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Resuscitation fluids as drugs:targeting the endothelial Glycocalyx

Chinese Medical Journal 2022年第2期135卷 137-144页

作者： Guangjian Wang Hongmin Zhang Dawei Liu Xiaoting Wang Chinese Critical Ultrasound Study Group Department of Critical Care Medicine Peking Union Medical College HospitalPeking Union Medical College&Chinese Academy of Medical SciencesBeijing 100730China

Fluid resuscitation is an essential intervention in critically ill patients,and its ultimate goal is to restore tissue perfusion.Critical illnesses are often accompanied by Glycocalyx degradation caused by inflammatory reactions,hypoperfusion,shock,and so forth,leading to disturbed microcirculatory perfusion and organ dysfunction.Therefore,maintaining or even restoring the Glycocalyx integrity may be of high priority in the therapeutic strategy.Like drugs,however,different resuscitation fluids may have beneficial or harmful effects on the integrity of the Glycocalyx.The purpose of this article is to review the effects of different resuscitation fluids on the Glycocalyx.Many animal studies have shown that normal saline might be associated with Glycocalyx degradation,but clinical studies have not confirmed this finding.Hydroxyethyl starch(HES),rather than other synthetic colloids,may restore the Glycocalyx.However,the use of HES also leads to serious adverse events such as acute kidney injury and bleeding tendencies.Some studies have suggested that albumin may restore the Glycocalyx,whereas others have suggested that balanced crystalloids might aggravate Glycocalyx degradation.Notably,most studies did not correct the effects of the infusion rate or fluid volume;therefore,the results of using balanced crystalloids remain unclear.Moreover,mainly animal studies have suggested that plasma may protect and restore Glycocalyx integrity,and this still requires confirmation by high-quality clinical studies.

关键词： Fluid resuscitation Resuscitation fluid Fluid therapy Endothelial Glycocalyx Glycocalyx

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Endothelial Glycocalyx as a potential theriapeutic target in organ injuries

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Chinese Medical Journal 2019年第8期132卷 963-975页

作者： Rui-Na Cao Li Tang Zhong-Yuan Xia Rui Xia Department of Anesthesiology The First Affiliated Hospital of Yangtze UniversityJingzhouHubei 434000China Department of Anesthesiology Renmin Hospital of Wuhan UniversityWuhanHubei 430061China

Objective:The endothelial Glycocalyx(eGC)is a dynamic and multicomponent layer of macromolecules found at the surface of vascular endothelium,which is largely underappreciated.It has recently been recognized that eGC is a major regulator of endothelial function and may have therapeutic value in organ injuries.This study aimed to explore the role of the eGC in various pathologic and physiologic conditions,by reviewing the basic research findings pertaining to the detection of the eGC and its clinical significance.We also explored different pharmacologic agents used to protect and rebuild the eGC.Data sources:An in-depth search was performed in the PubMed database,focusing on research published after 2003 with keywords including eGC,permeability,Glycocalyx and injuries,and Glycocalyx protection.Study selection:Several authoritative reviews and original studies were identified and reviewed to summarize the characteristics of the eGC under physiologic and pathologic conditions as well as the detection and protection of the eGC.Results:The eGC degradation is closely associated with pathophysiologic changes such as vascular permeability,edema formation,mechanotransduction,and clotting cascade,together with neutrophil and platelet adhesion in diverse injury and disease states including inflammation(sepsis and trauma),ischemia-reperfusion injury,shock,hypervolemia,hypertension,hyperglycemia,and high Na+as well as diabetes and atherosclerosis.Therapeutic strategies for protecting and rebuilding the eGC should be explored through experimental test and clinical verifications.Conclusions:Disturbance of the eGC usually occurs at early stages of various clinical pathophysiologies which can be partly prevented and reversed by protecting and restoring the eGC.The eGC seems to be a promising diagnostic biomarker and therapeutic target in clinical settings.

关键词： Endothelial Glycocalyx Permeability Injuries Glycocalyx protection

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Early diabetic kidney disease:Focus on the Glycocalyx

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World Journal of Diabetes 2023年第5期14卷 460-480页

作者： Hui Yu Yi-Yun Song Xian-Hua Li Department of Nephrology Qilu Hospital of Shandong UniversityJinan 250012Shandong ProvinceChina

