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In situ synthesis and unidirectional insertion of membrane proteins in liposome-immobilized silica stationary phase for rapid preparation of microaffinity chromatography

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Acta Pharmaceutica Sinica B 2022年第9期12卷 3682-3693页

作者： Yanqiu Gu Rong Wang Panpan Chen Shengnan Li Xinyi Chai Chun Chen Yue Liu Yan Cao Diya Lv Zhanying Hong Zhenyu Zhu Yifeng Chai yongfang yuan Xiaofei Chen Department of Pharmacy Shanghai Ninth People's HospitalShanghai Jiao Tong UniversitySchool of MedicineShanghai 201999China School of Pharmacy Naval Medical University(Second Military Medical University)Shanghai 200433China Shanghai Key Laboratory for Pharmaceutical Metabolite Research Shanghai 200433China

Cell membrane affinity chromatography has been widely applied in membrane protein(MP)-targeted drug screening and interaction analysis.However,in current methods,the MP sources are derived from cell lines or recombinant protein expression,which are time-consuming for cell culture or purification,and also difficult to ensure the purity and consistent orientation of MPs in the chromatographic stationary phase.In this study,a novel in situ synthesis membrane protein affinity chromatography(iSMAC)method was developed utilizing cell-free protein expression(CFE)and covalent immobilized affinity chromatography,which achieved efficient in situ synthesis and unidirectional insertion of MPs into liposomes in the stationary phase.The advantages of iSMAC are:1)There is no need to culture cells or prepare recombinant proteins;2)Specific and purified MPs with stable and controllable content can be obtained within 2 h;3)MPs maintain the transmembrane structure and a consistent orientation in the chromatographic stationary phase;4)The flexible and personalized construction of cDNAs makes it possible to analyze drug binding sites.iSMAC was successfully applied to screen PDGFRβinhibitors from Salvia miltiorrhiza and Schisandra chinensis.Micro columns prepared by in-situ synthesis maintain satisfactory analysis activity within 72 h.Two new PDGFRβinhibitors,salvianolic acid B and gomisin D,were screened out with KD values of 13.44 and 7.39μmol/L,respectively.In vitro experiments confirmed that the two compounds decreasedα-SMA and collagen Ⅰ mRNA levels raised by TGF-βin HSC-T6 cells through regulating the phosphorylation of p38,AKT and ERK.In vivo,Sal B could also attenuate CCl_(4)-induced liver fibrosis by downregulating PDGFRβdownstream related protein levels.The iSMAC method can be applied to other general MPs,and provides a practical approach for the rapid preparation of MP-immobilized or other biological solid-phase materials.

关键词： Affinity chromatography In situ synthesis of membrane proteins Unidirectional insertion PDGFRβ Drug screening Ligand–protein interaction Antihepatic fibrosis

Development of 3-mercaptopropyltrimethoxysilane(MPTS)-modified bone marrow mononuclear cell membrane chromatography for screening anti-osteoporosis components from Scutellariae Radix

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Acta Pharmaceutica Sinica B 2020年第10期10卷 1856-1865页

作者： Yanqiu Gu Xiao Chen Yao Wang Yue Liu Leyi Zheng Xiaoqun Li Rong Wang Shaozhan Wang Shengnan Li Yifeng Chai Jiacan Su yongfang yuan Xiaofei Chen Department of Pharmacy Shanghai Ninth People’s HospitalShanghai Jiao Tong UniversitySchool of MedicineShanghai 201999China School of Pharmacy Second Military Medical UniversityShanghai 200433China Department of Orthopedic Trauma Shanghai Changhai HospitalSecond Military Medical UniversityShanghai 200433China

