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GreenPhos,a universal method for in-depth measurement of plant phosphoproteomes with high quantitative reproducibility

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Molecular Plant 2024年第1期17卷 199-213页

作者： xiaoxiao duan Yuanya Zhang Xiahe Huang Xiao Ma Hui Gao Yan Wang Zhen Xiao Chengcheng Huang Zhongshu Wang Bolong Li Wenqiang Yang Yingchun Wang State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesBejing 100101China University of Chinese Academy of Sciences Bejing 100049China Key Laboratory of Photobiology Instute of BotanyChinese Academy of SciencesBejing 100093China

Protein phosphorylation regulates a variety of important cellular and physiological processes in plants.In-depth profiling of plant phosphoproteomes has been more technically challenging than that of animal phosphoproteomes.This is largely due to the need to improve protein extraction efficiency from plant cells,which have a dense cell wall,and to minimize sample loss resulting from the stringent sample clean-up steps required for the removal of a large amount of biomolecules interfering with phosphopeptide purification and mass spectrometry analysis.To this end,we developed a method with a streamlined workflow for highly efficient purification of phosphopeptides from tissues of various green organisms including Arabidopsis,rice,tomato,and Chlamydomonas reinhardtii,enabling in-depth identification with high quantitative reproducibility of about 11000 phosphosites,the greatest depth achieved so far with single liquid chromatography-mass spectrometry(LC-MS)runs operated in a data-dependent acquisition(DDA)mode.The mainstay features of the method are the minimal sample loss achieved through elimination of sample clean-up before protease digestion and of desalting before phosphopeptide enrichment and hence the dramatic increases of time-and cost-effectiveness.The method,named GreenPhos,combined with single-shot LC-MS,enabled in-depth quantitative identification of Arabidopsis phosphoproteins,including differentially phosphorylated spliceosomal proteins,at multiple time points during salt stress and a number of kinase substrate motifs.GreenPhos is expected to serve as a universal method for purification of plant phosphopeptides,which,if samples are further fractionated and analyzed by multiple LC-MS runs,could enable measurement of plant phosphoproteomes with an unprecedented depth using a given mass spectrometry technology.

关键词： plant phosphoproteomics phosphopeptides LC–MS Arabidopsis salt stress

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Substitutions of stem-loop subdomains in internal ribosome entry site of Senecavirus A:Impacts on rescue of sequence-modifying viruses

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Journal of Integrative Agriculture 2024年第7期23卷 2391-2406页

作者： Qianqian Wang Jie Wang Lei Zhang xiaoxiao duan Lijie Zhu Youming Zhang Yan Li Fuxiao Liu College of Veterinary Medicine Qingdao Agricultural UniversityQingdao 266109China College of Veterinary Medicine Inner Mongolia Agricultural UniversityHuhhot 010018China Shandong New Hope Liuhe Group Co. Ltd.Qingdao 266100China Qingdao Center for Animal Disease Control&Prevention Qingdao 266199China State Key Laboratory of Microbial Technology Shandong UniversityQingdao 266237China

Senecavirus A(SVA)has a positive-sense,single-stranded RNA genome.Its 5´untranslated region harbors an internal ribosome entry site(IRES),comprising 10 larger or smaller stem-loop structures(including a pseudoknot)that have been demonstrated to be well conserved.However,it is still unclear whether each stem-loop subdomain,such as a single stem or loop,is also highly conserved.To clarify this issue in the present study,a set of 29 SVA cDNA clones were constructed by site-directed mutagenesis(SDM)on the IRES.The SDM-modified scenarios included:(1)stem-formed complementary sequences exchanging with each other;(2)loop transversion;(3)loop transition;and(4)point mutations.All cDNA clones were separately transfected into cells for rescuing viable viruses,whereas only four SVAs of interest could be recovered,and were genetically stable during 20 passages.One progeny grew significantly slower than the other three did.The dual-luciferase reporter assay showed that none of the SDM-modified IRESes significantly inhibited the IRES activity.Our previous study indicated that a single motif from any of the ten stem structures,if completely mutated,would cause the failure of virus recovery.Interestingly,our present study revealed three stem structures,whose individual complementary sequences could exchange with each other to rescue sequence-modifying SVAs.Moreover,one apical loop was demonstrated to have the ability to tolerate its own full-length transition,also having no impact on the recovery of sequence-modifying SVA.The present study suggested that not every stem-loop structure was strictly conserved in its conformation,while the full-length IRES itself was well conserved.This provides a new research direction on interaction between the IRES and many factors.

