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Fluorine-doped carbon quantum dots with deep-red emission for hypochlorite determination and cancer cell imaging

Chinese Chemical Letters 2024年第6期35卷 433-438页

作者： Boran Cheng Lei Cao Chen Li Fang-Yi Huo Qian-Fang Meng Ganglin Tong Xuan Wu Lin-Lin Bu Lang Rao Shubin Wang Department of Oncology Shenzhen Key Laboratory of Gastrointestinal Cancer Translational ResearchCancer InstitutePeking University Shenzhen HospitalShenzhen-Peking University-Hong Kong University of Science and Technology Medical CenterShenzhen 518036China institute of Biomedical Health Technology and Engineering Shenzhen Bay LaboratoryShenzhen 518132China The State Key Laboratory Breeding Base of Basic Science of Stomatology(Hubei-MOST)&Key Laboratory of Oral biomedicine Ministry of Education School&Hospital of StomatologyWuhan UniversityWuhan 430079China

As a type of new carbon-based nanomaterials,carbon dots(CDs)possess exceptional optical properties,making them highly desirable for use in fluorescent ***,the CDs with deep-red(DR)or near-infrared(NIR)emission have rarely been *** this work,we prepared deep-red emissive fluorine-doped carbon quantum dots(F-CDs)by introducing a precursor simultaneously containing fluorine and *** synergistic effect of nitrogen doping and D-π-A pattern production contributed to the maximum emission of F-CDs at 636 nm with an absolute quantum yield of 36.00%±0.68%.Moreover,we designed an F-CDs-based fluorescence assay to determine the content of hypochlorite(ClO^(-)),with a limit of detection(LOD)as low as 15.4 nmol/L,indicating the high sensitivity of F-CDs to ClO^(-).In real samples,the F-CDs-based fluorescent sensor exhibited excellent sensitivity and selectivity in the detection of ClO^(-),with an error below 2%,suggesting their great potential in daily *** cancer cell imaging,the F-CDs not only demonstrated high sensitivity to ClO^(-)but also exhibited excellent mitochondria targeting,as evidenced by the high Pearson's correlation coefficient(PCC)of 0.93 in colocalization *** work presented here suggests the great potential of replacing commercial dyes with F-CDs for highly specific mitochondria labeling and cell imaging.

关键词： Carbon dots Near-infrared Cell imaging Mitochondria labeling Hypochlorite determination

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Bioorthogonal microglia-inspired mesenchymal stem cell bioengineering system creates livable niches for enhancing ischemic stroke recovery via the hormesis

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Acta Pharmaceutica Sinica B 2024年第3期14卷 1412-1427页

作者： Jianpei Xu Yinzhe Sun Yang You Yuwen Zhang Dan Huang Songlei Zhou Yipu Liu Shiqiang Tong Fenfen Ma Qingxiang Song Chengxiang Dai Suke Li Jigang Lei Zhihua Wang Xiaoling Gao Jun Chen Shanghai Pudong hospital&Department of Pharmaceutics School of PharmacyFudan UniversityShanghai 201203China Key Laboratory of Smart Drug Delivery Ministry of EducationSchool of PharmacyFudan UniversityShanghai 201203China Department of Pharmacology and Chemical Biology State Key Laboratory of Oncogenes and Related GenesShanghai Universities Collaborative Innovation Center for Translational MedicineShanghai Jiao Tong University School of MedicineShanghai 200025China Department of Emergency Shanghai Pudong HospitalFudan University Pudong Medical CenterShanghai 201399China institute of Science and Technology for Brain-Inspired Intelligence&Department of Neurology Fudan UniversityShanghai 201203China Department of Rehabilitation Medicine Huashan HospitalFudan UniversityShanghai 201203China Daxing Research institute University of Science and Technology BeijingBiomedical Industry BaseZhongguancun Science and Technology ParkBeijing 102600China Cellular biomedicine Group Inc. Shanghai 201210China