Osteoporosis is a bone metabolic disease caused by the imbalance between osteoblasts and osteoclasts due to excess osteoclastogenesis,manifesting in the decrease of bone density and bone strength.Scutellariae Radix shows good anti-osteoporosis activity,but the effective component is still unclear.Cell membrane chromatography(CMC)is a biological affinity chromatography with membrane immobilized on a silica carrier as the stationary phase.It can realize a dynamical simulation of interactions between drugs and receptors on cell membrane,which is suitable for screening active compounds from complex systems.In this study,the components of Scutellariae Radix with potential anti-osteoporosis activity through inhibiting the differentiation from bone marrow mononuclear cells(BMMCs)to osteoclast were screened by a BMMC/CMC analytical system.Firstly,a new3-mercaptopropyltrimethoxysilane(MPTS)-modified BMMC/CMC stationary phase was developed to realize covalent binding with cell membrane fractions.By investigating the retention time(tR)of the positive drug,the life span of the MPTS-modified CMC columns was significantly improved from 3 to 12 days.Secondly,6 components of Scutellariae Radix were screened to show affinity to membrane receptors on BMMCs by a two-dimensional BMMC/CMC-TOFMS analytical system.Among them,tectochrysin demonstrated the best anti-osteoporosis effect in vitro,which has never been reported.We found that tectochrysin could inhibit the differentiation of BMMCs into osteoclasts induced by receptor activator of nuclear factor-κB ligand(RANKL)and macrophage colony-stimulating factor(M-CSF)in a concentration-dependent manner in vitro.In vivo,it significantly reduced the loss of bone trabeculae in ovariectomized mice,and decreased the level of C-terminal cross-linking telopeptides of type 1 collagen(CTX-1),tartrate-resistant acid phosphatase 5 b(TRAP-5 b),interleukin 6(IL-6)in serum.In conclusion,tectochrysin serves as a potential candidate in the treatment of osteoporosis.The proposed twodimensional MPTS-modi

关键词： 3-Mercaptopropyltrimethoxysilane-modifiedsilica Two-dimensional cell membrane chromatography Scutellariae Radix Anti-osteoporosis Tectochrysin

Identifying potential anti-COVID-19 pharmacological components of traditional Chinese medicine Lianhuaqingwen capsule based on human exposure and ACE2 biochromatography screening

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Acta Pharmaceutica Sinica B 2021年第1期11卷 222-236页

作者： Xiaofei Chen Yunlong Wu Chun Chen Yanqiu Gu Chunyan Zhu Suping Wang Jiayun Chen Lei Zhang Lei Lv Guoqing Zhang yongfang yuan Yifeng Chai Mingshe Zhu Caisheng Wu School of Pharmacy Naval Medical UniversityShanghai 200433China Fujian Provincial Key Laboratory of Innovative Drug Target Research and State Key Laboratory of Cellular Stress Biology School of Pharmaceutical SciencesXiamen UniversityXiamen 361102China Department of Pharmacy Shanghai Ninth People’s HospitalShanghai Jiao Tong UniversityShanghai 201999China Department of Pharmacy Shanghai Eastern Hepatobiliary Surgery HospitalNaval Medical UniversityShanghai 200438China MassDefect Technologies PrincetonNJ 08540USA

Lianhuaqingwen(LHQW)capsule,a herb medicine product,has been clinically proved to be effective in coronavirus disease 2019(COVID-19)pneumonia treatment.However,human exposure to LHQW components and their pharmacological effects remain largely unknown.Hence,this study aimed to determine human exposure to LHQW components and their anti-COVID-19 pharmacological activities.Analysis of LHQW component profiles in human plasma and urine after repeated therapeutic dosing was conducted using a combination of HRMS and an untargeted data-mining approach,leading to detection of 132 LHQW prototype and metabolite components,which were absorbed via the gastrointestinal tract and formed via biotransformation in human,respectively.Together with data from screening by comprehensive 2 D angiotensin-converting enzyme 2(ACE2)biochromatography,8 components in LHQW that were exposed to human and had potential ACE2 targeting ability were identified for further pharmacodynamic evaluation.Results show that rhein,forsythoside A,forsythoside I,neochlorogenic acid and its isomers exhibited high inhibitory effect on ACE2.For the first time,this study provides chemical and biochemical evidence for exploring molecular mechanisms of therapeutic effects of LHQW capsule for the treatment of COVID-19 patients based on the components exposed to human.It also demonstrates the utility of the human exposure-based approach to identify pharmaceutically active components in Chinese herb medicines.