关键词： SVA HCV IRES HCV-like IRES stem-loop structure cDNA clone virus rescue mutation

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A novel structure of ultra-high-loading small moleculesencapsulated ZIF-8 colloid particles

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Nano Research 2024年第4期17卷 2929-2940页

作者： Pengfei duan Yunhe An xiaoxiao Wei Yanjie Tian Di Guan Xiangwen Liu Lanqun Mao Institute of Analysis and Testing Beijing Academy of Science and Technology(Beijing Center for Physical and Chemical Analysis)Beijing 100089China College of Chemistry Beijing Normal UniversityBeijing 100875China

The safety of nanoparticle-based drug delivery systems(DDSs)for cancer treatment is still a challenge,restricted by the intrinsic cytotoxicity of drug carriers and leakage of loaded drug.Here,we propose a novel nanocarrier’s cytotoxicity avoidance strategy by synthesizing an encapsulation core–shell structure of zeolitic imidazolate framework-8(ZIF-8)-based colloid particles(CPs)with an amorphous ZIF-8 skin.This encapsulation structure achieves an ultra-high loading rate(LR)of 90%(i.e.,9 mg doxorubicin(DOX)per 1 mg ZIF-8)for DOX and the protection of DOX from leaking.Notably,to deliver unit-dose drug,this ultra-high LR of 90%significantly reduces the usage of ZIF-8 to 1.2%(2 orders of magnitude)compared to that of DOX@ZIF-8 with a 10%LR,in which cytotoxicity of ZIF-8 could well below the safety limit and then be relatively ignored.Safety,drug delivery efficacy,scale-up ability,and universality of this encapsulation structure have been further verified.Our findings suggest the great potential of this ZIF-8-based encapsulation core–shell structure in the field of drug delivery.

关键词： metal-organic frameworks zeolitic imidazolate framework-8(ZIF-8) drug delivery nanoparticles nanostructures

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Apigenin ameliorates imiquimod-induced psoriasis in C57BL/6J mice by inactivating STAT3 and NF-κB

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Food Science and Human Wellness 2024年第1期13卷 211-224页

作者： Xianshe Meng Shihong Zheng Zequn Yin Xuerui Wang Daigang Yang Tingfeng Zou Huaxin Li Yuanli Chen Chenzhong Liao Zhouling Xie Xiaodong Fan Jihong Han Yajun duan xiaoxiao Yang Key Laboratory of Metabolism and Regulation for Major Diseases of Anhui Higher Education Institutes College of Food and Biological EngineeringHefei University of TechnologyHefei 230601China Tianjin Key Laboratory of human Development and Reproductive Regulation Department of General GynecologyTianjin Central Hospital of Gynecology and ObstetricsTianjin 300100China Key Laboratory of Medicinal Chemical Biology Key Laboratory of Bioactive Materials of Ministry of EducationCollege of Life SciencesNankai UniversityTianjin 300071China

Psoriasis is a chronic autoimmune disease featured by patches on the skin.It is caused by malfunction of immune cells and keratinocytes with inflammation as one of its key features.Apigenin(API)is a natural flavonoid with anti-inflammatory and immunoregulatory properties.Therefore,we speculated that API can ameliorate psoriasis,and determined its effect on the development of psoriasis by using imiquimod(IMQ)-induced psoriasis mouse model.Our results showed that API attenuated IMQ-induced phenotypic changes,such as erythema,scaling and epidermal thickening,and improved splenic hyperplasia.Abnormal differentiation of immune cells was restored in API-treated mice.Mechanistically,we revealed that API is a key regulator of signal transducer activator of transcription 3(STAT3).API regulated immune responses by reducing interleukin-23(IL-23)/STAT3/IL-17A axis.Moreover,it suppressed IMQ-caused cell hyperproliferation by inactivating STAT3 through regulation of extracellular signal-regulated kinase 1/2 and nuclear factor-κB(NF-κB)pathway.Furthermore,API reduced expression of inflammatory cytokines through inactivation of NF-κB.Taken together,our study demonstrates that API can ameliorate psoriasis and may be considered as a strategy for psoriasis treatment.