Mesenchymal stem cells(MSCs)experience substantial viability issues in the stroke infarct region,limiting their therapeutic efficacy and clinical *** levels of deadly reactive oxygen radicals(ROS)and proinflammatory cytokines(PC)in the infarct milieu kill transplanted MSCs,whereas low levels of beneficial ROS and PC stimulate and improve engrafted MSCs’*** on the intrinsic hormesis effects in cellular biology,we built a microglia-inspired MSC bioengineering system to transform detrimental high-level ROS and PC into vitality enhancers for strengthening MSC *** system is achieved by bioorthogonally arming metabolic glycoengineered MSCs with microglial membrane-coated nanoparticles and an antioxidative extracellular protective *** this system,extracellular ROSscavenging and PC-absorbing layers effectively buffer the deleterious effects and establish a microlivable niche at the level of a single MSC for ***,the infarct’s inanimate milieu is transformed at the tissue level into a new living niche to facilitate *** engineered MSCs achieved viability five times higher than natural MSCs at seven days after transplantation and exhibited a superior therapeutic effect for stroke recovery up to 28 *** vitality-augmented system demonstrates the potential to accelerate the clinical translation of MSC treatment and boost stroke recovery.

关键词： Ischemic stroke Mesenchymal stem cell Cell engineering Hormesis effect Bioorthogonal chemistry Microglia Reactive oxygen spices Proinflammatory cytokines

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Naked-eye visualization of lymph nodes using fluorescence nanoprobes in non-human primate-animal models

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Nano Research 2024年第8期17卷 7404-7414页

作者： Xiaoyuan Ji Binbin Chu Xiaofeng Wu Zhiming Xia Airui Jiang Chenyu Wang Zhiming Chen Danni Zhong Qiaolin Wei Bin Song Wanlin Li Yiling Zhong Houyu Wang Fenglin Dong Min Zhou Yao He Suzhou Key Laboratory of Nanotechnology and biomedicine Jiangsu Key Laboratory for Carbon-Based Functional Materials&DevicesInstitute of Functional Nano and Soft Materials(FUNSOM)Soochow UniversitySuzhou 215123China School of Public Health Shanghai Jiao Tong University School of MedicineShanghai 200025China Zhejiang university-university of Edinburgh institute(ZJU-UoE institute) Zhejiang UniversityHaining 314400China Department of Ultrasound the First Affiliated Hospital of Soochow UniversitySuzhou 215006China State Key Laboratory(SKL)of Biobased Transportation Fuel Technology Zhejiang UniversityHangzhou 310027China School of Pharmacy Hangzhou Normal UniversityHangzhou 311121China Macao translational Medicine Center Macao University of Science and TechnologyTaipa 999078MacaoChina Macao institute of Materials Science and Engineering Macao University of Science and TechnologyTaipa 999078MacaoChina

Despite sufficient studies performed in non-primate animal models,there exists scanty information obtained from pilot trials in non-human primate animal models,severely hindering nanomaterials moving from basic research into clinical *** herein present a pioneering demonstration of nanomaterials based optical imaging-guided surgical operation by using macaques as a typical kind of non-human primate-animal ***,taking advantages of strong and stable fluorescence of the small-sized(diameter:~5 nm)silicon-based nanoparticles(SiNPs),lymphatic drainage patterns can be vividly visualized in a real-time manner,and lymph nodes(LN)are able to be sensitively detected and precisely excised from small animal models(e.g.,rats and rabbits)to non-human primate animal models(e.g.,cynomolgus macaque(Macaca fascicularis)and rhesus macaque(Macaca mulatta)).Compared to clinically used invisible near-infrared(NIR)lymphatic tracers(i.e.,indocyanine green(ICG);etc.),we fully indicate that the SiNPs feature unique advantages for naked-eye visible fluorescence-guided surgical operation in long-term *** toxicological analysis in macaque models further provides confirming evidence of favorable biocompatibility of the SiNPs *** expect that our findings would facilitate the translation of nanomaterials from the laboratory to the clinic,especially in the field of cancer treatment.