关键词： Lianhuaqingwen capsule PATBS COVID-19 ACE2 Biochromatography Comprehensive 2D analysis In vivo exposure Surface plasma response Molecular docking

全面质量管理在狂犬病暴露处置门诊消毒管理中的应用价值分析

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中国卫生产业 2024年第4期21卷 69-72页

作者：袁永芳刘端山李瑞华莘县疾病预防控制中心预防接种门诊山东聊城252400 莘县疾病预防控制中心健康教育科山东聊城252400

目的分析全面质量管理在狂犬病暴露处置门诊消毒管理中应用的价值。方法选取2022年1月—2023年12月在莘县疾病预防控制中心狂犬病暴露处置门诊工作的10名医务人员作为研究对象,根据门诊消毒管理方案实施的时间进行分组,2022年1—12月实... 详细信息

目的分析全面质量管理在狂犬病暴露处置门诊消毒管理中应用的价值。方法选取2022年1月—2023年12月在莘县疾病预防控制中心狂犬病暴露处置门诊工作的10名医务人员作为研究对象,根据门诊消毒管理方案实施的时间进行分组,2022年1—12月实施常规管理,设为常规组,2023年1—12月实施全面质量管理,设为研究组,比较两组门诊消毒管理工作质量评分、医务人员工作专业度评分、门诊消毒工作不良事件的发生率。结果研究组门诊消毒管理工作质量、医务人员工作专业度各项评分高于常规组,差异有统计学意义(P均0.05)。结论在狂犬病暴露处置门诊消毒管理中全面质量管理模式应用价值较高,能有效提升门诊消毒管理工作的质量和医务人员的专业度,降低工作中不良事件的发生率。

关键词：全面质量管理狂犬病暴露处置门诊消毒管理

A-DA’D-A Type Pentacyclic Small Molecule Acceptors to Exceed 16.5%Efficiency by Heteroatom Effect at the Outer Side Chain

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Chinese Journal of Chemistry 2023年第15期41卷 1815-1822页

作者： Shaofeng Zhu Changzhou Shi Qingya Wei Can Zhu Jie Ren Jianghua Li Lei Meng yongfang Li Jun yuan Yingping Zou State Key Laboratory of Powder Metallurgy College of Chemistry and Chemical EngineeringCentral South UniversityChangshaHunan 410083China Beiing National Laboratory for Molecular Sciences CAS Key Laboratory of Organic SolidsInstitute of ChemistryChinese Academy of SciencesBeijing 100190China. MOE Key Laboratory of Macromolecular Synthesis and Functionalization International Research Center for X PolymersDepartment of Polymer Science and EngineeringZhejiang UniversityHangzhouZhejiang 310027China

Comprehensive Summary In this work,we adopt a“heteroatom side-chains”modification strategy to modify the thiophene units in A-DA'D-A(acceptor-donor-acceptor’-donor-acceptor)type pentacyclic SMAs(small molecule acceptors),that is,introducing branched alkyl chain at theβ-position of thiophene instead of straight alkyl chain,and then introducing oxygen atom at the third-position on the basis of branched chain.Two new pentacyclic SMAs(BZ4F-EH and BZ4F-OEH)were synthesized,and the influence of the heteroatom side-chains on photoelectric properties of A-DA'D-A type pentacyclic SMAs was systematically studied.Compared with our previously reported BZ4F(Y26),BZ4F-EH shows slightly blue-shifted absorption,while BZ4F-OEH has obvious red-shifted absorption.As a result,BZ4F-OEH-based binary device achieved a high power conversion efficiency(PCE)of 16.56%with a fill factor(FF)of 79.3%,which is the highest efficiency of pentacyclic SMAs to date.