关键词： Psoriasis Apigenin Imiquimod Inflammation Signal transducer activator of transcription 3 (STAT3) Nuclear factor-κB(NF-κB)

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Promotional effect of Cu additive for the selective catalytic oxidation of n-butylamine over CeZrO_(x)catalyst

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Chinese Chemical Letters 2022年第6期33卷 3065-3072页

作者： Xin Xing Ting Zhao Jie Cheng xiaoxiao duan Wenpeng Li Ganggang Li Zhongshen Zhang Zhengping Hao National Engineering Laboratory for VOCs Pollution Control Material&Technology Research Center for Environmental Material and Pollution Control TechnologyUniversity of Chinese Academy of SciencesBeijing 101408China College of Environmental Science and Engineering Taiyuan University of Science and TechnologyTaiyuan 030024China

The catalytic elimination of nitrogen-containing volatile organic compounds(NVOCs)still encounters bottlenecks in NO_(x)formation and low N_(2)selectivity.Here,a series of Cu-promoted Ce-Zr mixed oxide catalysts were synthesized using a simple precipitation approach,and n-butylamine was adopted as the probe pollutant to evaluate their catalytic performance.The Ce Cu_(10%)ZrO_(x)catalyst exhibited the best catalytic activity,with 100%n-butylamine conversion and 90%N_(2)selectivity at 250℃.Concurrently,this sample also displayed good water resistance.A detailed characterization of the catalyst was performed through a series of experimental studies and theoretical calculations.The addition of Cu increased the redox property and promoted the production of oxygen vacancies,all of which were favorable for the greatest nbutylamine selective catalytic oxidation performance.The changes of oxygen vacancies over Ce Cu_(10%)ZrO_(x)in reaction process were studied by in situ Raman spectra.Moreover,in situ diffuse reflectance infrared Fourier transform spectra(DRIFTs)and theoretical calculations were employed to explore the reaction mechanism of n-butylamine selective oxidation.The high activity and selectivity of this catalyst confirm the practical feasibility of the selective oxidation of n-butylamine to CO_(2)and N_(2),and the exploration of the reaction mechanism provides new insights into the further design of catalysts.

关键词： N-Butylamine CeCu_(a)ZrO_(x)mixed oxide Selective oxidation Oxygen vacancies Reaction mechanism

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L-Ergothioneine ameliorates MPTP-induced Parkinson's disease in C57BL/6J mice by activating DJ-1

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Food Science and Human Wellness 2024年

作者： Xianshe Meng# Huawen Meng# Shuzeng Hou Zequn Yin Xuerui Wang Ke Gong Feng Zhang Qingshan Li Shuang Zhang Yuanli Chen xiaoxiao Yang Zhiwei Zhao Chenzhong Liao Yajun duan Department of Cardiology the First Affiliated Hospital of USTC Division of Life Sciences and Medicine University of Science and Technology of China Key Laboratory of Metabolism and Regulation for Major Diseases of Anhui Higher Education Institutes Hefei University of Technology

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases. The loss of dopaminergic (DAergic) neurons in the substantia nigra and the decrease of dopamine (DA) levels accelerate the process of PD. L-Ergothioneine (EGT) is a natural antioxidant derived from microorganisms， especially in edible mushrooms. EGT can penetrate blood-brain barrier and its levels are significantly decreased in the plasma of PD patients. Therefore， we speculated that EGT could ameliorate PD， and determined its effect on PD development by using 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine (MPTP)-induced PD mouse models and neurotoxin 1-methyl-4-phenylpyridinium (MPP+)-induced cell models. Our results show that EGT alleviated MPTP-induced behavioral dysfunction in mice. Mechanistically， we innovatively revealed that EGT is a key regulator of DJ-1. EGT restored DA levels by activating the DJ-1-Nuclear receptor-related factor 1(Nurr1) axis. Furthermore， it reduced reactive oxygen species (ROS) levels by regulating the DJ-1-Nuclear factor erythroid 2-related factor 2 (Nrf2) pathway， which inhibited oxidative stress-induced DAergic neuronal apoptosis. Combined treatment with DJ-1-siRNA transfection revealed that blocking DJ-1 reversed EGT upregulated Nurr1 and Nrf2 expression in the nucleus， which significantly decreased the benefits of EGT. Taken together， our study suggests that EGT can ameliorate PD and be considered as a strategy for PD treatment.

