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The use of melittin to enhance transgene expression mediated by recombinant adeno-associated virus serotype 2 vectors both in vitro and in vivo

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Journal of Integrative Medicine 2023年第1期21卷 106-116页

作者： Yi-lin Xie Ji-yao Wang yun He Xiao-min Yu qing-yun zheng Chen Ling Xi-lin Feng Li-qing Zhu State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology(Ministry of Education) School of Life SciencesZhongshan HospitalFudan UniversityShanghai 200438China Department of Clinical Laboratory the First Affiliated Hospital of Wenzhou Medical UniversityWenzhou 325000Zhejiang ProvinceChina Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province Wenzhou 325000Zhejiang ProvinceChina Yantai Fuheng Biological Technology Co. Ltd.Yantai 264006Shandong ProvinceChin

Objective: Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus(rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based r AAV2 vector delivery strategy.Methods: The melittin peptide was inserted into the rAAV2 capsid either in the loop Ⅷ of all viral proteins(VPs) or at the N terminus of VP2. Various r AAV2-gfp or-fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression.Results: rAAV2 vectors with melittin peptide inserted in the loop Ⅷ of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of r AAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity.Conclusion: Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced

关键词： TAAV Melittin Capsid engineering Co-administration Transduction efficiency

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The use of miR122 and its target sequence in adeno-associated virus-mediated trichosanthin gene therapy

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Journal of Integrative Medicine 2021年第6期19卷 515-525页

作者： Gai Ran Xi-lin Feng Yi-lin Xie qing-yun zheng Peng-peng Guo Ming Yang Ying-lu Feng Chen Ling Li-qing Zhu Chen Zhong State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology(Ministry of Education) School of Life SciencesZhongshan HospitalFudan UniversityShanghai 200438China Division of Cellular and Molecular Therapy Department of PediatricsUniversity of FloridaGainesvilleFL 32611USA Department of Traditional Chinese Medicine Chinese People’s Liberation Army 971 HospitalQingdao 266071Shandong ProvinceChina Department of Clinical Laboratory The First Affiliated Hospital of Wenzhou Medical UniversityWenzhou 325000Zhejiang ProvinceChina

Objective:Plant-derived cytotoxic transgene expression,such as trichosanthin(tcs),regulated by recombinant adeno-associated virus(r AAV)vector is a promising cancer gene therapy.However,the cytotoxic transgene can hamper the vector production in the r AAV producer cell line,human embryonic kidney(HEK293)cells.Here,we explored micro RNA-122(miR122)and its target sequence to limit the expression of the cytotoxic gene in the r AAV producer cells.Methods:A miR122 target(122 T)sequence was incorporated into the 30 untranslated region of the tcs c DNA sequence.The firefly luciferase(fluc)transgene was used as an appropriate control.Cell line HEK293-mir122 was generated by the lentiviral vector-mediated genome integration of the mir122 gene in parental HEK293 cells.The effects of miR122 overexpression on cell growth,transgene expression,and r AAV production were determined.Results:The presence of 122 T sequence significantly reduced transgene expression in the miR122-enriched Huh7 cell line(in vitro),fresh human hepatocytes(ex vivo),and mouse liver(in vivo).Also,the normal liver physiology was unaffected by delivery of 122 T sequence by r AAV vectors.Compared with the parental cells,the miR122-overexpressing HEK293-mir122 cell line showed similar cell growth rate and expression of transgene without 122 T,as well as the ability to produce liver-targeting r AAV vectors.Fascinatingly,the yield of r AAV vectors carrying the tcs-122 T gene was increased by 77.7-fold in HEK293-mir122 cells.Moreover,the tcs-122 T-containing r AAV vectors significantly reduced the proliferation of hepatocellular carcinoma cells without affecting the normal liver cells.Conclusion:HEK293-mir122 cells along with the 122 T sequence provide a potential tool to attenuate the cytotoxic transgene expression,such as tcs,during r AAV vector production.

关键词： Trichosanthin miR122 target sequence Recombinant adeno-associated virus vector Gene therapy Liver cancer

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A Minimum Residual Based Gradient Iterative Method for a Class of Matrix Equations

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Acta Mathematicae Applicatae Sinica 2024年第1期40卷 17-34页

作者： qing-qing zheng Department of Mathematics College of ScienceChina University of Petroleum-BeijingBeijing102249China

In this paper, we present a minimum residual based gradient iterative method for solving a class of matrix equations including Sylvester matrix equations and general coupled matrix equations. The iterative method uses a negative gradient as steepest direction and seeks for an optimal step size to minimize the residual norm of next iterate. It is shown that the iterative sequence converges unconditionally to the exact solution for any initial guess and that the norm of the residual matrix and error matrix decrease monotonically. Numerical tests are presented to show the efficiency of the proposed method and confirm the theoretical results.