The incidence of diabetic kidney disease(DKD)is sharply increasing worldwide.Microalbuminuria is the primary clinical marker used to identify DKD,and its initiating step in diabetes is glomerular endothelial cell dysfunction,particularly Glycocalyx impairment.The Glycocalyx found on the surface of glomerular endothelial cells,is a dynamic hydrated layer structure composed of proteoglycans,glycoproteins,and some adsorbed soluble components.It reinforces the negative charge barrier,transduces the shear stress,and mediates the interaction of blood corpuscles and podocytes with endothelial cells.In the highglucose environment of diabetes,excessive reactive oxygen species and proinflammatory cytokines can damage the endothelial Glycocalyx(EG)both directly and indirectly,which induces the production of microalbuminuria.Further research is required to elucidate the role of the podocyte Glycocalyx,which may,together with endothelial cells,form a line of defense against albumin filtration.Interestingly,recent research has confirmed that the negative charge barrier function of the Glycocalyx found in the glomerular basement membrane and its repulsion effect on albumin is limited.Therefore,to improve the early diagnosis and treatment of DKD,the potential mechanisms of EG degradation must be analyzed and more responsive and controllable targets must be explored.The content of this review will provide insights for future research.

关键词： Glycocalyx Diabetic kidney disease Endothelial cells Reactive oxygen species Microalbuminuria Enzyme

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Effects of Cryopreservation on Human Sperm Glycocalyx

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Reproductive and Developmental Medicine 2017年第4期1卷 233-238页

作者： Yan-Cheng Wu Ai-Jie Xin Hui Lu Hua Diao Li Cheng Yi-Hua Gu Bin Wu Sheng-Ce Tao Zheng Li Hui-Juan Shi Yong-Lian Zhang Department of Applied Biology State Key Laboratory of Bioreactor EngineeringSchool of BiotechnologyEast China University of Science and TechnologyShanghai 200237China Shanghai Key Laboratory of Female Reproductive Endocrine-Related Diseases Shanghai Ji Ai Genetics and IVF InstituteObstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Department of Allergy Renji HospitalShanghai Jiao Tong University School of MedicineShanghai 200001China Department of Reproductive Pharmacology NPFPC Key Laboratory of Contraceptives and DevicesShanghai Institute of Planned Parenthood ResearchShanghai 200032China Shanghai Center for Systems Biomedicine Shanghai Jiao Tong UniversityShanghai 201100China Department of Andrology Urologic Medical CenterShanghai General HospitalShanghai Jiao Tong UniversityShanghai 200080China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

Background:To study the effects of cryopreservation on human sperm Glycocalyx.Methods:The lectin binding profilings of sperm after freeze-thaw were compared by lectin microarray.Results:CryoSperm^(TM) and direct fumigation were confirmed to be the optimized cryoprotectant and method by comparing the sperm recovery rate.In 91 lectins,33 lectins were significantly changed after sperm cryopreservation.Among them,9 lectins greatly decreased and 24 lectins mainly increased.The binding signals of MAA,PSA,ABA,and AIA were verified by FACS,and the results were consistent with that of lectin microarray.Conclusions:Sperm Glycocalyx had significant changes after cryopreservation.The sialic acid,playing an important role in protecting sperm,was greatly lost,which exposed the inner carbohydrates.Thus,the Glycocalyx damage due to the cryopreservation might be one of the reasons for low sperm fertility.

关键词： Cryopreservation Damage Glycocalyx Lectin Microarray Sperm

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Organization and Ultra-Structural Components of Endothelial Surface Glycocalyx Revealed by Stochastic Optical Reconstruction Microscopy(STORM)

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医用生物力学 2019年第A01期34卷 6-7页

作者： Jie Fan Yi Sun Yifan Xia John M.Tarbell Bingmei M.Fua Department of Biomedical Engineering the City College of the City University of New YorkNew YorkNY 10031USA Department of Electrical Engineering the City College of the City University of New YorkNew YorkNY 10031USA

Introduction The endothelial cells(ECs)lining every blood vessel wall constantly expose to the mechanical forces generated by the blood flow.The EC responses to these hemodynamic forces play a critical role in the homeostasis of the circulatory system.In addition to forming a transport barrier between the blood and vessel wall,vascular ECs play important roles in regulating circulation functions.Besides biochemical stimuli,blood flow induced(hemodynamic)mechanical stimuli,such as shear stress,pressure and circumferential stretch,modulate EC morphology and functions by activating mechanosensors,signaling pathways,and gene and protein expressions.The EC responses to the hemodynamic forces(mechano-sensing and transduction)are critical to maintaining normal vascular functions.Failure in the mechano-sensing and transduction leads to serious vascular diseases including hypertension,atherosclerosis,aneurysms and thrombosis,to name a few[1].On the luminal surface of our blood vessels,there is a thin layer called endothelial surface Glycocalyx(ESG)which consists of proteoglycans,glycosaminoglycans(GAGs)and glycoproteins.The GAGs in the ESG are heparan sulfate(HS),hyaluronic acid(HA),chondroitin sulfate(CS),and sialic acid(SA)[2].In order to play important roles in vascular functions,such as being a mechanosensor and transducer for the endothelial cells(ECs)to sense the blood flow,a molecular sieve to maintain normal microvessel permeability and a barrier between the circulating cells and endothelial cells forming the vessel wall,the ESG should have an organized structure at the molecular level.Due to the limitations of optical and electron microscopy,the ultra-structure and organization of ESG has not been revealed until recent development of a super high resolution fluorescence optical microscope,STORM(Stochastic Optical Reconstruction Microscopy).The diffraction of a single fluorescence molecule can be described as the point spread function(PSF).When the light of wavelengthλexcites the fluorophore(emitter),the intensity