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基于超高效液相色谱-串联质谱同时测定婴幼儿配方乳粉中2′-岩藻糖基乳糖和乳糖-N-新四糖

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食品与发酵工业 2024年第12期50卷 327-335页

作者：詹胜群葛城严丽周荣杰丁玉珍段潇潇澳优乳业(中国)有限公司湖南长沙410200

建立超高效液相色谱-串联质谱法(ultra-performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)同时测定婴幼儿配方乳粉中2′-岩藻糖基乳糖(2′-fucosyllactose,2′-FL)和乳糖-N-新四糖(lacto-N-neotetraose,LNnT)。样品经水提取,沉淀法除蛋白、针式水系滤膜净化,以乙腈和10 mmol/L甲酸铵水溶液(含0.1%甲酸,体积分数)为流动相,采用酰胺柱进行梯度洗脱,采用选择反应检测扫描模式(selective reaction monitoring,SRM)测定,外标法定量,稀释法评估溶剂效应、提取后添加法评估基质效应。结果表明,正离子模式下2′-FL和LNnT分别采用加合离子峰[M+NH+4]和[M+H+]较好,梯度条件、流速优化后使得2′-FL和LNnT均能在8 min内完成洗脱,溶剂效应考察发现2′-FL和LNnT存在严重的溶剂效应,但在消除试验中发现在稀释20倍的情形下,50%乙腈甲酸铵水溶液作为净化样液的稀释溶剂可消除影响,线性范围考察发现2′-FL和LNnT线性范围分别为5.0~150.0 ng/mL和2.5~75.0 ng/mL,基质效应考察发现2′-FL和LNnT均不存在基质效应。方法验证表明,2′-FL和LNnT各项精密度正确度参数均满足国标要求,该方法适用于婴幼儿配方乳粉中2′-FL和LNnT的同时测定。

关键词：婴幼儿配方乳粉超高效液相色谱-串联质谱 2′-岩藻糖基乳糖乳糖-N-新四糖

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老年特发性正常压力脑积水患者的姿势稳定性特征

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中国组织工程研究 2025年第6期29卷 1208-1213页

作者：梁晓笑郑洁皎段林茹陈茜张廷宇上海中医药大学康复医学院上海市201203 复旦大学附属华东医院上海市200040

背景:姿势控制能力受损是特发性正常压力脑积水患者发生跌倒、引发继发性损害的重要危险因素,现有研究大多是对患者直线行走时的步态参数进行分析,鲜有对患者在静态与动态活动中的姿势稳定性特征进行分析。目的:分析老年特发性正常压力... 详细信息

背景:姿势控制能力受损是特发性正常压力脑积水患者发生跌倒、引发继发性损害的重要危险因素,现有研究大多是对患者直线行走时的步态参数进行分析,鲜有对患者在静态与动态活动中的姿势稳定性特征进行分析。目的:分析老年特发性正常压力脑积水患者的姿势稳定性特征。方法:选择2022年9月至2023年2月在上海市复旦大学附属华东医院神经外科临床诊断为特发性正常压力脑积水的患者22例作为患者组,同时选择健康陪诊家属18名作为健康对照组。分别采用计时起立-行走测试、多方向性伸展测试、Berg平衡量表和静态平衡功能测试(反应时间、移动速度、方向控制、最大偏移距离、端点行程)评估受试者的姿势稳定性特征。结果与结论:①患者组完成计时起立-行走测试所需时间及静态平衡功能测试的反应时间均长于健康对照组(P<0.05),前、后、左、右方向上的伸展测试手臂伸展距离均小于健康对照组(P<0.05),Berg平衡量表得分低于健康对照组(P<0.05),静态平衡功能测试的移动速度、方向控制、最大偏移距离和端点行程均小于健康对照组(P<0.05)。②结果提示,特发性正常压力脑积水患者表现出整体的姿势控制能力障碍,反应能力与执行能力受损使得这类患者在面对来自内部或外部的干扰时无法做出及时、准确的运动反应而出现姿势不稳,增加了发生跌倒的风险。

关键词：特发性正常压力脑积水姿势控制姿势稳定性跌倒风险平衡功能

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铜基气凝胶的制备及其性能研究

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化工新型材料 2024年第S1期52卷 152-156,162页

