An Extended Kalman Filter（EKF） is commonly used to fuse raw Global Navigation Satellite System（GNSS） measurements and Inertial Navigation System（INS） derived measurements. However, the Conventional EKF（CEKF） s...An Extended Kalman Filter（EKF） is commonly used to fuse raw Global Navigation Satellite System（GNSS） measurements and Inertial Navigation System（INS） derived measurements. However, the Conventional EKF（CEKF） suffers the problem for which the uncertainty of the statistical properties to dynamic and measurement models will degrade the performance.In this research, an Adaptive Interacting Multiple Model（AIMM） filter is developed to enhance performance. The soft-switching property of Interacting Multiple Model（IMM） algorithm allows the adaptation between two levels of process noise, namely lower and upper bounds of the process noise. In particular, the Sage adaptive filtering is applied to adapt the measurement covariance on line. In addition, a classified measurement update strategy is utilized, which updates the pseudorange and Doppler observations sequentially. A field experiment was conducted to validate the proposed algorithm, the pseudorange and Doppler observations from Global Positioning System（GPS） and Bei Dou Navigation Satellite System（BDS） were post-processed in differential mode.The results indicate that decimeter-level positioning accuracy is achievable with AIMM for GPS/INS and GPS/BDS/INS configurations, and the position accuracy is improved by 35.8%, 34.3% and 33.9% for north, east and height components, respectively, compared to the CEKF counterpartfor GPS/BDS/INS. Degraded performance for BDS/INS is obtained due to the lower precision of BDS pseudorange observations.展开更多
In airborne tracking,the blind Doppler makes the target undetectable,resulting in tracking difficulties. In this paper,we studied most possible blind-Doppler cases and summed them up into two types:targets' intent...In airborne tracking,the blind Doppler makes the target undetectable,resulting in tracking difficulties. In this paper,we studied most possible blind-Doppler cases and summed them up into two types:targets' intentional tangential flying to radar and unintentional flying with large tangential speed. We proposed an interacting multiple model(IMM) particle filter which combines a constant velocity model and an acceleration model to handle maneuvering motions. We compared the IMM particle filter with a previous particle filter solution. Simulation results showed that the IMM particle filter outperforms the method in previous works in terms of tracking accuracy and continuity.展开更多
In order to track ground moving target, a variable structure interacting multiple model （VS-IMM） using mean shift unscented particle filter （MS-UPF） is proposed in this paper. In model-conditioned filtering, sampl...In order to track ground moving target, a variable structure interacting multiple model （VS-IMM） using mean shift unscented particle filter （MS-UPF） is proposed in this paper. In model-conditioned filtering, sample particles obtained from the unscented particle filter are moved towards the maximal posterior density estimation of the target state through mean shift. On the basis of stop model in VS-IMM, hide model is proposed. Once the target is obscured by terrain, the prediction at prior time is used instead of the measurement at posterior time; in addition, the road model set used is not changed. A ground moving target indication （GMTI） radar is employed in three common simulation scenarios of ground target： entering or leaving a road, crossing a junction and no measurement. Two evaluation indexes, root mean square error （RMSE） and average normalized estimation error squared （ANEES）, are used. The results indicate that when the road on which the target moving changes, the tracking accuracy is effectively improved in the proposed algorithm. Moreover, track interruption could be avoided if the target is moving too slowly or masked by terrain.展开更多
AIM:Transforming growth interacting factor (TGIF) is an inhibitor of both transforming growth factor β (TGF-β) and retinoid signaling pathways. Moreover, the activation of MAPK pathway can prolong its half-life. How...AIM:Transforming growth interacting factor (TGIF) is an inhibitor of both transforming growth factor β (TGF-β) and retinoid signaling pathways. Moreover, the activation of MAPK pathway can prolong its half-life. However, its role in carcinogenesis is still unknown. Thus we attempted to investigate the effect of TGIF on biologic behaviors of gastric carcinoma cells.METHODS: Gastric carcinoma cell line, SGC-7901, was stably transfected with plasmid PcDNA3.1-TGIF. Western blotting and cell immunohistochemistry screening for the highly expressing clone of TGIF were employed. The growth of transfected cells was investigated by MTT and colonyformation assays, and apoptosis was measured by flow cytometry (FCM) and transmission electron microscopy.Tumorigenicity of the transfectant cells was also analyzed.RESULTS: TGIF had no effect on the proliferation, cell cycle and apoptosis of SGC-7901 cells, but cellular organelles of cells transfected with TGIF were richer than those of vector control or parental cells. Its clones were smaller than the control ones in plate efficiency, and its tumor tissues also had no obvious necrosis compared with the vector control or parental cells. Moreover, TGIF could resist TGF-β mediated growth inhibition.CONCLUSION: TGIF may induce differentiation of stomach neoplastic cells. In addition, TGIF can counteract the growth inhibition induced by TGF-β.展开更多