关键词： Organization Ultra-Structural Components Endothelial Surface Glycocalyx Revealed Optical Reconstruction Microscopy STORM

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Friction reduction of Glycocalyx of human endothelial cells cultured on anionic hydrogel scaffolds

Friction reduction of glycocalyx of human endothelial cells ...

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2007年全国高分子学术论文报告会

作者：陈咏梅龚建萍 Takayuki Kurokawa Masaru Tanaka Yasuda Kazunori Sadaaki Yamamoto Masatsugu Shimomura Yoshihito Osada Graduate School of Science Hokkaido University Sapporo 060-0810Japan Creative Research Initiative’SOUSEI’Hokkaido UniversitySapporo 001-0021 Graduate School of Science Hokkaido University Sapporo 060-0810Japan Graduate School of Science Hokkaido University Sapporo 060-0810Japan Nanotechnology Research Center Research Institute for Electronic Science Hokkaido UniversitySapporo001-0021Japan Creative Research Initiative’SOUSEI’ Hokkaido University Sapporo 001-0021 School of MedicineHokkaido UniversitySapporo 060-0810Japan Creative Research Initiative’SOUSEI’ Hokkaido University Sapporo 001-0021 Nanotechnology Research Center Research Institute for Electronic Science Hokkaido UniversitySapporo001-0021Japan Graduate School of Science Hokkaido University Sapporo 060-0810Japan

＜正＞In the in vivo vascular system,the inner surface of blood vessel is covered by a monolayer of endothelial cell（EC）that exposes to the blood flow.With great interesting, the red blood cells can deform and smoothly squeeze though the capillary of a diameter much smaller.In this process,the friction should occur between soft ECs and deformable blood red cells.Apparently,only when the surface friction between the inner surface wall of

关键词： hydrogel cell culture Glycocalyx friction platelet adhesion

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Influence of in vitro capacitation time on structural and functional human sperm parameters

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Asian Journal of Andrology 2020年第5期22卷 447-453页

作者： Paula Saez-Espinosa Natalia Huerta-Retamal Laura Robles-Gomez Manuel Aviles Jon Aizpurua Irene Velasco Alejandro Romero Maria Jose Gomez-Torres Department of Biotechnology Faculty of ScienceUniversity of AlicanteAlicante 03080Spain FISABI0-University Hospital of San Juan de Alicante Service of Gynecology and ObstetricsSan Juan de Alicante 03550Spain Departm ent of Cell Biology and Histology Faculty of MedicineUniversity of Murcia and IMIB-ArrixacaMurcia 30100Spain IVF Spain Reproductive MedicineAlicante 03540Spain University Hospital of San Juan de Alicante Service of Gynecology and ObstetricsSan Juan de Alicante 03550Spain Human Fertility Cathedra University of AlicanteAlicante 03080Spain.

A cascade of dramatic physiological events is linked to the sperm acrosome reaction and binding to the oocyte's zona pellucida during human sperm capacitation.However,structural and functional sperm changes during capacitation currently remain poorly defined.Here,we performed a multibiomarker approach based on the utilization of sperm concentration,motility,viability,morphology,acrosome reaction,tyrosine phosphorylation,DNA fragmentation,and lectin-binding sites to analyze the impact caused by swim-up selection times(uncapacitated,1 h capacitated,and 4 h capacitated)on sperm function and structure in normozoospermic samples.We found that a 4 h swim-up capacitation increased sperm quality,because a large number of cells with normal morphology and lower DNA fragmentation rates were recovered.Furthermore,the long-term capacitation induced a higher percentage of cells with tyrosine phosphorylation of the principal piece as well as a redistribution of lectin-binding sites.Overall,the multivariate biomarkers analyzed showed a less variable distribution on spermatozoa recovered after 4 h capacitation than that with the shorter capacitation time.These findings stress the importance of capacitation time as a relevant factor in sperm quality with potential biological reproductive implications both for basic research and in assisted reproduction techniques.

