基于集成技术调控的烟草青枯病绿色防控技术应用研究——以遵义市桐梓县九坝镇白盐井村为例

青枯病是烟叶生产中最难防控、导致损失最大的根茎病害之一.为集成可持续的防控技术体系,本研究针对遵义市桐梓县烟草早生快发慢、青枯病危害严重的实际问题,在该地区连续3年的实践基础上,集成了土壤调酸、根际微生态调控、营养平衡和烟株抗性提升技术的本地化绿色防控技术体系,并评价了集成技术对烟草青枯病的控制效果,及对烟草农艺性状和产质量的影响.结果显示,处理区土壤理化性质得以改善,集成技术对处理区烤烟的生长具有一定的促生作用,全育期增加7 d,对团棵期和打顶期烟苗促生作用最明显.其中,打顶期株高、茎围、叶片数、最大叶长、最大叶宽和叶面积均显著高于对照区,分别提高了14.33%,42.88%,34.28%,17.60%,19.30%和40.27%.此外,采用集成技术的处理区一定程度上延缓了烟草青枯病的发生,病情指数和发病率均显著低于对照区,发病初期相对防效高达94.87%;发病暴发期的相对防效高达78.91%,发病平稳期的相对防效仍高达71.39%.经济性状方面,相比于对照区,处理区每667 m^(2)增收了489.52元,且烟叶均价提高了1.12元/kg.本研究结果可为烟草青枯病绿色防控技术的研发提供参考.

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.

The Dendritic Cells’ Immunological Behaviors Modulated by the Spatial Confinements of Deposited Fibrin Matrix

Implant materials,as foreign objects to host,can cause various degrees of inflammation in most cases.The inflammation is triggered by a series of immune responses and directly impacts the tissue regeneration process,which determines the outcome of tissue repair.The immune responses are complex process involving numerous immune cells and can be divide into innate immune and adaptive immune responses.Once materials are implanted,innate immune responses are activated under the mediation of several immune cells(e.g.neutrophils and macrophages),meanwhile immature dendritic cells(imDCs)are recruited to the implant sites to recognize,internalize and process antigens.Upon antigen uptake,imDCs gradually differentiate into mature dendritic cells(mDCs)and migrate to secondary lymph nodes.In the lymph nodes,mDCs present processed antigen peptides to naive T lymphocytes and activate their antigen specific proliferation,resulting in initiation of adaptive immune responses.Due to their key position in the immune system,serving to bridge innate and adaptive immunity,DCs are crucial to guiding and modulating the immune responses caused by implanted materials.Therefore,figuring out the response of DCs to implanted materials and the exact role of DCs in tissue healing processes will provide deeper insight for the rational design of biomaterials.Previous studies on the effects of implants on immune functions of DCs are mainly focused on physical and chemical properties of the materials(e.g.released chemical composition,surface chemistry,substrate stiffness and surface topography).All these factors will change the microenvironment of the tissue around implant materials,which affect the immune functions of DCs.However,the change of microenvironment not only directly derives from the physical and chemical properties of the material(intrinsic),but also indirectly results from the remodeled extracellular matrix(ECM)caused by implanted materials.When blood or tissue fluid contact with materials after implantation,proteins(e.g.fibrin and collagen)will absorb and deposit on the surface of implants,leading to a provisionally stable matrix with microporous fibrous-liked network structure.It means that the remodeled ECM can provide adhesion sites for recruited DCs and form spatial confinement.DCs,as a kind of cells that are extremely sensitive to mechanical stimuli,theoretically,can response to the mechanical stimuli coming from spatial confinement of remodeled ECM,which may lead to a series of modulations in their cell morphologies and immune functions.Then,the remodeled ECM is a non-negligible mechanical cue.However,to the best of our knowledge,there is a lack of a simple and effective model to establish the relationship between the immune functions of DCs and remodeled ECM.Most studies on the responses of DCs to implanted materials are still based on suspension culture model,which is the normal status of DCs in vitro culture systems.In addition,the processes by which DC exerts immune functions(both endocytosis and antigen presentation)are dynamically physical interaction.It means that the changes of DCs’immune functions are highly correlated with the changes of their biomechanical characteristics caused by remodeled ECM.In this work,we have found that the ECM was remodeled by a large amount of fibrin matrix deposited on the surface of implants in the early stage of the inflammations following implantation.Thus,we used non-toxic salmon fibrin hydrogels with microporous fibrous-liked network structure to mimic the deposited fibrin matrix.Then,human monocyte-derived DCs were cultured on the surface and inside of the fibrin hydrogels to mimic the different spatial confinement states of fibrin matrix.Our results indicated that cell morphologies and cytoskeleton structures of DCs were regulated by the spatial confinement of fibrin hydrogels,resulting in generating mechanical stimuli for DCs.Furthermore,we have found that the biomechanical characteristics and the immune functions of both imDCs and mDC were also modulated.Considering the changes in surface markers,secreted cytokines and biomechanical characteristics of DCs,it indicates that the tendency and magnitude of modulations were highly associated with the spatial confinement of fibrin hydrogels.This model demonstrated that mechanical stimuli deriving from spatial confinement of deposited fibrin matrix is an important factor for regulating the biomechanical characteristics and immune functions of DCs.

