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Rapid characterization of non-volatile phenolic compounds reveals the reliable chemical markers for authentication of traditional Chinese medicine Xiang-ru among confusing Elsholtzia species

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Chinese Journal of Natural Medicines 2024年第4期22卷 375-384页

作者： ZENG zhen ZHANG Chen HU Jiadong WANG Feiyan WU Ziding WANG Jing ZHANG Jun YANG Shuda CHEN Junfeng LI mingming TONG Qi QIU Shi CHEN Wansheng The SATCM Key Laboratory for New Resources&Quality Evaluation of Chinese Medicine Institute of Chinese Materia MedicaShanghai University of Traditional Chinese MedicineShanghai 201203China Department of Pharmacy Second Affiliated Hospital of Navy Medical UniversityShanghai 200003China Kunming Plant Classification Biotechnology Co. Ltd.Kunming 650500China School of Pharmaceutical Sciences and Yunnan Key Laboratory of Pharmacology for Natural Products Kunming Medical UniversityKunming 650500China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs Beijing100700China

The aerial parts of Mosla chinensis Maxim.and Mosla chinensis cv.'Jiangxiangru'(MCJ)are widely utilized in traditional Chinese medicine(TCM),known collectively as Xiang-ru.However,due to clinical effectiveness concerns and frequent misidentification,the original plants have increasingly been substituted by various species within the genera Elsholtzia and Mosla.The challenge in distinguishing between these genera arises from their similar morphological and metabolic profiles.To address this issue,our study introduced a rapid method for metabolic characterization,employing high-resolution mass spectrometry-based metabolomics.Through detailed biosynthetic and chemometric analyses,we pinpointed five phenolic compounds—salviaflaside,cynaroside,scutellarein-7-O-D-glucoside,rutin,and vicenin-2—among 203 identified compounds,as reliable chemical markers for distinguishing Xiang-ru from closely related Elsholtzia species.This methodology holds promise for broad application in the analysis of plant aerial parts,especially in verifying the authenticity of aromatic traditional medicinal plants.Our findings underscore the importance of non-volatile compounds as dependable chemical markers in the authentication process of aromatic traditional medicinal plants.

关键词： Genus Elsholtzia Genus Mosla Traditional Chinese medicine Xiang-ru Non-volatile phenolic compounds Liquid chromatography-mass spectrometry

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Application of mannose-modified fullerene immunomodulator selectively polarizes tumor-associated macrophages potentiating antitumor immunity

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Nano Research 2023年第11期16卷 12855-12863页

作者： Haoyu Wang Lei Li Xinran Cao mingming zhen Chunru Wang Chunli Bai Beijing National Laboratory for Molecular Sciences Key Laboratory of Molecular Nanostructure and NanotechnologyCAS Research/Education Center for Excellence in Molecular SciencesInstitution Institute of ChemistryChinese Academy of SciencesBeijing 100190China University of Chinese Academy of Sciences Beijing 100049China

Polarization of tumor associated macrophages(TAMs)has been a promising therapeutic paradigm for tumor.However,how to achieve precise regulation of TAMs and high efficiency of tumor immunotherapy is still a huge challenge.Here,we report dicarboxy fullerene modified with mannose(DCFM)as an immunomodulator to selectively polarize TAMs and prominently boost anti-tumor immunity.The dicarboxy fullerene molecule was synthesized through the Prato reaction and further covalently bonded with mannose,obtaining the DCFM with well-defined structure.Due to the exist of mannose in DCFM,it could accurately recognize mannose receptor in TAMs.Our cellular experiment results showed that mannose modification could notably promote the uptake of DCFM by the immunosuppressive M2-type macrophages that effectively reprogrammed M2-type macrophages into anti-tumor M1-type macrophages,leading to enhance the phagocytosis of tumor cells by macrophages and inhibiting tumor cells migration.Subsequently,we observed that DCFM could significantly distribute into tumor tissues by in vivo fluorescence imaging.Importantly,DCFM exhibited a superior anti-tumor efficiency in the subcutaneous colorectal tumor model.In addition,it showed that DCFM precisely polarized TAMs into M1-type macrophages and actively increased the infiltration of cytotoxic T lymphocytes(CTLs),inducing profound tumor growth inhibition.