作者：段建国严新帅王亚雄苏庆宗黄孝笑内蒙古科技大学机械工程学院包头014010 内蒙古科技大学化学与化工学院包头014010 内蒙古自治区煤化工与煤炭综合利用重点实验室包头014010

采用无机分散溶胶-凝胶法,以CuCl_(2)·2H_(2)O作为铜源并辅以CO_(2)超临界干燥和高温处理制备了铜基气凝胶。利用扫描电镜(SEM)、X射线衍射(XRD)、红外光谱(FT-IR)、物理吸附以及热重(TG-DTA)等表征手段,对其在不同处理温度下的微观形... 详细信息

采用无机分散溶胶-凝胶法,以CuCl_(2)·2H_(2)O作为铜源并辅以CO_(2)超临界干燥和高温处理制备了铜基气凝胶。利用扫描电镜(SEM)、X射线衍射(XRD)、红外光谱(FT-IR)、物理吸附以及热重(TG-DTA)等表征手段,对其在不同处理温度下的微观形貌、孔结构和热性能进行研究分析。结果表明:在处理温度200℃下,铜基气凝胶互联网络骨架密实,具有较好的支撑结构。在400℃时样品的主要成分为氧化铜;其比表面积最高可达306.16m^(2)/g。与400℃时的样品相比,室温样品的孔体积增加了427倍,比表面积增大了314倍左右。热分解过程分3个阶段,其中在第二阶段(195.04~317.15℃)样品质量损失量最大,此阶段样品质量减少了29.63%,最终在789.30℃左右热分解达到稳定。

关键词：铜基气凝胶无机分散溶胶-凝胶法热处理孔结构热分解

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Gut microbial profiles and the role in lipid metabolism in Shaziling pigs

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Animal Nutrition 2022年第2期9卷 345-356页

作者： Jie Ma Yehui duan Rui Li xiaoxiao Liang Tiejun Li Xingguo Huang Yulong Yin Jie Yin Animal Nutritional Genome and Germplasm InnovationResearch Center College of Animal Science and TechnologyHunan Agricultural UniversityChangsha410128China Institute of Subtropical Agriculture Chinese Academy of SciencesChangsha410125China Henan Ground Biological Science&Technology Co. Ltd.China

Shaziling pig,a Chinese indigenous breed,has been classified as a fatty pig model.However,the gut microbial development and role in lipid metabolism in Shaziling pigs has been rarely reported.Here,we compared the lipid metabolic and microbial profiles at 30,60,90,150,210,and 300 d of age between Shaziling and Yorkshire pigs.Predictably,there were marked differences in the liver lipids(i.e.,cholesterol,glucose,and low-density lipoprotein)and the lipid related expressions(i.e.,SREBP1/2,LXRa/b,DGAT1/2,and FABP1-3)between Shaziling and Yorkshire pigs.Bacteria sequencing in the ileal digesta and mucosa showed that Shaziling pigs had a higher a-diversity and higher abundances of probiotics,such as Lactobacillus johnsonii,Lactobacillus amylovorus,and Clostridium butyricum.Thirty-five differentiated metabolites were further identified in the mucosa between Shaziling and Yorkshire pigs,which were enriched in the carbohydrate,protein,glucose and amino acid metabolism and bile acid biosynthesis.Furthermore,7 differentiated microbial species were markedly correlated with metabolites,indicating the role of gut microbiota in the host metabolism.Next,the role of differentiated L.johnsonii in lipid metabolism was validated in Duroc×Landrace×Yorkshire(DLY)pigs and the results showed that L.johnsonii mono-colonization promoted lipid deposition and metabolism by altering gut microbiota(i.e.,Megasphaera elsdenii and L.johnsonii)and DGAT1/DGAT2/CD36-PPARg gene expressions.In conclusion,Shaziling pigs exhibited different metabolic and microbial profiles compared with Yorkshire pigs,which might have contributed to the diverse metabolic phenotypes,and the significant enrichment of L.johnsonii in Shaziling pigs promoted lipid metabolism and obesity of DLY pigs,which provided a novel idea to improve the fat content of lean pigs.

关键词： Shaziling pig Gut microbiota Lipid metabolism Lactobacillus johnsonii Fatty acid

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