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.

Interleukin-10 concentration-dependently impairs the biophysical characteristics and motility of mature dendritic cells

Tumor microenvironment is composed of the tumor cells,stromal cells,microvascular tissue fluid,constitute small amount of infiltrating cells and cytokines.In recent years,more and more evidence that tumor microenvironment play an important role in tumorigenesis.Tumor cells,immune cells and other mesenchymal cells interact and create an immunosuppressive microenvironment through a variety of immunosuppressive factors which vascular endothelial growth factor（VEGF）,transformed growth factor-β1（TGF-β1）and interleukin-10（IL-10）),which suppress immunology functions and promote tumor cells to escape immune surveillance,ultimately leading to tumor growth and metastasis.Dendritic cells（DCs）,the most potent antigen presenting cell as now known,play a key role in the anti-tumor immune process.The pre-

Tmod1 Affects the Immune Functions and Biophysical Properties of Dendritic Cells Through TLR4 Signaling Pathway and Cytoskeleton Remodeling

Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.

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.

Type A aortic dissection following heart transplantation:A case report

BACKGROUND Cardiac transplantation is considered the standard treatment for refractory endstage heart failure.Worldwide,5074 heart transplantations were performed in 2015.About 100 heart transplants are performed at the authors’center each year.The usual complications of heart transplantation include graft rejection,infection,and graft dysfunction.Aortic dissection after heart transplantation is very rare and is a serious complication that requires a hybrid procedure.CASE SUMMARY A 58-year-old female patient was admitted to Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology in July 2020 because of unprovoked low back pain without precipitating causes.Magnetic resonance imaging and computed tomography angiography showed type A aortic dissection with an aberrant right subclavian artery.After admission,urapidil was used to control blood pressure.Ten days later,the patient underwent ascending aortic and aortic arch replacement,subclavian artery reconstruction,and endovascular repair of abdominal and thoracic aortic aneurysms.A cardiopulmonary bypass was established through the right femoral artery and femoral vein.The aberrant right subclavian artery,innominate artery,left common carotid artery,and left subclavian artery were blocked,and the left and right common carotid arteries were cannulated for bilateral cerebral perfusion.CONCLUSION The right axillary artery could not be selected for cardiopulmonary bypass intubation because of aberrant right subclavian artery.

China,Japan and South Korea:Trade Integration under Economic Globalization

Economic globaliza- tion and regional economic integra- tion are the two ma- jor trends of world economic development.In the practice of regional economic integra- tion,the EU and NAFTA as two successful models,has had a significant impact on world economic pattern.Until July 2007,the global effective free trade agreement (FTA) reached 143.In recent years, FTA among developed and developing countries and regions are on the rise.Within regions,such as ASEAN and Japan,China,South Korea, India,Australia,New Zea- land,there have been 14 FTA (including early harvest plan) in operation.To achieve regional economic coopera- tion in East Asia,the key is to build economic integra- tion among China,Japan and South Korea.Therefore,eco- nomic integration among the three is their inevitable choice in the backdrop of economic globalization.While trade integration is the foundation and prerequisite of economic integration,and the former can promote the formation and development of the latter.

Effects of soybean raffinose on growth performance,digestibility,humoral immunity and intestinal morphology of growing pigs

There are appreciable does of raffinose in soybean,but the impacts of raffinose on pigs are poorly investigated.We used 2 experiments to investigate the influence of soybean raffinose on growth per-formance,digestibility,humoral immunity and intestinal morphology of growing pigs.In Exp.1,a total of 30 crossbred(Duroc×Landrace×Yorkshire)barrows(21.93±0.43 kg)were randomly divided into 3 groups,and were fed with the control diet,the control diets supplemented with 0.2%and 0.5%raffinose,respectively,for 21 d.Results showed that the addition of 0.2%or 0.5%raffinose reduced(P<0.05)average daily feed intake(ADFI),average daily gain(ADG)and nutrient digestibility,and dietary 0.5%raffinose increased the ratio of feed to gain(P<0.05).For serum indexes,dietary 0.5%raffinose decreased growth hormone and increased glucagon-like peptide-2,immunoglobulin G,tumor necrosis factor-α(TNF-α)and interleukin-6 concentration(P<0.05).In Exp.2,a total of 24 crossbred barrows(38.41±0.45 kg)were randomly divided into 3 groups,and were fed with the control diet(ad libitum),the raffinose diet(0.5%raffinose,ad libitum),and the control diet in the same amount as the raffinose group(feed-pair group)for 14 d,respectively.Compared with the control diet,dietary 0.5%raffinose decreased ADFI(P<0.05).Intriguingly,the raffinose group had lower ADG than the feed-pair group,lower nutrient digestibility,lower amylase activity in duodenum,lower amylase,lipase and trypsin ac-tivities in jejunum and higher TNF-αconcentration in serum compared with the other 2 groups,and a higher ratio of villus height to crypt depth compared with the control group(P<0.05).These results showed that soybean raffinose could reduce feed voluntary intake and body gain while improving in-testinal morphology without a significant negative influence on immunity.Taken together,dietary raffinose could decrease growth performance by reducing both feed intake and nutrient digestibility while inducing humoral immune response of growing pigs.

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.