关键词： acrosome reaction capacitation DNA fragmentation Glycocalyx lectin-binding sites membrane integrity protein phosphorylation sperm selection

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Model acupuncture point:Bone marrow-derived stromal stem cells are moved by a weak electromagnetic field

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World Journal of Stem Cells 2016年第10期8卷 342-354页

作者： Artem N Emelyanov Marina V Borisova Vera V Kiryanova Laboratory of High Laser and Magnetic Technology North-Western State Medical University Training Laboratory of Physical Chemistry Institute of ChemistrySt. Petersburg State University Department of Physical Therapy and Rehabilitation North-Western State Medical University

AIM To show the existence of a structural formative role of magnetic fields(MFs) with respect to biological objects by using our proposed model of an acupoint.METHODS We introduced a magnetised 10-100 μT metal rod(needle) into culture dishes with a negatively charged working surface and observed during 24 h how cells were arranged by MFs and by electrical fields(EFs) when attached. Rat and human bone marrow-derived stromal stem cells(r BMSCs and h BMSCs), human nonadherent mononuclear blood cells, NCTCs and A172 cells, and Escherichia coli(E. coli) were evaluated. The dish containing BMSCs was defined as the model of an acupoint. r BMSCs proliferative activity affected by the needle was investigated. For investigating electromagnetic field structures, we used the gas discharge visualisation(GDV) method.RESULTS During 24 h of incubation in 50-mm culture dishes, BMSCs or the nonadherent cells accumulated into a central heap in each dish. BMSCs formed a torus(central ring) with an inner diameter of approximately10 mm only upon the introduction of the needle in the centre of the dish. The cells did not show these effects in 35- or 90-mm culture dishes or hydrophobic dishes or rectangular cuvettes. NCTCs and A172 cells showed unstable the effects and only up to two weeks after thawing. Moreover, we observed that the appearance of these effects depended on the season. In winter, BMSCs showed no the effects. GDV experiments revealed that the resonant annular illumination gradually formed from 10 to 18-20 s in polar solutions with and without cell suspension of BMSCs, NCTCs and E. coli when using circular 50-mm dishes, stimulation at 115 V and switching of the electrode poles at 1 kH z. All these data demonstrate the resonant nature of the central ring. Significant influence of MFs on the rB MSC proliferation rate was not observed.CONCLUSION BMSCs can be moved by MFs when in the presence of a constant EF and MF, when the cells are in the responsive functional state, and when there is a resonant relationship between them.

关键词： Stem cell movement Magnetic targeting Acupuncture Model acupoint Frizzled-related protein Biology resonance Cytoplasm movement Glycocalyx

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Mannosylated glycans impair normal T-cell development by reprogramming commitment and repertoire diversity

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Cellular & Molecular Immunology 2023年第8期20卷 955-968页

作者： Manuel M.Vicente Inês Alves Ângela Fernandes Ana M.Dias Beatriz Santos-Pereira Elena Pérez-Anton Sofia Santos Tao Yang Alexandra Correia Anja Münster-Kühnel Afonso R.M.Almeida Sarina Ravens Gabriel A.Rabinovich Manuel Vilanova Ana E.Sousa SaloméS.Pinho i3S–Institute for Research and Innovation in Health University of PortoPortoPortugal Institute of Biomedical Sciences Abel Salazar(ICBAS) University of PortoPortoPortugal Graduate Program in Areas of Basic and Applied Biology(GABBA) ICBASUniversity of PortoPortoPortugal Faculty of Medicine University of PortoPortoPortugal Nephrology Department Centro Hospitalar e Universitário do PortoPortoPortugal Institute of Immunology Hannover Medical SchoolHannoverGermany Institute of Clinical Biochemistry Hannover Medical SchoolHannoverGermany Instituto de Medicina Molecular João Lobo Antunes Faculdade de MedicinaUniversidade de LisboaLisboaPortugal Laboratorio de Inmunopatología Instituto de Biología y Medicina Experimental(IBYME)Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET)Ciudad de Buenos AiresArgentina Laboratorio de Inmuno-oncología Translacional Instituto de Biología y Medicina Experimental(IBYME)Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET)Ciudad de Buenos AiresArgentina Facultad de Ciencias Exactas y Naturales(FCEyN) Universidad de Buenos AiresCiudad de Buenos AiresArgentina

