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Purpose We analyze the immune infiltration model of osteoarthritis to determine the relevant diagnostic biomarkers (OA), and to provide some help for the treatment and diagnosis of OA. Methods From the Gene Expression Omnibus (GEO) database, we downloaded GSE168505 and GSE114007 gene expression datasets, including 24 patients and 21 healthy controls. The R software Limma package and SVA package were used to analyze the batch effect. We selected differentially expressed genes (DEGs), and we then analyzed the DEGs’ functional enrichment. We performed differential analysis to pick out the differentially expressed immune-related genes (DEIRGs) in the merged data set. We first selected the candidate genes by the least absolute shrinkage and selection operator (LASSO) method, and then further screened the diagnostic markers by support vector machine-recursive feature elimination algorithm (SVM-RFE). In dataset GSE129147, the diagnostic value was determined by drawing the receiver operating characteristic (ROC) curve. In addition, we used the CIBERSORT program to assess the 22 kinds immune cells of infiltration models. Finally, an in vitro cell model of OA was established by interleukin-1β(IL-1β) to verify the bioinformatics results. Results Through differential analysis, 454 differential genes were identified, mainly involved ossification, extracellular matrix organization, collagen − containing extracellular matrix, metalloendopeptidase activity, PI3K − Akt signaling pathway, regulation of cell population proliferation, and other biological processes. We screened BIRC5 and TNFSF11 as candidate biomarkers by machine learning. In the data set GSE129147, BIRC5 and TNFSF11 were verified as diagnostic markers of OA by the ROC curve. The following correlation analysis found that BIRC5 and TNFSF11 were correlated with Mast cells resting, NK cells resting, Monocytes, Plasma cells, Eosinophil, Macrophages M0, and Macrophages M2. The expression of BIRC5 and TNFSF11 was up-regulated in the OA model in vitro. Conclusion We conclude that BIRC5 and TNFSF11 can be biomarkers for diagnosing OA. This discovery provides a direction for the occurrence of OA and the exploration of new treatment methods from the perspective of immunology.

While porous metal-supported solid fuel oxide cells (PMS-SOFCs) have the potential to decrease the cost and increase the start-up speed of power units, the available fabrication processes remain too cumbersome for industrial production. In this study, we prepared bulk-like strontium and magnesium-doped lanthanum gallate (LSGM) coatings using atmospheric plasma spraying (APS) at an increased particle temperature. The large equiaxed grains inside the coatings indicated the epitaxial growth of the splat interfaces and improvement in the coating quality. With increased particle temperature, the conductivity of dense LSGM coatings directly deposited by APS was comparable to that of the bulk material, and cell performance was also significantly enhanced. The maximum power density of the PMS-SOFC at 700 °C was 831 mW cm−2 and 596 mW cm−2 when high and low particle temperatures were used, respectively. These results indicated that the quality of the coating was improved by increasing the in-flight particle temperature.

metal supported solid oxide fuel cells (PMS-SOFCs) are regarded as a promising choice for power generation. 430 L steel is a popular choice for SOFC support due to its acceptable electrical perform...

Wei Cao,* Shuai Song,* Guojian Fang,* Yingze Li, Yuepeng Wang, Qun-Shan Wang Department of Cardiology, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, 200092, People’s Republic of China*These authors contributed equally to this workCorrespondence: Qun-Shan Wang; Yuepeng Wang Email wangqunshan@xinhuamed.com.cn; wangyuepeng@xinhuamed.com.cnBackground: Atrial fibrosis serves as a disease initiating mechanism in the development of atrial fibrillation. Angiotensin II (Ang-II), a key mediator for atrial fibrosis, aberrantly activates atrial fibroblasts (AFs) into myofibroblasts, resulting in subsequent excessive synthesis and deposition of extracellular matrix (ECM). Cadherin-11 (CDH11) is essential in the development of non-cardiac fibrotic diseases. In this study, we investigated its role in the pathogenesis and underlying mechanism of atrial fibrillation.Methods: We obtained left atrial tissues from either patients with atrial fibrillation or Ang-II-induced atrial fibrosis mice. We utilized a global CDH11 knockout mouse (CDH11−/-) model to determine the effect of CDH11 on AF cell proliferation, migration, ECM synthesis/deposition. RNA-Seq of isolated AFs from CDH11−/- or normal mice was performed and differential expressed genes were analyzed. The mouse susceptibility to atrial fibrillation was examined by cardiac electrophysiology.Results: We found that cadherin-11 was significantly up-regulated in fibrotic atrial tissue from patients with atrial fibrillation and Ang-II-induced mice. Both normal and CDH11−/- mice did not develop atrial fibrosis at resting state. However, after Ang-II infusion, unlike severe atrial fibrosis occurred in normal mice, CDH11−/- mice displayed a reduced atrial fibrosis. Atrial fibroblasts with CDH11 deletion from CDH11−/- mice showed reduction in Ang-II-induced cell proliferation, migration and ECM synthesis/deposition, indicating the involvement of CDH11 in atrial fibrosis. Consistently, RNA-Seq of CDH11-null AFs uncovered significant decrease in pro-fibrotic gene expression. In addition, we identified reduction of transcripts associated with Smad2/3, ERK1/2 and JNK pathways. Further, CDH11−/- mice showed a significantly attenuated Ang-II-induced susceptibility to atrial fibrillation.Conclusion: Our results indicate that CDH11 potentiates Ang-II-induced activation of AFs. The pathogenesis of atrial fibrosis is through CDH11 mediated stimulation of Smad2/3, ERK1/2 and JNK pathways. Thus, CDH11 might serve as a novel therapeutic target for ameliorating the development of atrial fibrillation.Keywords: atrial fibrillation, atrial fibrosis, atrial fibroblast, cadherin-11, angiotensin-II