关键词： Sylvester matrix equation coupled matrix equation minimum residual gradient descent convergence analysis

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RNA modifications in cellular metabolism:implications for metabolism-targeted therapy and immunotherapy

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Signal Transduction and Targeted Therapy 2024年第4期9卷 1565-1594页

作者： Wei-Wei Liu Si-qing zheng Tian Li yun-Fei Fei Chen Wang Shuang Zhang Fei Wang Guan-Min Jiang Hao Wang Department of Laboratory Medicine The First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiChina School of Clinical Medicine Shandong UniversityJinanChina Core Unit of National Clinical Research Center for Laboratory Medicine HefeiChina Neurosurgical Department The First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiChina Department of Clinical Laboratory The Fifth Affiliated HospitalSun Yat-sen UniversityZhuhaiChina

Cellular metabolism is an intricate network satisfying bioenergetic and biosynthesis requirements of cells.Relevant studies have been constantly making inroads in our understanding of pathophysiology,and inspiring development of therapeutics.As a crucial component of epigenetics at post-transcription level,RNA modification significantly determines RNA fates,further affecting various biological processes and cellular phenotypes.To be noted,immunometabolism defines the metabolic alterations occur on immune cells in different stages and immunological contexts.In this review,we characterize the distribution features,modifying mechanisms and biological functions of 8 RNA modifications,including N6-methyladenosine(m6A),N6,2'-O-dimethyladenosine(m6Am),N1-methyladenosine(m1A),5-methylcytosine(m5C),N4-acetylcytosine(ac4C),N7-methylguanosine(m7G),Pseudouridine(V),adenosine-to-inosine(A-to-l)editing,which are relatively the most studied types.Then regulatory roles of these RNA modification on metabolism in diverse health and disease contexts are comprehensively described,categorized as glucose,lipid,amino acid,and mitochondrial metabolism.And we highlight the regulation of RNA modifications on immunometabolism,further influencing immune responses.Above all,we provide a thorough discussion about clinical implications of RNA modification in metabolism-targeted therapy and immunotherapy,progression of RNA modification-targeted agents,and its potential in RNAtargeted therapeutics.Eventually,we give legitimate perspectives for future researches in this field from methodological requirements,mechanistic insights,to therapeutic applications.

关键词： metabolism targeted alterations

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Multi-time Collaborative Restoration for Integrated Electrical-gas Distribution Systems Based on Rolling Optimization

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CSEE Journal of Power and Energy Systems 2024年第2期10卷 660-671页

作者： Jingyang yun zheng Yan yun Zhou Peichao Zhang Weidong Hu Key Laboratory of Control of Power Transmission and Conversion of Ministry of Education Department of Electrical EngineeringShanghai Jiao Tong University

This paper proposes a multi-time collaborative restoration model for integrated electricity-gas distribution sys-tems(IEGDSs)considering multiple resources after extreme weather events.Based on the linearized power flow constraints of the unbalanced electrical distribution system(EDS)and gas distribution system(GDS),this problem can be formulated as a mixed-integer linear programming(MILP)model.To improve the efficiency and veracity of the solution,a rolling optimiza-tion based two-stage method is developed with the first stage solved by a linear approximation model,and the second stage solved by real-time updated rolling optimization.By solving the MILP problem using rolling optimization,the proposed model and solution method achieve efficient and reliable collaborative restoration of IEGDS considering multiple resources and unbal-anced operation characteristics of EDS.The effectiveness of the proposed model and method is validated by using an IEGDS made of a 37-bus unbalanced EDS and 11-node GDS.Index Terms-Electricity-gas system,mix-integer linear programming,power system restoration.

关键词： Electricity-gas system mix-integer linear programming power system restoration

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Automatic recognition of depression based on audio and video:A review

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World Journal of Psychiatry 2024年第2期14卷 225-233页

作者： Meng-Meng Han Xing-yun Li Xin-Yu Yi yun-Shao zheng Wei-Li Xia Ya-Fei Liu qing-Xiang Wang Shandong Mental Health Center Shandong UniversityJinan 250014Shandong ProvinceChina Key Laboratory of Computing Power Network and Information Security Ministry of EducationShandong Computer Science Center(National Supercomputer Center in Jinan)Qilu University of Technology(Shandong Academy of Sciences)Jinan 250353Shandong ProvinceChina Shandong Engineering Research Center of Big Data Applied Technology Faculty of Computer Science and TechnologyQilu University of Technology(Shandong Academy of Sciences)Jinan 250353Shandong ProvinceChina Shandong Provincial Key Laboratory of Computer Networks Shandong Fundamental Research Center for Computer ScienceJinan 250353Shandong ProvinceChina Department of Ward Two Shandong Mental Health CenterShandong UniversityJinan 250014Shandong ProvinceChina

