of Chronic Fatigue Syndrome Targeting the 5-HT Signal System (No.2021RCZXZK02)Basic Public Welfare Project of the Third Affiliated Hospital of Zhejiang Chinese Medical University:Study on the Mechanism of Moxibustion Repairing the Barrier Function-Inflammatory Response of Chronic Fatigue Syndrome by the Gut-Brain Axis (No.ZS21ZA08)Scientific Research Foundation of Heilongjiang University of Chinese Medicine:Based on Gut Microbiota and Metabolism to Study the Mechanism in the Treatment of Chronic Fatigue Syndrome by Moxibustion (No.2019BS03)
文摘OBJECTIVE:To observe the efficacy of moxibustion in the treatment of chronic fatigue syndrome(CFS)and explore the effects on gut microbiota and metabolic profiles.METHODS:Forty-eight male Sprague-Dawley rats were randomly assigned to control group(Con),CFS model group(Mod,established by multiple chronic stress for 35 d),MoxA group(CFS model with moxibustion Shenque(CV8)and Guanyuan(CV4),10 min/d,28 d)and MoxB group(CFS model with moxibustion Zusanli(ST36),10 min/d,28 d).Open-field test(OFT)and Morris-water-maze test(MWMT)were determined for assessment the CFS model and the therapeutic effects of moxibustion.16S rRNA gene sequencing analysis based gut microbiota integrated untargeted liquid chromatograph-mass spectrometer(LC-MS)based fecal metabolomics were executed,as well as Spearman correlation analysis,was utilized to uncover the functional relevance between the potential metabolites and gut microbiota.RESULTS:The results of our behavioral tests showed that moxibustion improved the performance of CFS rats in the OFT and the MWMT.Microbiome profiling analysis revealed that the gut microbiomes of CFS rats were less diverse with altered composition,including increases in pro-inflammatory species(such as Proteobacteria)and decreases in anti-inflammatory species(such as Bacteroides,Lactobacillus,Ruminococcus,and Prevotella).Moxibustion partially normalized these changes in the gut microbiota.Furthermore,CFS was associated with metabolic disorders,which were effectively ameliorated by moxibustion.This was demonstrated by the normalization of 33 microbiota-related metabolites,including mannose(P=0.001),aspartic acid(P=0.009),alanine(P=0.007),serine(P=0.000),threonine(P=0.027),methionine(P=0.023),5-hydroxytryptamine(P=0.008),alpha-linolenic acid(P=0.003),eicosapentaenoic acid(P=0.006),hypoxanthine(P=0.000),vitamin B6(P=0.000),cholic acid(P=0.013),and taurocholate(P=0.002).Correlation analysis showed a significant association between the perturbed fecal microbiota and metabolite levels,with a notable negative relationship between LCA and Bacteroides.CONCLUSIONS:In this study,we demonstrated that moxibustion has an antifatigue-like effect.The results from the 16S rRNA gene sequencing and metabolomics analysis suggest that the therapeutic effects of moxibustion on CFS are related to the regulation of gut microorganisms and their metabolites.The increase in Bacteroides and decrease in LCA may be key targets for the moxibustion treatment of CFS.
基金supported by the National Natural Sciences Foundation of China (31070249, 31100212, and 31123006)the Chinese Ministry of Science and Technology (2011CB100700 and 2012AA10A302-2)the Chinese Academy of Sciences (KSCX2-EW-J-12)
文摘The sub-cellular events that occur during the ethylene-modulated cell elongation were characterized by examining the ultra-structure of etiolated Arabidopsis seedling hypocotyl cells. Preventing the basal level ethylene response facilitated cell elongation, and the cells exhibited wall loosening and separation phenotype. Nearby the wall separation sites were frequently associated with an increase in the cortical rough endoplasmic reticulum （rER） membranes, the presence of paramural bodies, and the circular Golgi formation. The cortical rER proliferation and circular Golgi phenotype were reverted by the protein biosynthesis inhibitor cycloheximide. The cortical rER membranes were longer when the ethylene response was prevented and shortened with elevated ethylene responses. Proteomic changes between wild type and the ethylene-insensitive mutant ethylene insensitive2 （ein2） seedling hypocotyls indicated that distinct subsets of proteins involving endomembrane trafficking, remodeling, and wall modifications were differentially expressed. FM4-64 staining supported the proteomic changes, which indicated reduced endocytosis activity with alleviation of the ethylene response. The basal level ethylene response has an important role in endomembrane trafficking, biological materials transport and maintenance of the endomembrane organization. It is possible that endomembrane alterations may partly associate with the wall modifications, though the biological significance of the alterations should be addressed in future studies.