关键词： fluorescent silicon nanoparticles lymph node real-time naked-eye visualization macaque models

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Circulating tumor DNA and its role in detection, prognosis and therapeutics of hepatocellular carcinoma

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Chinese Journal of Cancer Research 2024年第2期36卷 195-214页

作者： Sana Rashid Yingchuan Sun Umair Ali Khan Saddozai Sikandar Hayyat Muhammad Usman Munir Muhammad Usman Akbar Muhammad Babar Khawar Zhiguang Ren Xinying Ji Malik Ihsan Ullah Khan institute of Molecular Biology and Biotechnology University of LahoreLahore 54590Pakistan Department of Internal Oncology(Section Ⅰ) Xuchang Municipal Central HospitalXuchang 461000China institute of translational Medicine Medical CollegeYangzhou UniversityYangzhou 225001China Australian institute for Bioengineering&Nanotechnology the University of QueenslandBrisbane 4072Australia Gomal Center of Biochemistry and Biotechnology Gomal UniversityDera Ismail Khan 29111Pakistan Applied Molecular Biology and biomedicine Lab Department of ZoologyUniversity of NarowalNarowal Punjab 51600Pakistan Kaifeng Municipal Key Laboratory for Infection and Biosafety Henan International Joint Laboratory of Nuclear Protein RegulationSchool of Basic Medical SciencesHenan UniversityKaifeng 475004China Faculty of Basic Medical Subjects Shu-Qing Medical College of ZhengzhouZhengzhou 450064China Department of Medicine Huaxian County People's HospitalHuaxian 456400China

Hepatocellular carcinoma(HCC) is considered the fifth most prevalent cancer among all types of cancers and has the third most morbidity value. It has the most frequent duplication time and a high recurrence rate. Recently, the most unique technique used is liquid biopsies, which carry many markers;the most prominent is circulating tumor DNA(ctDNA). Varied methods are used to investigate ctDNA, including various forms of polymerase chain reaction(PCR) [emulsion PCR(ePCR), digital PCR(dPCR), and bead, emulsion, amplification, magnetic(BEAMing) PCR]. Hence ctDNA is being recognized as a potential biomarker that permits early cancer detection,treatment monitoring, and predictive data on tumor burden are subjective to therapy or surgery. Numerous ctDNA biomarkers have been investigated based on their alterations such as 1) single nucleotide variations(either insertion or deletion of a nucleotide) markers including TP53, KRAS, and CCND1;2) copy number variations which include markers such as CDK6, EFGR, MYC and BRAF;3) DNA methylation(RASSF1A, SEPT9, KMT2C and CCNA2);4) homozygous mutation includes ctDNA markers as CDKN2A, AXIN1;and 5) gain or loss of function of the genes, particularly for HCC. Various researchers have conducted many studies and gotten fruitful ***, there are some drawbacks to ctDNA namely low quantity, fragment heterogeneity, less stability, limited mutant copies and standards, and differential sensitivity. However, plenty of investigations demonstrate ctDNA's significance as a polyvalent biomarker for cancer and can be viewed as a future diagnostic, prognostic and therapeutic agent. This article overviews many conditions in genetic changes linked to the onset and development of HCC, such as dysregulated signaling pathways, somatic mutations, single-nucleotide polymorphisms, and genomic instability. Additionally, efforts are also made to develop treatments for HCC that are molecularly targeted and to unravel some of the genetic pathways that f

关键词： Hepatocellular carcinoma circulating tumor DNA biomarkers single nucleotide variations diagnosis prognosis

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Supercritical fluid technology:A game-changer for biomacromolecular nanomedicine preparation and biomedical application

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Chinese Chemical Letters 2024年第7期35卷 89-99页

作者： Yating Zheng Yulan Huang Jing Luo Xuqi Peng Xiran Gui Gang Liu Yang Zhang State Key Laboratory of Vaccines for Infectious Diseases Xiang An Biomedicine LaboratoryNational Innovation Platform for Industry-Education Integration in Vaccine ResearchState Key Laboratory of Molecular Vaccinology and Molecular DiagnosticsCenter for Molecular Imaging and Translational MedicineSchool of Public HealthXiamen UniversityXiamen 361102China Shen Zhen Research institute of Xiamen university Shenzhen 518057China Division of Urology and Department of Surgery Brigham and Women's HospitalHarvard Medical SchoolBostonMA 02115United States Innovation Laboratory for Sciences and Technologies of Energy Materials of Fujian Province(IKKEM) Xiamen 361102China

Biomacromolecules are attractive in biomedical applications as therapeutic agents and potential drug carriers due to their natural active components,good biocompatibility,and high ***,their large relative molecular weight,complex structure,susceptibility to degradation,and poor stability limit their *** can address these issues by improving the therapeutic value,bioavailability,permeability,and absorption of biomacromolecules while regulating their retention time in the ***,compelling evidence has been reported that supercritical fluid(SCF)technology has emerged as an alternative that maintains the integrity of biomacromolecules and reduces environmental *** this review,we highlight a set of unique nanosizing strategies based on SCF technology for biomacromolecular nanomedicine,and extensively discuss their characteristics and *** particular,the protein-based,nucleic acid-based,and polysaccharide-based nanomedicine preparations via SCF technology and their biomedical applications are summarized,and the potential for industrial production of biomacromolecular drugs is also *** further provide perspectives on the opportunities and challenges in this excellent field of biomacromolecular drugs nanotechnology.