关键词： fullerene mannose macrophage targeting tumor-associated macrophage polarization cancer immunotherapy

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Mass spectrometry analysis of intact protein N-glycosylation signatures of cells and sera in pancreatic adenocarcinomas

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2024年第1期25卷 51-64页

作者： mingming XU Zhaoliang LIU Wenhua HU Ying HAN zhen WU Sufeng CHEN Peng XIA Jing DU Xumin ZHANG Piliang HAO Jun XIA Shuang YANG Center for Clinical Mass Spectrometry College of Pharmaceutical SciencesSoochow UniversitySuzhou215123China School of Life Science and Technology ShanghaiTech UniversityShanghai201210China State Key Laboratory of Genetic Engineering Department of BiochemistrySchool of Life SciencesFudan UniversityShanghai200438China Department of Clinical Laboratory Center Zhejiang Provincial People’s HospitalPeople’s Hospital of Hangzhou Medical CollegeHangzhou310014China

Pancreatic cancer is among the most malignant cancers,and thus early intervention is the key to better survival outcomes.However,no methods have been derived that can reliably identify early precursors of development into malignancy.Therefore,it is urgent to discover early molecular changes during pancreatic tumorigenesis.As aberrant glycosylation is closely associated with cancer progression,numerous efforts have been made to mine glycosylation changes as biomarkers for diagnosis;however,detailed glycoproteomic information,especially site-specific N-glycosylation changes in pancreatic cancer with and without drug treatment,needs to be further explored.Herein,we used comprehensive solid-phase chemoenzymatic glycoproteomics to analyze glycans,glycosites,and intact glycopeptides in pancreatic cancer cells and patient sera.The profiling of N-glycans in cancer cells revealed an increase in the secreted glycoproteins from the primary tumor of MIA PaCa-2 cells,whereas human sera,which contain many secreted glycoproteins,had significant changes of glycans at their specific glycosites.These results indicated the potential role for tumor-specific glycosylation as disease biomarkers.We also found that AMG-510,a small molecule inhibitor against Kirsten rat sarcoma viral oncogene homolog(KRAS)G12C mutation,profoundly reduced the glycosylation level in MIA PaCa-2 cells,suggesting that KRAS plays a role in the cellular glycosylation process,and thus glycosylation inhibition contributes to the anti-tumor effect of AMG-510.

关键词： Pancreatic cancer Glycosylation Biomarker Glycoproteomics Mass spectrometry

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Functionalized gadofullerene ameliorates impaired glycolipid metabolism in type 2 diabetic mice

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Journal of Genetics and Genomics 2022年第4期49卷 364-376页

作者： Jin Wu Yingbo Chen Xue Li Liyuan Ran Xiangdong Liu Xiaoshuang Wang mingming zhen Shanshan Shao Li Zeng Chunru Wang Bin Liang Jiajun Zhao Yingjie Wu Shandong Provincial Hospital Shandong Laboratory Animal CenterScience and Technology Innovation CenterShandong First Medical University&Shandong Academy of Medical SciencesJinanShandong 250021China Institute for Genome Engineered Animal Models of Human Diseases Dalian Medical UniversityDalianLiaoning 116044China Beijing National Laboratory for Molecular Sciences Key Laboratory of Molecular Nanostructure and NanotechnologyCAS Research/Education Center for Excellence in Molecular SciencesInstitute of ChemistryChinese Academy of SciencesBeijing 100190China Center for Life Sciences School of Life SciencesYunnan UniversityKunmingYunnan 650091China Department of Molecular Pathobiology New York University College of DentistryNew York 10010USA

The soaring global prevalence of diabetes makes it urgent to explore new drugs with high efficacy and safety.Nanomaterial-derived bioactive agents are emerging as one of the most promising candidates for biomedical application.In the present study,we investigated the anti-diabetic effects of a functionalized gadofullerene(GF)using obese db/db and non-obese mouse model of type 2 diabete mellitus(MKR)mouse type 2 diabetes mellitus(T2DM)models.In both mouse models,the diabetic phenotypes,including hyperglycemia,impaired glucose tolerance,and insulin sensitivity,were ameliorated after two or four weeks of intraperitoneal administration of GF.GF lowered blood glucose levels in a dose-dependent manner.Importantly,the restored blood glucose levels could persist ten days after withdrawal of GF treatment.The hepatic AKT/GSK3β/FoxO1 pathway is shown to be the main target of GF for rebalancing gluconeogenesis and glycogen synthesis in vivo and in vitro.Furthermore,GF treatment significantly reduced weight gain of db/db mice with reduced hepatic fat storage by the inhibition of de novo lipogenesis through m TOR/S6K/SREBP1 pathway.Our data provide compelling evidence to support the promising application of GF for the treatment of T2DM.