Yuhan Chen,1,* Yuanyuan Chen,1,* Xueze Jiang,1 Mengkun Shi,2 Zhenwei Yang,1 Zhiyong Chen,1 Xuesheng Hua,1 Jie Chen,1 Yuepeng Wang1 1Department of Cardiology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200092, People’s Republic of China; 2Department of Cardio-Thoracic Surgery, Shanghai Tongji Hospital, Tongji University School of Medicine, Shanghai, 200065, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yuepeng Wang; Jie ChenDepartment of Cardiology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200092, People’s Republic of ChinaTel +86 18217267289; +86 15280380968Email wangyuepeng@xinhuamed.com.cn; jieyuepeng@msn.comBackground: Activation of vascular adventitial fibroblasts (VAFs) upon vascular injury contributes greatly to the medial vascular smooth muscle cells (VSMCs) proliferation, migration and the subsequent neointima formation. A number of factors including fibroblast growth factors (FGFs) have been shown to control VSMC growth, proliferation and phenotypic switching, suggesting that they may function as paracrine signals for VAFs to modulate VSMCs functions. However, little is known about the signaling molecule(s) and its mechanism of action. This study is set to identify which and how FGF family members are involved in VAFs mediated vascular remodeling.Methods: We used qPCR, Western blot and Immunohistochemistry to observe the spatiotemporal expression of FGF10 and FGFR2 in injured vascular tissue. The proliferation and migration assays of VSMCs were performed in a co-culture system. The activation of signaling pathway was detected by Western blot, immunohistochemistry and immunofluorescence. Hematoxylin-eosin and immunofluorescence were used to assess the effects of exogenous FGF10 and siFGF10 on the neointima formation.Results: The expression of FGF10 and FGFR2 were increased from day 3 through day 14 post injury. FGF10 was significantly upregulated in adventitia, and FGFR2 was detected in both media and neointima after injury. In vitro, FGF10 was most prominently expressed in VAFs and FGFR2 was significantly expressed in VSMCs. Both were regulated by PDGF. Co-culture of VAFs and VSMCs in vitro showed that VAF-derived FGF10 promoted the proliferation and migration of VSMCs. PDGF could synergistically enhance the process. VAF-derived FGF10 can significantly activate the FGFR2 in VSMCs and furthermore significantly activate the downstream MAPK/PI3K-AKT signaling pathways. Delivery of exogenous FGF10 potentiated the neointima formation, while siFGF10 attenuated the neointima formation.Conclusion: VAFs-derived FGF10 promoted the proliferation and migration of VSMCs and neointima formation, and FGF10-FGFR2 signaling triggered the activation of MAPK/PI3K-AKT pathways in VSMCs and PDGF synergistically amplified FGF10 signaling.Keywords: vascular adventitial fibroblasts, vascular smooth muscle cells, FGF10, proliferation, migration, neointima formation