Mitophagy is one of the important targets for the prevention and treatment of myocardial ischemia/reperfusion injury(MIRI).Moderate mitophagy can remove damaged mitochondria,inhibit excessive reactive oxygen species a...Mitophagy is one of the important targets for the prevention and treatment of myocardial ischemia/reperfusion injury(MIRI).Moderate mitophagy can remove damaged mitochondria,inhibit excessive reactive oxygen species accumulation,and protect mitochondria from damage.However,excessive enhancement of mitophagy greatly reduces adenosine triphosphate production and energy supply for cell survival,and aggravates cell death.How dysfunctional mitochondria are selectively recognized and engulfed is related to the interaction of adaptors on the mitochondrial membrane,which mainly include phosphatase and tensin homolog deleted on chromosome ten(PTEN)-induced kinase 1/Parkin,hypoxia-inducible factor-1α/Bcl-2 and adenovirus e1b19k Da interacting protein 3,FUN-14 domain containing protein 1 receptor-mediated mitophagy pathway and so on.In this review,the authors briefly summarize the main pathways currently studied on mitophagy and the relationship between mitophagy and MIRI,and incorporate and analyze research data on prevention and treatment of MIRI with Chinese medicine,thereby provide relevant theoretical basis and treatment ideas for clinical prevention of MIRI.展开更多
In order to realize the aircraft trajectory prediction,a modified interacting multiple model(M-IMM) algorithm is proposed,which is based on the performance analysis of the standard interacting multiple model(IMM) algo...In order to realize the aircraft trajectory prediction,a modified interacting multiple model(M-IMM) algorithm is proposed,which is based on the performance analysis of the standard interacting multiple model(IMM) algorithm.In the proposed M-IMM algorithm,a new likelihood function is defined for the sake of updating flight mode probabilities,in which the influences of interacting to residual's mean error are taken into account and the assumption of likelihood function being a zero mean Gaussian function is discarded.Finally,the proposed M-IMM algorithm is applied to the simulation of the aircraft trajectory prediction,and the comparative studies are conducted to existing algorithms.The simulation results indicate the proposed M-IMM algorithm can predict aircraft trajectory more quickly and accurately.展开更多
Combining interacting multiple model （IMM） and unscented particle filter （UPF）, a new multiple model filtering algorithm is presented. Multiple models can be adapted to targets＇ high maneu- vering. Particle filte...Combining interacting multiple model （IMM） and unscented particle filter （UPF）, a new multiple model filtering algorithm is presented. Multiple models can be adapted to targets＇ high maneu- vering. Particle filter can be used to deal with the nonlinear or non-Gaussian problems and the unscented Kalman filter （UKF） can improve the approximate accuracy. Compared with other interacting multiple model algorithms in the simulations, the results demonstrate the validity of the new filtering method.展开更多
Background: Necroptosis plays an important role in human atherosclerosis and atheroma development. Since receptor interacting protein kinase-3 (RIP3) acts as a key mediator of necroptosis, this study aimed to explore ...Background: Necroptosis plays an important role in human atherosclerosis and atheroma development. Since receptor interacting protein kinase-3 (RIP3) acts as a key mediator of necroptosis, this study aimed to explore its relationship between plasma RIP3 levels and coronary artery disease (CAD) and discover a potential new biomarker for screening CAD subtypes and severity. Methods: A total of 318 patients with CAD who had coronary angiography and 166 controls in Peking Union Medical College Hospital from September 2017 to January 2018 were enrolled in this study. Patients with CAD were divided into three subgroups: patients with stable coronary artery disease (SCAD), patients with unstable angina (UA), and patients with myocardial infarction (MI). The severity of atherosclerosis was determined by Gensini score (GSS). Logistic regression was used to determine the