关键词： Organic solar cells Small molecule acceptors A-DA'D-A type structure Heteroatom side chains Substituent effects

Therapeutic application of exosomes in ischaemic stroke

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Stroke & Vascular Neurology 2021年第3期6卷 483-495页

作者： yongfang Li Yaohui Tang Guo-yuan Yang Department of Neurology Ruijin HospitalSchool of medcineShanghai Jiao Tong UniversityShanghaiChina Neuroscience and Neuroengineering Center Medx Research InstituteShanghai Jiao Tong University School of Biomedical EngineeringShanghaiChina

Ischaemic stroke is a leading cause of long-term disability in the world,with limited effective treatments.Increasing evidence demonstrates that exosomes are involved in ischaemic pathology and exhibit restorative therapeutic effects by mediating cell–cell communication.The potential of exosome therapy for ischaemic stroke has been actively investigated in the past decade.In this review,we mainly discuss the current knowledge of therapeutic applications of exosomes from different cell types,different exosomal administration routes,and current advances of exosome tracking and targeting in ischaemic stroke.We also briefly summarised the pathology of ischaemic stroke,exosome biogenesis,exosome profile changes after stroke as well as registered clinical trials of exosome-based therapy.

关键词： ischaemic pathology Therapeutic

A-π-A structured non-fullerene acceptors for stable organic solar cells with efficiency over 17%

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Science China Chemistry 2022年第7期65卷 1374-1382页

作者： Wei Liu Jun yuan Can Zhu Qingya Wei Songting Liang Huotian Zhang Guanhaojie Zheng Yunbin Hu Lei Meng Feng Gao yongfang Li Yingping Zou State Key Laboratory of Powder Metallurgy College of Chemistry and Chemical EngineeringCentral South UniversityChangsha 410083China Beijing National Laboratory for Molecular Sciences CAS Key Laboratory of Organic SolidsInstitute of ChemistryChinese Academy of SciencesBeijing 100190China Biomolecular and Organic Electronics IFMLinköping University58183 LinköpingSweden Shanghai Synchrotron Radiation Facility(SSRF) Zhangjiang LabShanghai Advanced Research InstituteChinese Academy of SciencesShanghai 201204China

With the development of photovoltaic materials, especially the small molecule acceptors(SMAs), organic solar cells(OSCs)have made breakthroughs in power conversion efficiencies(PCEs). However, the stability of high-performance OSCs remains a critical challenge for future technological applications. To tackle the inherent instability of SMA materials under the ambient conditions, much effort has been made to improve OSCs stability, including device modification and new materials design. Here we proposed a new electron acceptor design strategy and developed a “quasi-macromolecule”(QM) with an A-π-A structure,where the functionalized π-bridge is used as a linker between two SMAs(A), to improve the long-term stability without deteriorating device efficiencies. Such type of QMs enables excellent synthetic flexibility to modulate their optical/electrochemical properties, crystallization and aggregation behaviors by changing the A and π units. Moreover, QMs possess a unique long conjugated backbone combining high molecular weight over 3.5 k Da with high purity. Compared with the corresponding SMA BTP-4F-OD(Y6-OD), the devices based on newly synthesized A-π-A type acceptors QM1 and QM2 could exhibit better device stability and more promising PCEs of 17.05% and 16.36%, respectively. This kind of “molecular-framework”(A-π-A)structure provides a new design strategy for developing high-efficiency and-stability photovoltaic materials.

关键词： A-π-A type quasi-macromolecule new molecular design strategy high efficiency and long-term stability organic solar cells

Optimizing side chains on different nitrogen aromatic rings achieving 17% efficiency for organic photovoltaics

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Journal of Energy Chemistry 2022年第2期31卷 173-178页

作者： Zhe Li Can Zhu Jun yuan Liuyang Zhou Wei Liu Xinxin Xia c Juan Hong Honggang Chen Qingya Wei Xinhui Lu yongfang Li Yingping Zou College of Chemistry and Chemical Engineering Central South UniversityChangsha 410000HunanChina CAS Key Laboratory of Organic Solids Institute of Chemistry Chinese Academy of SciencesBeijing 100000China Department of Physics The Chinese University of Hong KongHong Kong 999077China