Depression is a common mental health disorder.With current depression detection methods,specialized physicians often engage in conversations and physiological examinations based on standardized scales as auxiliary measures for depression assessment.Non-biological markers-typically classified as verbal or non-verbal and deemed crucial evaluation criteria for depression-have not been effectively utilized.Specialized physicians usually require extensive training and experience to capture changes in these features.Advancements in deep learning technology have provided technical support for capturing non-biological markers.Several researchers have proposed automatic depression estimation(ADE)systems based on sounds and videos to assist physicians in capturing these features and conducting depression screening.This article summarizes commonly used public datasets and recent research on audio-and video-based ADE based on three perspectives:Datasets,deficiencies in existing research,and future development directions.

关键词： Depression recognition Deep learning Automatic depression estimation System Audio processing Image processing Feature fusion Future development

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Applications of mercury stable isotopes for tracing volcanism in the geologic record

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Science China Earth Sciences 2024年第5期67卷 1436-1458页

作者： qing GONG Ming-Xing LING Wang zheng State Key Laboratory of Nuclear Resources and Environment East China University of TechnologyNanchang 330013China School of Earth System Science Institute of Surface-Earth System ScienceTianjin UniversityTianjin 300072China

Mercury(Hg)enrichments in ancient sediments have been used as a proxy of volcanism,especially large igneous province(LIP)eruptions.However,considering the existence of other potential Hg sources besides volcanoes and the diverse factors(e.g.,organic matters,clay minerals,sulfide minerals and Fe oxides)that can affect Hg sequestration,there are considerable uncertainties to simply regard sedimentary Hg anomalies as signatures of volcanic activities.Mercury stable isotopes,a promising tool for tracing the origins and transformations of Hg,have been increasingly used for determining the causes of Hg spikes and understanding the geochemical behavior of Hg in the geologic record.To date,lots of researches have applied Hg concentrations and Hg isotopes to identify LIP volcanisms linking with significant geological events such as mass extinctions,ocean anoxic events and other environmental perturbations that mainly occurred in the Phanerozoic.However,the results in previous studies clearly show that not all Hg enrichments are derived from volcanic inputs,which emphasize the need for more caution in using Hg as a fingerprint of volcanism.With a better understanding of Hg isotopes in the future,there will be important implications for Hg isotopes to reconstruct volcanic activities in the rock records and their impacts on biological evolution.

关键词： Mercury stable isotopes Large igneous province Mass extinction event Ocean anoxic event

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Plasma spray coating on interconnector toward promoted solid oxide fuel cells and solid oxide electrolysis cells

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Frontiers in Energy 2024年第3期18卷 390-398页

作者： Junwen CAO yun zheng Wenqiang ZHANG Bo YU Institute of Nuclear and New Energy Technology(INET) Collaborative Innovation Center of Advanced Nuclear Energy TechnologyTsinghua UniversityBeijing 100084China Department of Chemical Engineering University of WaterlooWaterlooOntario N2L 3G1Canada Institute of New Energy Materials and Engineering School of Materials Science and EngineeringFuzhou UniversityFuzhou 350108China

Interconnector is a critical component to construct solid oxide cells (SOCs) stack. Oxidation of metallic interconnectors and Cr poisoning caused by oxidation are important factors that lead to long-term performance degradation of SOCs. Coating on the interconnector surface is an important approach to inhibit the oxidation and Cr migration of the interconnector. Herein, (La0.75Sr0.25)0.95MnO3–δ (LSM) and Mn1.5Co1.5O4 (MCO) are used to fabricate the coatings of interconnector. Two advanced thermal spray technology, atmospheric plasma spraying (APS) and low-pressure plasma spray (LPPS), are adopted for the coating preparation. The electrochemical performance, rising and cooling cycle stability, and Cr diffusion inhibition performance of the coatings are tested and evaluated. The result indicates that MCO can generate more uniform and denser coatings than LSM. In addition, MCO coatings prepared by LPPS shows the best electrochemical performance, rising and cooling cycle stability, and Cr diffusion inhibition. The initial area specific resistance (ASR) is 0.0027 Ω·cm2 at 800 °C. After 4 cooling cycle tests, the ASR increases to 0.0032 Ω·cm2 but lower than other samples. Meanwhile, the relative intense of Cr at the interface of SUS430 with MCO coatings fabricated by LPPS is lower than that of MCO fabricated by APS after 4 rising and cooling cycle operations, showing more favorable Cr diffusion inhibition performance.