关键词： Functionalized gadofullerene Type 2 diabetes Glycolipid metabolism AKT De novo lipogenesis

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Oral nano-antioxidants improve sleep by restoring intestinal barrier integrity and preventing systemic inflammation

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National Science Review 2023年第12期10卷 70-82页

作者： Zhanfeng Wu Lei Liu Lei Li Xinran Cao Wang Jia Xiaodan Liao Zhongpu Zhao Hedong Qi Guoqiang Fan Huiqiang Lu Chunying Shu mingming zhen Chunru Wang Chunli Bai Beijing National Laboratory for Molecular Sciences Key Laboratory of Molecular Nanostructure and Nanotechnology CAS ResearchEducation Center for Excellence in Molecular Sciences Institute of Chemistry Chinese Academy of Sciences University of Chinese Academy of Sciences School of Pharmacy Wenzhou Medical University Center for Drug Screening and Research School of Geography and Environmental Engineering Gannan Normal University

Sleep deprivation(SD) is a severe public health threat that can cause systemic inflammation and nerve damage. Few effective and side-effect-free drugs are available to address SD. However, the bidirectional communications between the brain and gut provide new strategies for anti-SD therapeutics. Here we explored oral delivery of fullerene nano-antioxidants(FNAO) in the SD model to improve sleep by regulating abnormal intestinal barrier and systemic inflammation via the brain-gut axis. SD caused excessive reactive oxygen species(ROS) production and hyperactive inflammatory responses in the intestines of zebrafish and mouse models, leading to disturbed sleep patterns and reduced brain nerve activity. Of note,based on the property of the conjugated π bond of the C60structure to absorb unpaired electrons, oral FNAO efficiently reduced the excessive ROS in the intestines, maintained redox homeostasis and intestinal barrier integrity, and ameliorated intestinal and systemic inflammation, resulting in superior sleep improvement. Our findings suggest that maintaining intestinal homeostasis may be a promising avenue for SD-related nerve injury therapy.

关键词： sleep deprivation fullerene nano-antioxidants brain-gut axis reactive oxygen species inflammation

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Nanosize aminated fullerene for autophagic flux activation and G0/G1 phase arrest in cancer cells via post-transcriptional regulation

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Nano Research 2022年第4期15卷 3346-3355页

作者： Xiaoyan Zhang Wei Zhou Yang Liu Linyu Jin Jiawei Huo Yang Yang Shumu Li Haijun Ma Jiao Li mingming zhen Jie Li Chunru Wang Beijing National Research Center for Molecular Sciences Key Laboratory of Molecular Nanostructure and NanotechnologyInstitute of ChemistryChinese Academy of ScienceBeijing 100190China Department of Spinal Surgery Peking University People’s HospitalPeking UniversityBeijing 100034China

Functional fullerene derivatives exhibit special inhibitory effects on tumor progress and metastasis via diverse tumor microenvironment regulations,while the elusive molecular mechanisms hinder their clinical transformation.Herein,it is initially revealed that nanosize aminated fullerene(C_(70)-EDA)can activate autophagic flux,induce G0/G1 cell cycle arrest to abrogate cancer cell proliferation,and significantly inhibit tumor growth in vivo.Mechanismly,C_(70)-EDA promotes the expression of cathepsin D involved in autophagic activation via post-transcriptional regulation,attributing to the interaction with a panel of RNA binding proteins.The accumulation of cathepsin D induces the autophagic degradation of cyclin D1,which arouses G0/G1 phase arrest.This work unveils the fantastic anti-tumor activity of aminated fullerene,elucidates the molecular mechanism,and provides a new strategy for the antineoplastic drug development on functional fullerenes.