relationship between plasma RIP3 levels and CAD. The correlation between plasma RIP3 and GSS was calculated using multiple linear regression models. Results: Overall, plasma RIP3 levels were significantly higher than serum RIP3 levels. Plasma RIP3 levels in patients with CAD were significantly higher than those in controls. Plasma RIP3 levels were strongly associated with CAD (odds ratio: 6.00, 95% confidence interval 3.04–11.81;P < 0.001). Plasma RIP3 levels increased linearly from controls to patients with SCAD, then patients with UA, and finally to patients with MI. We found a significantly positive correlation between proportion of cases of acute coronary syndrome in subjects and their plasma RIP3 level quartile. Plasma RIP3 levels were also associated with GSS (B 0.027;standard error 0.012;P < 0.05). Conclusions: Plasma RIP3 levels were independently associated with CAD. Plasma RIP3 levels could potentially supplement clinical assessment to screen CAD and determine CAD severity.展开更多
Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs fr...Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs from plants remains challenging.A variety of methods have been utilized to isolate plant EVs from apoplastic washing fluid(AWF).Here,we compare published plant EV isolation methods,and provide our recommended method for the isolation and purification of plant EVs.This method includes a detailed protocol for clean AWF collection from Arabidopsis thaliana leaves,followed by EV isolation via differential centrifugation.To further separate and purify specific subclasses of EVs from heterogeneous vesicle populations,density gradient ultracentrifugation and immunoaffinity capture are then utilized.We found that immunoaffinity capture is the most precise method for specific EV subclass isolation when suitable specific EV biomarkers and their corresponding antibodies are available.Overall,this study provides a guide for the selection and optimization of EV isolation methods for desired downstream applications.展开更多
BACKGROUND Receptor interacting protein kinase 1(RIPK1)-mediated cell death,including apoptosis and necroptosis,belongs to programmed cell death.It has been reported that RIPK1-mediated necroptosis exists in lesions o...BACKGROUND Receptor interacting protein kinase 1(RIPK1)-mediated cell death,including apoptosis and necroptosis,belongs to programmed cell death.It has been reported that RIPK1-mediated necroptosis exists in lesions of cerebral hemorrhage(CH).Electroacupuncture,a treatment derived from traditional Chinese medicine,could improve neurological impairment in patients with brain injury.AIM To investigate the protective role of cross electro-nape acupuncture(CENA)in CH,and clarify the potential mechanism.METHODS CH rat models were established,and CENA was applied to the experimental rats.Neurological functions and encephaledema were then measured.Necrotic cells in the brain of rats with CH were evaluated by propidium iodide staining.Necroptosis was assessed by immunofluorescence.Activation of the necroptosisrelated pathway was detected by western blot.Extraction of brain tissue,cerebrospinal fluid and serum samples was conducted to measure the expression and secretion of inflammatory cytokines by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay.RESULTS The necroptotic marker p-MLKL was detectable in the brains of rats with CH.Next,we found that CENA could ameliorate neurological functions in rat models of CH.Moreover,the upregulation of RIPK1-mediated necroptosis-related molecules in the brains of rats with CH were inhibited by CENA.Further investigation revealed that CENA partially blocked the interaction between RIPK1 and RIPK3.Finally,in vivo assays showed that CENA decreased the expression of the inflammatory cytokines tumor necrosis factor-α,interleukin-6 and interleukin-8 in CH rat models.CONCLUSION These findings revealed that CENA exerts a protective role in CH models by inhibiting RIPK1-mediated necroptosis.展开更多
Tissue and systemic inflammation have been the main culprit behind the cellular response to multiple insults and maintaining homeostasis.Obesity is an independent disease state that has been reported as a common risk ...Tissue and systemic inflammation have been the main culprit behind the cellular response to multiple insults and maintaining homeostasis.Obesity is an independent disease state that has been reported as a common risk factor for multiple metabolic and microvascular diseases including nonalcoholic fatty liver disease(NAFLD),retinopathy,critical limb ischemia,and impaired angiogenesis.Sterile inflammation driven by high-fat diet,increased formation of reactive oxygen species,alteration of intracellular calcium level and associated release of inflammatory mediators,are the main common underlying forces in the pathophysiology of NAFLD,ischemic retinopathy,stroke,and aging brain.This work aims to examine the contribution of the pro-oxidative and pro-inflammatory thioredoxin interacting protein(TXNIP)to the expression and activation of NLRP3-inflammasome resulting in initiation or exacerbation of sterile inflammation in these disease states.Finally,the potential for TXNIP as a therapeutic target and whether TXNIP expression can be modulated using natural antioxidants or repurposing other drugs will be discussed.展开更多
AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice w...AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice werefed normal chow and high fat(HF) diets for 12 wk. The body weight was assessed once weekly. After 12 wk, the liver and serum samples were extracted. The liver tissue expression levels of RIP3, microsomal triglyceride transfer protein, protein disulfide isomerase, apolipoprotein-B, X-box binding protein-1, sterol regulatory element-binding protein-1c, fatty acid synthase, cluster of differentiation-36, diglyceride acyltransferase, peroxisome proliferator-activated receptor alpha, tumor necrosis factor-alpha(TNF-α), and interleukin-6 were assessed. Oleic acid treated primary hepatocytes from WT and RIP3 KO mice were stained with Nile red. The expression of inflammatory cytokines, including chemokine(C-X-C motif) ligand(CXCL) 1, CXCL2, and TNF-α, in monocytes was evaluated.RESULTS RIP3 KO HF diet fed mice showed a significant gain in body weight, and liver weight, liver to body weight ratio, and liver triglycerides were increased in HF diet fed RIP3 KO mice compared to HF diet fed WT mice. RIP3 KO primary hepatocytes also had increased intracellular fat droplets compared to WT primary hepatocytes after oleic acid treatment. RIP3 overexpression decreased hepatic fat content. Quantitative real-time polymerase chain reaction analysis showed that the expression of very-low-density lipoproteins secretion markers(microsomal triglyceride transfer protein, protein disulfide isomerase, and apolipoprotein-B) was significantly suppressed in RIP3 KO mice. The overall NAFLD Activity Score was the same between WT and RIP3 KO mice; however, RIP3 KO mice had increased fatty change and decreased lobular inflammation compared to WT mice. Inflammatory signals(CXCL1/2, TNF-α, and interleukin-6) increased after lipopolysaccharide and pancaspase inhibitor(necroptotic condition) treatment in monocytes. Neutrophil chemok展开更多
基金co-supported by the National Key Research and Development Program of China(No.2016YFC0803103)Beijing Advanced Innovation Center for Future Urban Design(No.UDC2016050100)Beijing Postdoctoral Research Foundation
文摘An Extended Kalman Filter（EKF） is commonly used to fuse raw Global Navigation Satellite System（GNSS） measurements and Inertial Navigation System（INS） derived measurements. However, the Conventional EKF（CEKF） suffers the problem for which the uncertainty of the statistical properties to dynamic and measurement models will degrade the performance.In this research, an Adaptive Interacting Multiple Model（AIMM） filter is developed to enhance performance. The soft-switching property of Interacting Multiple Model（IMM） algorithm allows the adaptation between two levels of process noise, namely lower and upper bounds of the process noise. In particular, the Sage adaptive filtering is applied to adapt the measurement covariance on line. In addition, a classified measurement update strategy is utilized, which updates the pseudorange and Doppler observations sequentially. A field experiment was conducted to validate the proposed algorithm, the pseudorange and Doppler observations from Global Positioning System（GPS） and Bei Dou Navigation Satellite System（BDS） were post-processed in differential mode.The results indicate that decimeter-level positioning accuracy is achievable with AIMM for GPS/INS and GPS/BDS/INS configurations, and the position accuracy is improved by 35.8%, 34.3% and 33.9% for north, east and height components, respectively, compared to the CEKF counterpartfor GPS/BDS/INS. Degraded performance for BDS/INS is obtained due to the lower precision of BDS pseudorange observations.
基金Project supported by China Postdoctoral Science Foundation (No.20060400313)partly by Zhejiang Postdoctoral Science Founda-tion of China (No. 2006-bsh-25)
文摘In airborne tracking,the blind Doppler makes the target undetectable,resulting in tracking difficulties. In this paper,we studied most possible blind-Doppler cases and summed them up into two types:targets' intentional tangential flying to radar and unintentional flying with large tangential speed. We proposed an interacting multiple model(IMM) particle filter which combines a constant velocity model and an acceleration model to handle maneuvering motions. We compared the IMM particle filter with a previous particle filter solution. Simulation results showed that the IMM particle filter outperforms the method in previous works in terms of tracking accuracy and continuity.