The modified Pechini method was applied to prepare a highly active and novel cathode material La0.7Sr0.3Ti0.15Fe0.65Ni0.2O3-δ (LSTFN). This material was coated on the LGSM electrolyte through a screen-printing technique with variable thicknesses of 28 ± 8, 41 ± 8, and 62 ± 8 μm, respectively. Different fabrication parameters, including sintering temperature, time, coating thickness, and variations in ball-milling, which affect the electrochemical performance of the cathode material, were investigated. X-ray diffraction analysis of the cathode material suggested that it exhibits a cubic crystal structure with a LSTFN single phase. The morphological studies were conducted using scanning electron microscopy (SEM), which confirmed that the electrode material had a highly porous structure. Meanwhile, the electrochemical properties of the material were studied by electrochemical impedance spectroscopy (EIS), which revealed that by varying different parameters, the electrochemical performance of the electrode material was enhanced. The coated cathode materials with variable thicknesses were analyzed at different sintering temperatures and times. Experimental results suggest that the optimum sintering temperature and time were 950 °C and 3 h, respectively, at which LSTFN exhibits the minimum polarization resistance (RP) of 0.046 Ωcm2 when sintered at 800 °C for 3 h.

This study adopts a novel structure design of large-area planar metal-supported solid oxide fuel cell (MS-SOFC), which exhibits the characteristics of self-sealing and high thermal cycling resistance. The self-sealing structure of MS-SOFCs is achieved by brazing technology. Plasma spraying is performed to prepare all functional layers of cells. The size of cells is 10 × 10 cm2. The coefficients of thermal expansion of relevant parts (i.e., porous metal support, interconnector and compound structure of both) and the electrical conductivity of brazing solder were measured. The microstructure of brazing area was observed in situ at 800 °C for 500 h, and the distribution of fuel gas in the interconnector was simulated. The performance of cells was characterized and found that the maximum power density can reach up to 716 mW cm−2 at 700 °C. Moreover, the long-term stability of the cell was investigated for about 500 h with 6 times of thermal cycling implemented during this period. The open circuit voltage and the maximum power density of the cell showed a good stability.

Atrial fibrillation (AF) is the most common cardiac arrhythmia nowadays. The occurrence of AF is closely associated with obesity. Cadherin-11 (Cad-11), as a member of the cadherin family, can make a contribution to diet-induced obesity and it will be informative to know whether Cad-11 exerts its effects on atrial remodeling and AF vulnerability in a diet-induced obesity model. In this study, we demonstrated that the expression of Cad-11 was significantly upregulated in the left atrium of AF patients with obesity and mice following 16 weeks of high-fat diet (HFD) feeding. Further confirmed that Cad-11 could regulate the activity of atrial fibroblasts by participating in inducing proinflammatory cytokines production. At animal levels, we found that although there was a lack of statistical difference in body weight, Cad-11-/- mice could markedly improve impaired glucose tolerance and hyperlipidemia. Adverse atrial structural remodeling, including atrial enlargement, inflammation, and fibrosis provoked by HFD feeding were mitigated in Cad-11-/- mice. Mechanistically, Cad-11 activated mitogen-activated protein kinases and nuclear factor-κB for interleukin-6 production in atrial fibroblasts that may contribute to the atrial fibrosis process in obesity-related AF, suggesting Cad-11 might be a new therapeutic target for obesity-related AF.

The present meta-analysis was conducted to compare the safety and effectiveness of reverse shoulder arthroplasty (RSA) and hemiarthroplasty (HA) in the treatment of osteoporotic proximal humeral fractures in elderly patients. The Embase, Pubmed Central, Cumulative Index to Nursing and Allied Health Literature, ProQuest Dissertations and Theses, Cochrane Library and Chinese Biomedical databases were searched between January 2009 and January 2022 to identify relevant studies. According to the search strategy, a total of 210 associated studies were retrieved and 16 were finally included. Review Manager 5.4 software was used for the data analysis. This study indicated that patients in the RSA group had significantly improved treatment outcomes compared with patients in the HA group, as assessed by Constant-Murley Shoulder Outcome Score (95% CI, 1.69-3.76; P0.001), American Shoulder and Elbow Surgeons score (95% CI, 11.81-24.88; P0.001) and shoulder range of motion (ROM; 95% CI, 3.41-9.07; P0.001). However, the HA group was superior to the RSA group in terms of the Oxford Shoulder score (95% CI, 2.89-11.11; P0.001). There was no significant statistical difference between the two groups in terms of the Disabilities of the Arm, Shoulder and Hand score and complications. Overall, for the treatment of osteoporotic proximal humeral fractures in the elderly, the RSA group had improved postoperative ROM and functional scores compared with the HA group, without significant difference in the incidence of complications. However, HA remains a safe and reliable treatment option.