关键词： aminated fullerene autophagic flux G0/G1 phase arrest post-transcription regulation

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海马Chrnb4在老龄小鼠术后谵妄中的作用

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中华麻醉学杂志 2024年第6期44卷 682-687页

作者：彭姮玥李靓静张明明何晓帆王臻赵晓春中国医科大学附属口腔医院麻醉科沈阳110002

目的:评价海马含β4亚基的烟碱型乙酰胆碱受体(Chrnb4)在老龄小鼠术后谵妄中的作用。方法:SPF级雄性C56BL/6J小鼠48只,18月龄,体质量29~35 g。采用随机数字表法分为6组:胫骨骨折手术组(TF组, n=6)、假手术组(Sham组, n=6)、胫骨骨折手术... 详细信息

目的:评价海马含β4亚基的烟碱型乙酰胆碱受体(Chrnb4)在老龄小鼠术后谵妄中的作用。方法:SPF级雄性C56BL/6J小鼠48只,18月龄,体质量29~35 g。采用随机数字表法分为6组:胫骨骨折手术组(TF组, n=6)、假手术组(Sham组, n=6)、胫骨骨折手术+阿地芬宁组(TA组, n=9)、胫骨骨折手术+对照溶剂组(TV组, n=9)、假手术+阿地芬宁组(SA组, n=9)和假手术+对照溶剂组(SV组, n=9)。采用七氟烷麻醉下胫骨骨折手术制备术后谵妄模型。假手术为仅进行麻醉和皮肤切口并缝合。TA组、TV组、SA组、SV组于术前5 d置入脑室微量给药套管,每组再随机选取3只小鼠,置入海马CA1区微丝阵列电极。于术后30 min时,TA组和SA组脑室输注阿地芬宁(62.5 nmol/μl)2 μl,TV组和SV组给予对照溶剂2 μl,间断24 h给药,连续给予7 d。TF组及Sham组于术后24 h时,采用实时定量PCR法检测海马组织Chrnb4 mRNA表达水平。TA组、TV组、SA组、SV组分别于术后第7、8天采用旷场实验及O迷宫实验评价冲动样行为;行为学实验结束后,采用免疫荧光染色法检测海马CA1区胶质纤维酸性蛋白(GFAP)及囊泡谷氨酸转运蛋白1(Vglut1)的表达,旷场实验期间记录小鼠海马CA1区局部场电位。结果:与Sham组相比,TF组海马Chrnb4 mRNA表达上调( P0.05)。结论:海马Chrnb4可能参与了老龄小鼠术后谵妄的发生机制,该过程可能与抑制神经元兴奋性毒性有关。

关键词：谵妄术后认知并发症老年人受体烟碱

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HPLC法测定二丁酰环磷腺苷钙原料药中的有关物质

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中国医药工业杂志 2024年第1期55卷 101-106页

作者：孙珍徐明明严翠霞郑璐侠邵泓上海市食品药品检验研究院国家药监局治疗类单抗质量控制重点实验室上海201203

建立了HPLC法测定二丁酰环磷腺苷钙(1)原料药中的有关物质,分别为2'-O-单丁酰环磷腺苷钠(3)、N~6-单丁酰环磷腺苷钠(4)、环磷腺苷(5)。采用ODS Hypersil色谱柱(4.6 mm×200 mm,5μm),以0.1 mol/L磷酸二氢钾溶液和甲醇(体积比600∶400)... 详细信息

建立了HPLC法测定二丁酰环磷腺苷钙(1)原料药中的有关物质,分别为2'-O-单丁酰环磷腺苷钠(3)、N~6-单丁酰环磷腺苷钠(4)、环磷腺苷(5)。采用ODS Hypersil色谱柱(4.6 mm×200 mm,5μm),以0.1 mol/L磷酸二氢钾溶液和甲醇(体积比600∶400)为流动相,检测波长为273 nm;柱温为30℃;流速为1.0 mL/min。建立了加校正因子的主成分自身对照法测定3种已知杂质,确定了3、4、5相对主成分的校正因子,分别是1.7、0.9、1.4。结果显示,二丁酰环磷腺苷钠(2)和3、4、5分别在0.1~20、4~20、1~20、0.1~2μg/mL内线性关系良好。3、4、5的平均加样回收率(n=2)分别为101.0%、98.5%、99.7%,精密度RSD(n=6)分别为0.1%、0.2%、0.1%,重复性RSD(n=6)分别为1.5%、1.6%、2.5%。3、4、5和1的检测限分别为0.2、0.1、0.1、0.2 ng。该方法专属性好、准确度和灵敏度高,为1的质量研究提供了参考。

关键词：二丁酰环磷腺苷钙 HPLC 有关物质加校正因子的主成分自身对照法

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Aptamer-induced in-situ growth of acetylcholinesterase-Cu_(3)(PO_(4))_(2)hybrid nanoflowers for electrochemical detection of organophosphorus inhibitors

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Nano Research 2023年第10期16卷 12134-12143页