文摘In order to track ground moving target, a variable structure interacting multiple model （VS-IMM） using mean shift unscented particle filter （MS-UPF） is proposed in this paper. In model-conditioned filtering, sample particles obtained from the unscented particle filter are moved towards the maximal posterior density estimation of the target state through mean shift. On the basis of stop model in VS-IMM, hide model is proposed. Once the target is obscured by terrain, the prediction at prior time is used instead of the measurement at posterior time; in addition, the road model set used is not changed. A ground moving target indication （GMTI） radar is employed in three common simulation scenarios of ground target： entering or leaving a road, crossing a junction and no measurement. Two evaluation indexes, root mean square error （RMSE） and average normalized estimation error squared （ANEES）, are used. The results indicate that when the road on which the target moving changes, the tracking accuracy is effectively improved in the proposed algorithm. Moreover, track interruption could be avoided if the target is moving too slowly or masked by terrain.
文摘AIM:Transforming growth interacting factor (TGIF) is an inhibitor of both transforming growth factor β (TGF-β) and retinoid signaling pathways. Moreover, the activation of MAPK pathway can prolong its half-life. However, its role in carcinogenesis is still unknown. Thus we attempted to investigate the effect of TGIF on biologic behaviors of gastric carcinoma cells.METHODS: Gastric carcinoma cell line, SGC-7901, was stably transfected with plasmid PcDNA3.1-TGIF. Western blotting and cell immunohistochemistry screening for the highly expressing clone of TGIF were employed. The growth of transfected cells was investigated by MTT and colonyformation assays, and apoptosis was measured by flow cytometry (FCM) and transmission electron microscopy.Tumorigenicity of the transfectant cells was also analyzed.RESULTS: TGIF had no effect on the proliferation, cell cycle and apoptosis of SGC-7901 cells, but cellular organelles of cells transfected with TGIF were richer than those of vector control or parental cells. Its clones were smaller than the control ones in plate efficiency, and its tumor tissues also had no obvious necrosis compared with the vector control or parental cells. Moreover, TGIF could resist TGF-β mediated growth inhibition.CONCLUSION: TGIF may induce differentiation of stomach neoplastic cells. In addition, TGIF can counteract the growth inhibition induced by TGF-β.
基金Supported by the Beijing University of Chinese Medicine Research and Innovation Team Project(No.2019-JYB-TD-08)Youth Science Fund Project of National Natural Science Foundation of China(No.81803906)。
文摘Mitophagy is one of the important targets for the prevention and treatment of myocardial ischemia/reperfusion injury(MIRI).Moderate mitophagy can remove damaged mitochondria,inhibit excessive reactive oxygen species accumulation,and protect mitochondria from damage.However,excessive enhancement of mitophagy greatly reduces adenosine triphosphate production and energy supply for cell survival,and aggravates cell death.How dysfunctional mitochondria are selectively recognized and engulfed is related to the interaction of adaptors on the mitochondrial membrane,which mainly include phosphatase and tensin homolog deleted on chromosome ten(PTEN)-induced kinase 1/Parkin,hypoxia-inducible factor-1α/Bcl-2 and adenovirus e1b19k Da interacting protein 3,FUN-14 domain containing protein 1 receptor-mediated mitophagy pathway and so on.In this review,the authors briefly summarize the main pathways currently studied on mitophagy and the relationship between mitophagy and MIRI,and incorporate and analyze research data on prevention and treatment of MIRI with Chinese medicine,thereby provide relevant theoretical basis and treatment ideas for clinical prevention of MIRI.
基金National Natural Science Foundation of China(No.71401072)Natural Science Foundation of Jiangsu Province,China(No.BK20130814)Fundamental Research Funds for the Central Universities,China(No.NS2013064)
文摘In order to realize the aircraft trajectory prediction,a modified interacting multiple model(M-IMM) algorithm is proposed,which is based on the performance analysis of the standard interacting multiple model(IMM) algorithm.In the proposed M-IMM algorithm,a new likelihood function is defined for the sake of updating flight mode probabilities,in which the influences of interacting to residual's mean error are taken into account and the assumption of likelihood function being a zero mean Gaussian function is discarded.Finally,the proposed M-IMM algorithm is applied to the simulation of the aircraft trajectory prediction,and the comparative studies are conducted to existing algorithms.The simulation results indicate the proposed M-IMM algorithm can predict aircraft trajectory more quickly and accurately.