Balancing charge generation and low energy loss(E_(loss)), especially in the wide spectral absorption region is critical to obtain high-performance organic photovoltaics(OPVs). Therefore, Y11-M and Y11-EB are designed and synthesized through modifying alkyl chains on different nitrogen aromatic rings of the reported non-fullerene acceptor Y11. Although all the molecules have almost similar low band-gap(around 1.30 e V), Y11-M and Y11-EB exhibit wider absorption in 410–870 nm region. Eventually, the conventional devices based on Y11-M and Y11-EB possess more efficient charge generation with low Eloss(around 0.44 e V). In addition, outstanding efficiencies of 16.64% and 17.15% with the fill factor of 76.15% and 74.73% are obtained in PM6:Y11-M and PM6:Y11-EB-based devices, both higher than Y11:PM6. The results highlight the importance of rational alkyl chains optimization, and a good structureproperty relationship is established as well.

关键词： Fused-ring benzotriazole Side chains modification Low band-gap acceptors High-efficiency organic photovoltaics

Click chemistry extracellular vesicle/peptide/chemokine nanocarriers for treating central nervous system injuries

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Acta Pharmaceutica Sinica B 2023年第5期13卷 2202-2218页

作者： Huitong Ruan yongfang Li Cheng Wang Yixu Jiang Yulong Han Yiwei Li Dandan Zheng Jing Ye Gang Chen Guo-yuan Yang Lianfu Deng Ming Guo Xingcai Zhang Yaohui Tang Wenguo Cui Department of Orthopaedics Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint DiseasesShanghai Institute of Traumatology and OrthopaedicsRuijin HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Jiaxing Key Laboratory of Basic Research and Clinical Translation on Orthopedic Biomaterials Department of Orthopaedicsthe Second Affiliated Hospital of Jiaxing UniversityJiaxing 314000China Department of Rehabilitation Medicine Ruijin HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai 200025China Med-X Research Institute and School of Biomedical Engineering Shanghai Jiao Tong UniversityShanghai 200030China School of Engineering Massachusetts Institute of TechnologyCambridgeMA 02139USA School of Engineering and Applied Sciences Harvard UniversityCambridgeMA 02138USA

Central nervous system(CNS)injuries,including stroke,traumatic brain injury,and spinal cord injury,are essential causes of death and long-term disability and are difficult to cure,mainly due to the limited neuron regeneration and the glial scar formation.Herein,we apply extracellular vesicles(EVs)secreted by M2 microglia to improve the differentiation of neural stem cells(NSCs)at the injured site,and simultaneously modify them with the injured vascular targeting peptide(DA7R)and the stem cell recruiting factor(SDF-1)on their surface via copper-free click chemistry to recruit NSCs,inducing their neuronal differentiation,and serving as the nanocarriers at the injured site(Dual-EV).Results prove that the Dual-EV could target human umbilical vascular endothelial cells(HUVECs),recruit NSCs,and promote the neuronal differentiation of NSCs in vitro.Furthermore,10 miRNAs are found to be upregulated in Dual-M2-EVs compared to Dual-M0-EVs via bioinformatic analysis,and further NSC differentiation experiment by flow cytometry reveals that among these miRNAs,miR30b-3p,miR-222-3p,miR-129-5p,and miR-155-5p may exert effect of inducing NSC to differentiate into neurons.In vivo experiments show that Dual-EV nanocarriers achieve improved accumulation in the ischemic area of stroke model mice,potentiate NSCs recruitment,and increase neurogenesis.This work provides new insights for the treatment of neuronal regeneration after CNS injuries as well as endogenous stem cells,and the click chemistry EV/peptide/chemokine and related nanocarriers for improving human health.

关键词： Central nervous system injuries Stroke Neural stem cell Neurogenesis Click chemistry Extracellular vesicles Microglia Targeted delivery