T-cell development ensures the formation of diverse repertoires of T-cell receptors(TCRs)that recognize a variety of antigens.Glycosylation is a major posttranslational modification present in virtually all cells,including T-lymphocytes,that regulates activity/functions.Although these structures are known to be involved in TCR-selection in DP thymocytes,it is unclear how glycans regulate other thymic development processes and how they influence susceptibility to disease.Here,we discovered stage-specific glycome compositions during T-cell development in human and murine thymocytes,as well as dynamic alterations.After restricting the N-glycosylation profile of thymocytes to high-mannose structures,using specific glycoengineered mice(Rag1CreMgat1fl/fl),we showed remarkable defects in key developmental checkpoints,includingß-selection,regulatory T-cell generation andγδT-cell development,associated with increased susceptibility to colon and kidney inflammation and infection.We further demonstrated that a single N-glycan antenna(modeled in Rag1CreMgat2fl/fl mice)is the sine-qua-non condition to ensure normal development.In conclusion,we revealed that mannosylated thymocytes lead to a dysregulation in T-cell development that is associated with inflammation susceptibility.

关键词： N-glycosylation T-cell development Thymocytes Glycocalyx Inflammation

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HP0953-hypothetical virulence factor overexpresion and localization during Helicobacter pylori infection of gastric epithelium

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World Journal of Gastroenterology 2022年第29期28卷 3886-3902页

作者： Nancy K Arteaga-Resendiz Gerardo E Rodea Rosa María Ribas-Aparicio Alma L Olivares-Cervantes Juan Arturo Castelán-Vega Joséde Jesús Olivares-Trejo Sandra Mendoza-Elizalde Edgar O López-Villegas Christian Colín Pamela Aguilar-Rodea Alfonso Reyes-López Marcela Salazar García Norma Velázquez-Guadarrama Laboratorio de Investigación en Enfermedades Infecciosas Hospital Infantil de México Federico GómezMexico City 06720Mexico Posgrado en Biomedicina y Biotecnología Molecular Laboratorio de Producción y Control de BiológicosEscuela Nacional de Ciencias BiológicasInstituto Politécnico NacionalMexico City 11340Mexico Laboratorio de Adquisición de Hierro Universidad Autónoma de la Ciudad MéxicoPosgrado Ciencias GenómicaMexico City 03100Mexico Laboratorio Central de Microscopía Escuela Nacional de Ciencias BiológicasInstituto Politécnico NacionalMexico City 11340Mexico Centro de estudios económicos y sociales en salud Hospital Infantil de México Federico GómezMexico City 06720Mexico Laboratorio de Investigación en Biología del Desarrollo y Teratogénesis Experimental Hospital Infantil de México Federico GómezMexico City 06720Mexico

BACKGROUND The high prevalence and persistence of Helicobacter pylori(H. pylori) infection, as well as the diversity of pathologies related to it, suggest that the virulence factors used by this microorganism are varied. Moreover, as its proteome contains 340hypothetical proteins, it is important to investigate them to completely understand the mechanisms of its virulence and survival. We have previously reported that the hypothetical protein HP0953 is overexpressed during the first hours of adhesion to inert surfaces, under stress conditions, suggesting its role in the environmental survival of this bacterium and perhaps as a virulence factor.AIM To investigate the expression and localization of HP0953 during adhesion to an inert surface and against gastric(AGS) cells.METHODS Expression analysis was performed for HP0953 during H. pylori adhesion. HP0953 expression at 0,3, 12, 24, and 48 h was evaluated and compared using the Kruskal-Wallis equality-of-populations rank test. Recombinant protein was produced and used to obtain polyclonal antibodies for immunolocalization. Immunogold technique was performed on bacterial sections during adherence to inert surfaces and AGS cells, which was analyzed by transmission electron microscopy. HP0953 protein sequence was analyzed to predict the presence of a signal peptide and transmembrane helices, both provided by the ExPASy platform, and using the GLYCOPP platform for glycosylation sites. Different programs, via, I-TASSER, RaptorX, and HHalign-Kbest, were used to perform three-dimensional modeling.RESULTS HP0953 exhibited its maximum expression at 12 h of infection in gastric epithelium cells.Immunogold technique revealed HP0953 localization in the cytoplasm and accumulation in some peripheral areas of the bacterial body, with greater expression when it is close to AGS cells.Bioinformatics analysis revealed the presence of a signal peptide that interacts with the transmembrane region and then allows the release of the protein to the external environment. The programs also

关键词： Hypothetical protein HP0953 Adherence Helicobacter pylori Glycocalyx Virulence factor Persistence

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