作者： Limin Yang Linjiao Qu Xiaolong Zhang mingming Li zhen Liu State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology China University of Petroleum(East China)Qingdao 266580China Institute of Biomedical Engineering Shenzhen Bay LaboratoryShenzhen 518055Chin

Enzyme-inorganic hybrid nanoflowers(HNFs)have shown excellent sensing capabilities due to their large specific surface area as well as the simplicity and mildness of the preparation process.However,coupling HNFs to electrodes to fabricate a uniform and controllable enzymatic electrochemical sensing interface remains a challenge.Here,we proposed an aptamer-induced insitu fabrication strategy for preparing an HNF-based electrochemical sensor with ideal performance.Central to this strategy is the introduction of acetylcholinesterase(AChE)-specific binding aptamer(Apt),which induces the in-situ growth of AChE-copper phosphate(Cu_(3)(PO_(4))_(2))HNFs on the surface of carbon paper(CP).In addition,a dense gold nanoparticle(AuNP)layer was electrodeposited on the CP for anchoring Apt and further extending the electroactive surface area.The prepared AChECu_(3)(PO_(4))_(2)HNF/Apt/AuNP/CP biosensor exhibited a wide detection range from 1 to 107 pM for the four organophosphorus inhibitors,including isocarbophos,dichlorvos,methamidophos,and parathion,with detection limits down to 0.016,0.028,0.071,and 0.113 pM,respectively.With the reactivation of pralidoxime chloride,the electrode can still recover 98.1%of the response after five times of repeated use.In real sample detection,the biosensor achieved high recoveries from 96.45%to 100.13%.The detection target may be extendable to other AChE inhibitors(e.g.,drugs for Alzheimer’s disease).This study demonstrates for the first time the feasibility of using aptamers as an inducer to fabricate an electrochemical enzyme sensing interface in-situ.This strategy can be used to fabricate other enzyme-based biosensors and therefore has broader applications.

关键词： in-situ fabrication aptamer enzyme-inorganic hybrid nanoflowers electrochemical biosensor acetylcholinesterase inhibitors

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基于谷氨酸消旋酶基因(murI)构建杀香鱼假单胞菌减毒活疫苗株的染色体-质粒平衡致死表达系统

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微生物学报 2024年第3期64卷 767-779页

作者：张明明叶浩达张朝政王鹏程王克诠严小军陶震浙江海洋大学水产学院浙江舟山316022 宁波大学海洋学院浙江宁波315211

【目的】构建一种基于谷氨酸消旋酶(MurI)基因的染色体-质粒平衡致死系统,用于杀香鱼假单胞菌减毒活疫苗株(Pseudomonas plecoglossicidaΔtssD-1,PpΔtssD-1)中表达外源抗原,为开发多联活疫苗提供新的思路和方法。【方法】利用同源重... 详细信息

【目的】构建一种基于谷氨酸消旋酶(MurI)基因的染色体-质粒平衡致死系统,用于杀香鱼假单胞菌减毒活疫苗株(Pseudomonas plecoglossicidaΔtssD-1,PpΔtssD-1)中表达外源抗原,为开发多联活疫苗提供新的思路和方法。【方法】利用同源重组技术,将亲本株PpΔtssD-1中的murI基因敲除,构建murI基因缺失突变株;将广宿主穿梭质粒pBBR1MCS-2的卡那霉素抗性基因替换为murI基因,构建平衡致死质粒(即无抗性回补质粒);在平衡致死质粒的多克隆位点处插入绿色荧光蛋白以检测外源抗原是否稳定表达,对重组菌株进行生物学特性分析,包括生长曲线、质粒稳定性和外源抗原表达水平。【结果】murI基因缺失株在不含D-谷氨酸的LB培养基上无法生长;无抗性回补株在不含D-谷氨酸的LB培养基上恢复了生长能力,但生长速度低于亲本株;经鉴定外源抗原可在无抗性质粒中稳定表达,并可在荧光显微镜下观察到明显的绿色荧光信号;此外,平衡致死质粒在重组菌株中具有良好的遗传稳定性。【结论】本研究以murI为靶点构建了新型的染色体-质粒平衡致死系统,可在无抗性筛选条件下在PpΔtssD-1中表达外源抗原,为开发多联活疫苗提供了新的策略和方法。

关键词：杀香鱼假单胞菌ΔtssD-1株营养缺陷株平衡致死系统 D-谷氨酸

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