关键词： interconnector coating plasma spray electrochemical performance Cr diffusion inhibition solid oxide cells(SOCs)

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Proposing a machine learning approach to analyze and predict basic high-temperature properties of iron ore fines and its factors

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Journal of Iron and Steel Research(International) 2024年第5期31卷 1082-1094页

作者： qing-ke Sun Yao-zu Wang Jian-liang Zhang zheng-jian Liu Le-le Niu Chang-dong Shan yun-fei Ma Institute of Artificial Intelligence University of Science and Technology BeijingBeijing100083China School of Intelligence Science and Technology University of Science and Technology BeijingBeijing100083China School of Metallurgical and Ecological Engineering University of Science and Technology BeijingBeijing100083China

The basic high-temperature properties of iron ore play a crucial role in optimizing sintering and ore blending,but the testing process for these properties is complex and has significant lag time,which cannot meet the actual needs of ore blending.A prediction model for the basic high-temperature properties of iron ore fines was thus proposed based on a combination of machine learning algorithms and genetic algorithms.First,the prediction accuracy of different machine learning models for the basic high-temperature properties of iron ore fines was compared.Then,a random forest model optimized by genetic algorithms was built,further improving the prediction accuracy of the model.The test results show that the random forest model optimized by genetic algorithms has the highest prediction accuracy for the lowest assimilation temperature and liquid phase fluidity of iron ore,with a determination coefficient of 0.903 for the lowest assimilation temperature and 0.927 for the liquid phase fluidity after optimization.The trained model meets the fluctuation requirements of on-site testing and has been successfully applied to actual production on site.

关键词： Iron ore Basic high-temperature property Machine learning Random forest Genetic algorithm

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VX-509 attenuates the stemness characteristics of colorectal cancer stem-like cells by regulating the epithelial-mesenchymal transition through Nodal/Smad2/3 signaling

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World Journal of Stem Cells 2024年第2期16卷 207-227页

作者： yun Yuan Xu-Fan Zhang Yu-Chen Li Hong-qing Chen Tian Wen Jia-Lian zheng Zi-Yi Zhao Qiong-Ying Hu Department of Laboratory Medicine Hospital of Chengdu University of Traditional Chinese MedicineChengdu 610072Sichuan ProvinceChina Department of Nuclear Medicine Hospital of Chengdu University of Traditional Chinese MedicineChengdu 610072Sichuan ProvinceChina College of Medical Technology Chengdu University of Traditional Chinese MedicineChengdu 611137Sichuan ProvinceChina Department of Hepatology Affiliated Hospital of Liaoning University of Traditional Chinese MedicineShenyang 110032Liaoning ProvinceChina Hospital of Chengdu University of Traditional Chinese Medicine Chengdu 610072Sichuan ProvinceChina Traditional Chinese Medicine Regulating Metabolic Diseases Key Laboratory of Sichuan Province Chengdu 610072Sichuan ProvinceChina

BACKGROUND Colorectal cancer stem cells(CCSCs)are heterogeneous cells that can self-renew and undergo multidirectional differentiation in colorectal cancer(CRC)patients.CCSCs are generally accepted to be important sources of CRC and are responsible for the progression,metastasis,and therapeutic resistance of CRC.Therefore,targeting this specific subpopulation has been recognized as a promising strategy for overcoming CRC.AIM To investigate the effect of VX-509 on CCSCs and elucidate the underlying mechanism.METHODS CCSCs were enriched from CRC cell lines by in conditioned serum-free medium.Western blot,Aldefluor,transwell and tumorigenesis assays were performed to verify the phenotypic characteristics of the CCSCs.The anticancer efficacy of VX-509 was assessed in HCT116 CCSCs and HT29 CCSCs by performing cell viability analysis,colony formation,sphere formation,flow cytometry,and western blotting assessments in vitro and tumor growth,immunohistochemistry and immunofluorescence assessments in vivo.RESULTS Compared with parental cells,sphere cells derived from HCT116 and HT29 cells presented increased expression of stem cell transcription factors and stem cell markers and were more potent at promoting migration and tumori-genesis,demonstrating that the CRC sphere cells displayed CSC features.VX-509 inhibited the tumor malignant biological behavior of CRC-stem-like cells,as indicated by their proliferation,migration and clonality in vitro,and suppressed the tumor of CCSC-derived xenograft tumors in vivo.Besides,VX-509 suppressed the CSC character-istics of CRC-stem-like cells and inhibited the progression of epithelial-mesenchymal transition(EMT)signaling in vitro.Nodal was identified as the regulatory factor of VX-509 on CRC stem-like cells through analyses of differen-tially expressed genes and CSC-related database information.VX-509 markedly downregulated the expression of Nodal and its downstream phosphorylated Smad2/3 to inhibit EMT progression.Moreover,VX-509 reversed the dedifferentiation of CCSCs and inhibit

关键词： Colorectal cancer stem cells Stemness VX-509 Epithelial-mesenchymal transition Nodal

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