文摘Combining interacting multiple model （IMM） and unscented particle filter （UPF）, a new multiple model filtering algorithm is presented. Multiple models can be adapted to targets＇ high maneu- vering. Particle filter can be used to deal with the nonlinear or non-Gaussian problems and the unscented Kalman filter （UKF） can improve the approximate accuracy. Compared with other interacting multiple model algorithms in the simulations, the results demonstrate the validity of the new filtering method.
基金the National Natural Science Foundation of China (No. 81670329 and No. 81601431)Chinese Academy of Medical Sciences Initiative for Innovative Medicine (No. 2017-I2M-2-002 and No. 2016-I2M-3-011).
文摘Background: Necroptosis plays an important role in human atherosclerosis and atheroma development. Since receptor interacting protein kinase-3 (RIP3) acts as a key mediator of necroptosis, this study aimed to explore its relationship between plasma RIP3 levels and coronary artery disease (CAD) and discover a potential new biomarker for screening CAD subtypes and severity. Methods: A total of 318 patients with CAD who had coronary angiography and 166 controls in Peking Union Medical College Hospital from September 2017 to January 2018 were enrolled in this study. Patients with CAD were divided into three subgroups: patients with stable coronary artery disease (SCAD), patients with unstable angina (UA), and patients with myocardial infarction (MI). The severity of atherosclerosis was determined by Gensini score (GSS). Logistic regression was used to determine the relationship between plasma RIP3 levels and CAD. The correlation between plasma RIP3 and GSS was calculated using multiple linear regression models. Results: Overall, plasma RIP3 levels were significantly higher than serum RIP3 levels. Plasma RIP3 levels in patients with CAD were significantly higher than those in controls. Plasma RIP3 levels were strongly associated with CAD (odds ratio: 6.00, 95% confidence interval 3.04–11.81;P < 0.001). Plasma RIP3 levels increased linearly from controls to patients with SCAD, then patients with UA, and finally to patients with MI. We found a significantly positive correlation between proportion of cases of acute coronary syndrome in subjects and their plasma RIP3 level quartile. Plasma RIP3 levels were also associated with GSS (B 0.027;standard error 0.012;P < 0.05). Conclusions: Plasma RIP3 levels were independently associated with CAD. Plasma RIP3 levels could potentially supplement clinical assessment to screen CAD and determine CAD severity.
基金supported by grants from the National Natural Science Foundation of China(32070288)supported by grants from the National Institute of Health(R35 GM136379)+3 种基金the National Science Foundation(IOS2017314)the United States Department of Agriculture National Institute of Food and Agriculture(2021-67013-34258 and 2019-70016-29067)the Australian Research Council Industrial Transformation Research Hub(IH190100022)the CIFAR Fungal Kingdom fellowship to H.J。
文摘Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs from plants remains challenging.A variety of methods have been utilized to isolate plant EVs from apoplastic washing fluid(AWF).Here,we compare published plant EV isolation methods,and provide our recommended method for the isolation and purification of plant EVs.This method includes a detailed protocol for clean AWF collection from Arabidopsis thaliana leaves,followed by EV isolation via differential centrifugation.To further separate and purify specific subclasses of EVs from heterogeneous vesicle populations,density gradient ultracentrifugation and immunoaffinity capture are then utilized.We found that immunoaffinity capture is the most precise method for specific EV subclass isolation when suitable specific EV biomarkers and their corresponding antibodies are available.Overall,this study provides a guide for the selection and optimization of EV isolation methods for desired downstream applications.