关键词： Biomacromolecules Supercritical fluid technology Therapeutic proteins Gene therapy Nanomedicine

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Supercritical fluid technology: A game-changer for biomacromolecular nanomedicine preparation and biomedical application

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Chinese Chemical Letters 2024年第7期 115-125页

作者： Yating Zheng Yulan Huang Jing Luo Xuqi Peng Xiran Gui Gang Liu Yang Zhang State Key Laboratory of Vaccines for Infectious Diseases Xiang An Biomedicine LaboratoryNational Innovation Platform for Industry-Education Integration in Vaccine ResearchState Key Laboratory of Molecular Vaccinology and Molecular DiagnosticsCenter for Molecular Imaging and Translational MedicineSchool of Public HealthXiamen University Division of Urology and Department of Surgery Brigham and Women's HospitalHarvard Medical School Innovation Laboratory for Sciences and Technologies of Energy Materials of Fujian Province(IKKEM) Shen Zhen Research institute of Xiamen university

Biomacromolecules are attractive in biomedical applications as therapeutic agents and potential drug carriers due to their natural active components, good biocompatibility, and high targeting. However, their large relative molecular weight, complex structure, susceptibility to degradation, and poor stability limit their usefulness. Nanotechnology can address these issues by improving the therapeutic value, bioavailability, permeability, and absorption of biomacromolecules while regulating their retention time in the body. Especially, compelling evidence has been reported that supercritical fluid(SCF) technology has emerged as an alternative that maintains the integrity of biomacromolecules and reduces environmental contamination. In this review, we highlight a set of unique nanosizing strategies based on SCF technology for biomacromolecular nanomedicine, and extensively discuss their characteristics and *** particular, the protein-based, nucleic acid-based, and polysaccharide-based nanomedicine preparations via SCF technology and their biomedical applications are summarized, and the potential for industrial production of biomacromolecular drugs is also considered. We further provide perspectives on the opportunities and challenges in this excellent field of biomacromolecular drugs nanotechnology.

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Outcomes of combined mitochondria and mesenchymal stem cellsderived exosome therapy in rat acute respiratory distress syndrome and sepsis

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World Journal of Stem Cells 2024年第6期16卷 690-707页

作者： Kun-Chen Lin Wen-Feng Fang Jui-Ning Yeh John Y Chiang Hsin-Ju Chiang Pei-Lin Shao Pei-Hsun Sung Hon-Kan Yip Department of Anesthesiology Kaohsiung Chang Gung Memorial HospitalKaohsiung 83301TaiwanChina Division of Pulmonary and Critical Care Medicine Department of Internal MedicineKaohsiung Chang Gung Memorial HospitalChang Gung University College of MedicineKaohsiung 83301TaiwanChina Department of Cardiology The First Affiliated Hospital of Jinan UniversityGuangzhou 510630Guangdong ProvinceChina Department of Computer Science and Engineering National Sun Yat-sen UniversityKaohsiung 804201TaiwanChina Department of Healthcare Administration and Medical Informatics Kaohsiung Medical UniversityKaohsiung 80708TaiwanChina School of Medicine Chung Shan Medical UniversityTaichung 40201TaiwanChina Department of Nursing Asia UniversityTaichung 41354TaiwanChina Division of Cardiology Department of Internal MedicineKaohsiung Chang Gung Memorial Hospital and Chang Gung University College of MedicineKaohsiung 83301TaiwanChina institute for translational Research in biomedicine Kaohsiung Chang Gung Memorial HospitalKaohsiung 83301TaiwanChina Center for Shockwave Medicine and Tissue Engineering Kaohsiung Chang Gung Memorial HospitalKaohsiung 83301TaiwanChina Department of Medical Research China Medical University HospitalChina Medical UniversityTaichung 40402TaiwanChina School of Medicine College of MedicineChang Gung UniversityTaoyuan 33302TaiwanChina

