Effects of Fumonisin B1 on Biomechanics and Cytoskeleton of Human Umbilical Vein Endothelial Cells

Objective Fumonisin B1(FB1)is an important mycotoxin in nature worldwide.The biomechanical properties of cells are closely related to their structure and function,and the cytoskeleton is the structural and functional basis of cells motility,and therefore,from a biomechanical point of view,the purpose of this study is to investigate the effects of FB1 on the biomechanical properties,migration capacity and cytoskeletal structure of human umbilical vein endothelial cells(HUVECs),which may lay an experimental foundation for further exploration of the toxicity mechanism of fumonisin.Methods HUVECs were cultured and treated with different concentrations of FB1.Then,CCK-8 kit was used to detect the effect of FB1 on the survival rate.The osmotic fragility of the cells was measured after treatment with different osmotic pressures for30 min.The cell membrane fluidity was measured by fluorescence polarization method.The cell electrophoretic mobility was measured by cell electrophoretic apparatus.The migration capacity of the cells was observed by scratch repair assay.The changes of reactive oxygen species and cytoskeletal structure were observed by confocal laser scanning microscopy.Finally,the mRNA and protein relative expression levels of cytoskeletal binding proteins were detected by real-time PCR,Western blotting and confocal laser scanning.Results The results of CCK-8 showed that FB1 could significantly inhibit the proliferation of HUVECs in a dose-and time-dependent manner.After treatment of HUVECs with FB1,the hypotonic resistance of the cell,cell surface charge,cell membrane fluidity and migration capacity were all weakened,while reactive oxygen species were significantly increased and the cytoskeletal structure was significantly reorganized.Furthermore,RTPCR results showed that the mRNA relative expression levels of cytoskeletal binding proteins,exception of actin,were down-regulated after treated with FB1.Besides,Western blotting and statistical analysis based on fluorescence intensity of laser confocal microscopy confirmed theses changes in protein level.Conclusions FB1 can significantly affect the biomechanical properties and motility of HUVECs,which may be directly correlated to the remodel of F-actin cytoskeleton,as well as the relative expression changes of cytoskeletal binding proteins.It is significant for further exploring the toxicity mechanism of fumonisin.

Effects of Interleukin-10 on the Protein Expression of Human Mature Dendritic Cells

Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.In view of their exceptional ability to present antigen and to interact with T cells,DC play distinct roles in linking innate and adaptive immune responses and thus become logical targets for cancer immunotherapy.Evidences show that tumor-derived cytokines could impair DCs’biomechanics properties,which lead to inefficacy of DCs-based immune therapies.Our previous studies found that IL-10,as one of the widespread suppressive cytokines from tumor microenvironment(TME),could deteriorate DCs’motility and biomechanics properties while the underlying mechanism is unknown.In this study,CD14+monocytes were induced to differentiate into mature dendritic cells after isolation in vitro using recombinant cytokines IL-4,GM-CSF,LPS,and IFN-γ.And we also have compared the proteomic changes of mDCs treated by IL-10 and control group via two-dimensional electrophoresis combine with MALDI-TOF/TOF MS.Then we analyzed the function of differentially expressed proteins through bioinformatics methods include GO analysis that clarified the biological functions of differential proteins and KEGG analysis which enriched signal pathways of differential proteins to explore the molecular mechanism of IL-10 which has inhibitory effect on mDCs.The results showed that IL-10 significantly affected the morphology of mDCs,especially reducing the number and length of filopodia.Different expressed proteins were analyzed by two-dimensional electrophoresis combined with bioinformatics analysis to enrich for glycolytic signaling pathway,HIF-1 signaling pathway and cytoskeletal binding protein expression changes.The results of two-dimensional electrophoresis were verified by Western blot,and the results showed that the data were reliable.In addition,the intracellular ROS levels were significantly higher in mDCs treated with IL-10,validating the previously enriched HIF-1 signaling pathway.In summary,it indicated that IL-10 may interfered with the oxidative metabolic process,glycolytic metabolism,and expression of cytoskeleton-related proteins in mDCs,and disturbances in these physiological processes resulted in reduced biomechanics properties and motility of mDCs and subsequently impaired their immune functions,making DC-based tumor vaccines less effective than which we desired.Our study reveals alterations in the physiological metabolism of mDCs under IL-10 treatment from the proteome,which lays the foundation for further exploration of the altered state of mDCs in the tumor microenvironment.

Sequence-Based Predicting Bacterial Essential ncRNAs Algorithm by Machine Learning

Essential ncRNA is a type of ncRNAwhich is indispensable for the sur-vival of organisms.Although essential ncRNAs cannot encode proteins,they are as important as essential coding genes in biology.They have got wide variety of applications such as antimicrobial target discovery,minimal genome construction and evolution analysis.At present,the number of species required for the deter-mination of essential ncRNAs in the whole genome scale is still very few due to the traditional methods are time-consuming,laborious and costly.In addition,tra-ditional experimental methods are limited by the organisms as less than 1%of bacteria can be cultured in the laboratory.Therefore,it is important and necessary to develop theories and methods for the recognition of essential non-coding RNA.In this paper,we present a novel method for predicting essential ncRNA by using both compositional and derivative features calculated by information theory of ncRNA sequences.The method was developed with Support Vector Machine(SVM).The accuracy of the method was evaluated through cross-species cross-vali-dation and found to be between 0.69 and 0.81.It shows that the features we selected have good performance for the prediction of essential ncRNA using SVM.Thus,the method can be applied for discovering essential ncRNAs in bacteria.

Triboelectric nanogenerators for clinical diagnosis and therapy:A report of recent progress

Triboelectric nanogenerators(TENGs)are considered as an ideal platform for power harvesting for living organisms,thanks to their unique characteristics like flexibility,conversion efficient,and manufacturing cost.Recent advances in TENGs have brought innovative solutions for clinical healthcare.Particularly,TENGs offer novel solutions of continues power supply for wearable and implantable medical devices with lightweight,thinness,good biocompatibility,and excellent soft tissue conformability.In this review,we discuss(1)The working principle and representative structure of TENGs,(2)the material selection of TENGs,(3)the recent progression of application of TENG in the medical field of cardiovascular system,nervous system,respiratory system,microbial inactivation,antibiofouling,disinfection,and tissue repair,(4)challenges and future perspectives of TENG-based medical devices.The emerging TENGs and their applications in medicine cannot simply be seen as an alternative to conventional power supplies,it provides a revolutionary solution for wearable and implantable medical devices,and they will surely change the paradigm of disease diagnosis and treatment in the future.