A novel self-sealing structure for metal-supported solid oxide fuel cells (MS-SOFCs) is designed by applying brazing technology between the metal support and interconnector to solve the sealing problem on the anode side of planar SOFCs. A high-reliability self-sealing effect is thus realized at the anode side of the MS-SOFC. Plasma spraying technology is used to prepare cell functional layers including the anode, cathode, and electrolyte. A single cell is assembled with a 50–60-μm plasma-sprayed Sc2O3-stabilized ZrO2 (ScSZ) electrolyte layer. The gas permeability of the self-sealed MS-SOFC without a cathode layer is 0.42 × 10−17 m2. The open-circuit voltage of the cell is ~ 1.1 V in the operating temperature range from 550 to 750 °C. The power density of the cell reaches 1109 mW cm−2 at 750 °C under standard atmosphere. In addition, the use of a fuel gas pressure that is 20 kPa higher than the cathodic gas pressure results in a significant increase in the power density to 1782 mW cm−2 at 750 °C. The novel cell structure and gas tightness of the ScSZ electrolyte prepared by plasma spraying indicates that it can meet the requirements of SOFC applications.

A particular YSZ feedstock for very low-pressure plasma spraying (VLPPS) has been designed which can be automatically divided into less than 5 μm YSZ molten particles in the long plasma jet. Fully molten particles were deposited on the substrate at different deposition distances of 250, 350 and 450 mm, respectively. The deposition behavior of less than 5 μm YSZ molten particles were studied aiming to obtain a thin gastight YSZ coating. The flattening ratio of particles at different deposition distances and the gas permeability of YSZ coating prepared by VLPPS were investigated. The results revealed that the thickness of flattened particles was about 0.10-0.35 μm and the flattening ratio of molten particles was about 4.7. The flattened particles were bonded well with substrates and the width of vertical cracks appeared in flattened particles was 0.01-0.02 μm. The gas permeability of coatings prepared at 350 mm was 1.5 × 10−7 cm4 (gf)−1 s−1.

Very low pressure plasma spraying (VLPPS) is an attractive method for metal-supported solid oxide fuel cells, wherein it can significantly avoid the oxidation of metal substrate. In this study, scandia-stabilized zirconia electrolyte was fabricated by VLPPS. To investigate the microstructure of coatings, the spraying distances were set to 150, 250 and 350 mm. The fractured morphology suggests that, at a relatively low deposition temperature (

Angiotensin II (Ang-II)-induced fibroblast differentiation plays an important role in the development of atrial fibrosis and atrial fibrillation (AF). Here, we show that the expression of the histone methyltransferase enhancer of zeste homolog 2 (EZH2) is increased in atrial muscle and atrial fibroblasts in patients with AF, accompanied by significant atrial fibrosis and atrial fibroblast differentiation. In addition, EZH2 is induced in murine models of atrial fibrosis. Furthermore, either pharmacological GSK126 inhibition or molecular silencing of EZH2 can inhibit the differentiation of atrial fibroblasts and the ability to produce ECM induced by Ang-II. Simultaneously, inhibition of EZH2 can block the Ang-II-induced migration of atrial fibroblasts. We found that EZH2 promotes fibroblast differentiation mainly through the Smad signaling pathway and can form a transcription complex with Smad2 to bind to the promoter region of the ACTA2 gene. Finally, our in vivo experiments demonstrated that the EZH2 inhibitor GSK126 significantly inhibited Ang-II-induced atrial enlargement and fibrosis and reduced AF vulnerability. Our results demonstrate that targeting EZH2 or EZH2-regulated genes might present therapeutic potential in AF.

A super long and stable laminar plasma jet was used in the atmospheric plasma spray process in this work. Microstructures evolution and properties of alumina coatings that were obtained using a long spraying distance ranging from 200 mm to 350 mm in an atmospheric environment were studied in this work. These processes were carried out by using a total gas flow rate of 14 slpm with 70% nitrogen and 30% argon in volume and output power of 25.4 kW (current of 160 A). A long particle residence time was obtained in this study, which compared with other current atmospheric plasma spray methods. The microstructures of coatings showed the multi-island protrusions on the top surfaces and overlapped lamellar splats at the fracture surfaces. The microstructural evolution was significantly affected by the heating and motion behaviors of alumina particles when these were flowing in the long laminar plasma jet at different spraying distances.

The multiple scattering of elastic waves by uniformly arranged nanopores and the equivalent properties of nanoporous materials are studied in this paper. By using displacement potential function method, the stress fileds and strain fields around nanopores and the matrix are solved in accordance with the classical elasticity theory and surface elasticity theory. The equivalent bulk moduli and shear moduli of nanoporous materials are studied using the Mori-Tanaka method. Results show that the equivalent properties of nanoporous materials not only depend on the surface parameters but are also affected by the volume percentage of the holes.