基金Supported by State Administration of Traditional Chinese Medicine of Heilongjiang Province,No.ZHY16-027Harbin Municipal Science and Technology BureauYouth Reserve Talent Project,No.2017RAQXJ170
文摘BACKGROUND Receptor interacting protein kinase 1(RIPK1)-mediated cell death,including apoptosis and necroptosis,belongs to programmed cell death.It has been reported that RIPK1-mediated necroptosis exists in lesions of cerebral hemorrhage(CH).Electroacupuncture,a treatment derived from traditional Chinese medicine,could improve neurological impairment in patients with brain injury.AIM To investigate the protective role of cross electro-nape acupuncture(CENA)in CH,and clarify the potential mechanism.METHODS CH rat models were established,and CENA was applied to the experimental rats.Neurological functions and encephaledema were then measured.Necrotic cells in the brain of rats with CH were evaluated by propidium iodide staining.Necroptosis was assessed by immunofluorescence.Activation of the necroptosisrelated pathway was detected by western blot.Extraction of brain tissue,cerebrospinal fluid and serum samples was conducted to measure the expression and secretion of inflammatory cytokines by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay.RESULTS The necroptotic marker p-MLKL was detectable in the brains of rats with CH.Next,we found that CENA could ameliorate neurological functions in rat models of CH.Moreover,the upregulation of RIPK1-mediated necroptosis-related molecules in the brains of rats with CH were inhibited by CENA.Further investigation revealed that CENA partially blocked the interaction between RIPK1 and RIPK3.Finally,in vivo assays showed that CENA decreased the expression of the inflammatory cytokines tumor necrosis factor-α,interleukin-6 and interleukin-8 in CH rat models.CONCLUSION These findings revealed that CENA exerts a protective role in CH models by inhibiting RIPK1-mediated necroptosis.
文摘Tissue and systemic inflammation have been the main culprit behind the cellular response to multiple insults and maintaining homeostasis.Obesity is an independent disease state that has been reported as a common risk factor for multiple metabolic and microvascular diseases including nonalcoholic fatty liver disease(NAFLD),retinopathy,critical limb ischemia,and impaired angiogenesis.Sterile inflammation driven by high-fat diet,increased formation of reactive oxygen species,alteration of intracellular calcium level and associated release of inflammatory mediators,are the main common underlying forces in the pathophysiology of NAFLD,ischemic retinopathy,stroke,and aging brain.This work aims to examine the contribution of the pro-oxidative and pro-inflammatory thioredoxin interacting protein(TXNIP)to the expression and activation of NLRP3-inflammasome resulting in initiation or exacerbation of sterile inflammation in these disease states.Finally,the potential for TXNIP as a therapeutic target and whether TXNIP expression can be modulated using natural antioxidants or repurposing other drugs will be discussed.
基金Supported by National Research Foundation of Korea(NRF)funded by the South Korean Government,No.NRF-2017M3A9C8028794
文摘AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice werefed normal chow and high fat(HF) diets for 12 wk. The body weight was assessed once weekly. After 12 wk, the liver and serum samples were extracted. The liver tissue expression levels of RIP3, microsomal triglyceride transfer protein, protein disulfide isomerase, apolipoprotein-B, X-box binding protein-1, sterol regulatory element-binding protein-1c, fatty acid synthase, cluster of differentiation-36, diglyceride acyltransferase, peroxisome proliferator-activated receptor alpha, tumor necrosis factor-alpha(TNF-α), and interleukin-6 were assessed. Oleic acid treated primary hepatocytes from WT and RIP3 KO mice were stained with Nile red. The expression of inflammatory cytokines, including chemokine(C-X-C motif) ligand(CXCL) 1, CXCL2, and TNF-α, in monocytes was evaluated.RESULTS RIP3 KO HF diet fed mice showed a significant gain in body weight, and liver weight, liver to body weight ratio, and liver triglycerides were increased in HF diet fed RIP3 KO mice compared to HF diet fed WT mice. RIP3 KO primary hepatocytes also had increased intracellular fat droplets compared to WT primary hepatocytes after oleic acid treatment. RIP3 overexpression decreased hepatic fat content. Quantitative real-time polymerase chain reaction analysis showed that the expression of very-low-density lipoproteins secretion markers(microsomal triglyceride transfer protein, protein disulfide isomerase, and apolipoprotein-B) was significantly suppressed in RIP3 KO mice. The overall NAFLD Activity Score was the same between WT and RIP3 KO mice; however, RIP3 KO mice had increased fatty change and decreased lobular inflammation compared to WT mice. Inflammatory signals(CXCL1/2, TNF-α, and interleukin-6) increased after lipopolysaccharide and pancaspase inhibitor(necroptotic condition) treatment in monocytes. Neutrophil chemok