BACKGROUND The treatment of acute respiratory distress syndrome(ARDS)complicated by sepsis syndrome(SS)remains *** To investigate whether combined adipose-derived mesenchymal-stem-cells(ADMSCs)-derived exosome(EXAD)and exogenous mitochondria(mitoEx)protect the lung from ARDS complicated by *** In vitro study,including L2 cells treated with lipopolysaccharide(LPS)and in vivo study including male-adult-SD rats categorized into groups 1(sham-operated-control),2(ARDS-SS),3(ARDS-SS+EXAD),4(ARDS-SS+mitoEx),and 5(ARDS-SS+EXAD+mitoEx),were included in the present *** In vitro study showed an abundance of mitoEx found in recipient-L2 cells,resulting in significantly higher mitochondrial-cytochrome-C,adenosine triphosphate and relative mitochondrial DNA levels(P<0.001).The protein levels of inflammation[interleukin(IL)-1β/tumor necrosis factor(TNF)-α/nuclear factor-κB/toll-like receptor(TLR)-4/matrix-metalloproteinase(MMP)-9/oxidative-stress(NOX-1/NOX-2)/apoptosis(cleaved-caspase3/cleaved-poly(ADP-ribose)polymerase)]were significantly attenuated in lipopolysaccharide(LPS)-treated L2 cells with EXAD treatment than without EXAD treatment,whereas the protein expressions of cellular junctions[occluding/β-catenin/zonula occludens(ZO)-1/E-cadherin]exhibited an opposite pattern of inflam-mation(all P<0.001).Animals were euthanized by 72 h post-48 h-ARDS induction,and lung tissues were *** 72 h,flow cytometric analysis of bronchoalveolar lavage fluid demonstrated that the levels of inflam-matory cells(Ly6G+/CD14+/CD68+/CD11b/c+/myeloperoxidase+)and albumin were lowest in group 1,highest in group 2,and significantly higher in groups 3 and 4 than in group 5(all P<0.0001),whereas arterial oxygen-saturation(SaO2%)displayed an opposite pattern of albumin among the *** findings of lung injury/fibrosis area and inflammatory/DNA-damaged markers(CD68+/γ-H2AX)displayed an identical pattern of SaO2%among the groups(all P<0.0001).The pro

关键词： Acute respiratory distress syndrome Sepsis syndrome Exosomes Mitochondria Adipose tissue-derived mesenchymal stem cells Inflammation

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Key role of interferon regulatory factor 1(IRF-1)in regulating liver disease:progress and outlook

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Journal of Zhejiang university-Science B(biomedicine & Biotechnology) 2024年第6期25卷 451-470页

作者： Tao CHEN Shipeng LI Dewen DENG Weiye ZHANG Jianjun ZHANG Zhongyang SHEN First Central Clinical School Tianjin Medical UniversityTianjin 300192China Department of Organ Transplant Tianjin First Central HospitalTianjin 300192China Research institute of Transplant Medicine Nankai UniversityTianjin 300192China Department of Hepatobiliary Surgery Henan Provincial People’s HospitalZhengzhou 450000China Key Laboratory of Organ Transplant Tianjin First Central HospitalTianjin 300192China

Interferon regulatory factor 1(IRF-1)is a member of the IRF *** is the first transcription factor to be identified that could bind to the interferon-stimulated response element(ISRE)on the target gene and displays crucial roles in the interferoninduced signals and ***-1,as an important medium,has all of the advantages of full cell cycle regulation,cell death signaling transduction,and reinforcing immune surveillance,which are well *** studies indicate that IRF-1 is of vital importance to the occurrence and evolution of multifarious liver diseases,including but not limited to inhibiting the replication of the hepatitis virus(A/B/C/E),alleviating the progression of liver fibrosis,and aggravating hepatic ischemiareperfusion injury(HIRI).The tumor suppression of IRF-1 is related to the clinical characteristics of liver cancer patients,which makes it a potential indicator for predicting the prognosis and recurrence of liver cancer;additionally,the latest studies have revealed other effects of IRF-1 such as protection against alcoholic/non-alcoholic fatty liver disease(AFLD/NAFLD),cholangiocarcinoma suppression,and uncommon traits in other liver diseases that had previously received little ***,several compounds and drugs have featured a protective function in specific liver disease models in which there is significant involvement of the IRF-1 *** this paper,we hope to propose a prospective research basis upon which to help decipher translational medicine applications of IRF-1 in liver disease treatment.

关键词： Interferon regulatory factor(IRF-1) Hepatitis virus Liver fibrosis Hepatic ischemia-reperfusion injury(HIRI) Liver cancer

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Association between Mental Disorders and Subsequent Medical Conditions

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四川生理科学杂志 2024年第4期46卷 217-217页

作者： Natalie C Momen National Center for Register-based Research School of Business and Social Sciences at Aarhus Uni versity 8210 Aarhus V Denmark. Center for Integrated Register-based Research the Departments of Biomedicine-Human Genetics and Public Health the Center for Integrative Sequencing and the Big Data Center for Environment and Health Aarhus University. Lundbeck Foundation Initiative for Integrative Psychiatric Research the Center for Genomics and Personalized Medicine the Department of Neurology and the Research Unit Department of Psychosis Aarhus University Hospital and the Research Unit for General Practice Aarhus the Copenhagen Research Center for Mental Health Mental Health Center Copenhagen Copenhagen University Hospital Psychiatric Center Copenhagen. Department of Health Care Policy Harvard Medical School Boston the Neurogenomics Division Translational Genomics Research Institute Phoenix AZ the Department of Psychological Medicine Dunedin School of Medicine University of Otago Dunedin New Zealand Sievert Consulting Minneapolis and the Institute for Health Metrics and Evaluation University of Washington Seattle. Faculty of Health and Medical Sciences the Department of Clinical Medicine and the Lundbeck Foundation GeoGenetics Center GLOBE Institute University of Copenhagen Copenhagen and the Institute of Biological Psychiatry Mental Health Center Sankt Hans Mental Health Services Capital Region Roskilde Denmark the National Drug and Alcohol Research Centre University of New South Wales Sydney. Departments of Psychology and Developmental Psychology Heymans Institute and the Interdisciplinary Center Psychopathology and Emotion Regulation University of Groningen Groningen the Netherlands. Department of General Practice Melbourne Medical School University of Melbourne Melbourne VIC and the Queensland Brain Institute Queensland Centre for Mental Health Research the Park Centre for Mental Health and the School of Public Health Faculty of Medicine University of Queensland

Background:Persons with mental disorders are at a higher risk than the general population for the subsequent development of certain medical ***:We used a population-based cohort from Danish national registries that included data on more than 5.9 million persons born in Denmark from 1900 through 2015 and followed them from 2000 through 2016,for a total of 83.9 million person-years.

关键词： conditions milli population

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Bee Sting-Inspired Inflammation-Responsive Microneedles for Periodontal Disease Treatment

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Research 2023年第4期2023卷 23-31页

作者： Chuanhui Song Xiaoxuan Zhang Minhui Lu Yuanjin Zhao Department of Rheumatology and Immunology Institute of Translational MedicineThe Affliated Drum Tower Hospital of Nanjing University Medical SchoolNanjing 210002China State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Chemistry and biomedicine Innovation Center Nanjing UniversityNanjing 210023 China

Periodontal lesions are common and frustrating diseases that impact life *** in this aspect aim at developing local drug delivery systems with better efficacy and less ***,inspired by the sting separation behavior of bees,we conduct novel reactive oxygen species(ROS)-responsive detachable microneedles(MNs)that carry antibiotic metronidazole(Met)for controllable periodontal drug delivery and periodontitis *** from the needle-base separation ability,such MNs can penetrate through the healthy gingival to reach the gingival sulcus's bottom while offering minimal impact to oral ***,as the drug-encapsulated cores were protected by poly(lactic-co-glycolic acid)(PLGA)shells in MNs,the surrounding normal gingival tissue is not affected by Met,resulting in excellent local ***,with the ROS-responsive PLGA-thioketal-polyethylene glycol MN tips,they can be unlocked to release Met directly around the pathogen under the high ROS in the periodontitis sulcus,bringing about improved therapeutic *** on these characteristics,the proposed bioinspired MNs show good therapeutic results in treating a rat model with periodontitis,implying their potential in periodontal disease.

关键词： needle separation bringing

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