Background and purpose 9-Phenanthrol, known as a specific inhibitor of the transient receptor potential melastatin 4 (TRMP4) channel, has been shown to modulate cardiac electrical activity and exert antiarrhythmic effects. However, its pharmacological effects remain to be fully explored. Here, we tested the hypothesis that cardiac sodium current inhibition contributes to the cardioprotective effect of 9-phenanthrol. Experimental approach Single ventricular myocytes (VMs) and Purkinje cells (PCs) were enzymatically isolated from rabbits. Arterially perfused rabbit wedge preparations were also used, and transmural electrocardiogram and endocardial action potentials (APs) were simultaneously recorded. Wild-type and mutated human recombinant SCN5A were expressed in HEK293 cells. Anemonia toxin II (ATX-II) was used to amplify the late sodium current (INaL ) and induce arrhythmias. Whole-cell patch clamp technique was used to record APs and ionic currents. Key results 9-Phenanthrol (10-50 μM) stabilized ventricular repolarization and abolished arrhythmias induced by ATX-II in both isolated VMs, PCs and wedge preparations. Further study revealed that 9-phenanthrol modulated the gating properties of cardiac sodium channels and dose-dependently inhibited INaL and peak sodium current (INaP ) in VMs with an IC50 of 18 and 71.5 μM respectively. Its ability to inhibit INaL was further confirmed in PCs and HEK293 cells expressing SCN5A mutations. Conclusions and implications Our results indicate that 9-phenanthrol modulates the gating properties of cardiac sodium channels and inhibits INaL and INaP , which may contribute to its antiarrhythmic and cardioprotective effects.

Rate-dependent repolarization (RDR) of action potential (AP) in cardiomyocyte plays a critical role in the genesis of arrhythmias and RDR in atrium has been linked with atrial fibrillation. However, detailed studies focusing on the role of RDR in rabbit atrium are scant. In this study, atrial cells were isolated from rabbit heart and rate-dependent property was explored in single atrial cell to elucidate the underlying mechanism. Our results indicated that rate-dependent prolongation was evident at the action potential duration at 20% (APD20) and 50% (APD50) repolarization but not at 90% repolarization (APD90) under control condition. Using transient outward potassium current (Ito) inhibitor 4-Aminopyridine (4-AP, 2 mM) effectively eliminated the changes in APD20 and APD50, and unmasked the rate-dependent reduction of APD90 which could be diminished by further adding L-type calcium current (ICaL) inhibitor nifedipine (30 μM). However, using the selective late sodium current (INaL) inhibitor GS-458967 (GS967, 1 μM) caused minimal effect on APD90 of atrial cells both in the absence and presence of 4-AP. In consistence with results from APs, Ito and ICaL displayed significant rate-dependent reduction because of their slow reactivation kinetics. In addition, the magnitude of INaL in rabbit atrium was so small that its rate-dependent changes were negligible. In conclusion, our study demonstrated that Ito and ICaL mediate RDR of AP in rabbit atrium, while minimal effect of INaL was seen.

Cardiac hypertrophy is characterized by increased myofibrillogenesis. Angiotensin II (Ang-II) is an essential mediator of the pressure overload–induced cardiac hypertrophy in part through RhoA/ROCK (small GTPase/Rho-associated coiled-coil containing protein kinase) pathway. FHOD3 (formin homology 2 domain containing 3), a cardiac-restricted member of diaphanous-related formins, is crucial in regulating myofibrillogenesis in cardiomyocytes. FHOD3 maintains inactive through autoinhibition by an intramolecular interaction between its C- and N-terminal domains. Phosphorylation of the 3 highly conserved residues (1406S, 1412S, and 1416T) within the C terminus (CT) of FHOD3 by ROCK1 is sufficient for its activation. However, it is unclear whether ROCK-mediated FHOD3 activation plays a role in the pathogenesis of Ang-II–induced cardiac hypertrophy. In this study, we detected increases in FHOD3 expression and phosphorylation in cardiomyocytes from Ang-II–induced rat cardiac hypertrophy models. Valsartan attenuated such increases. In cultured neonate rat cardiomyocytes, overexpression of phosphor-mimetic mutant FHOD3-DDD, but not wild-type FHOD3, resulted in myofibrillogenesis and cardiomyocyte hypertrophy. Expression of a phosphor-resistant mutant FHOD3-AAA completely abolished myofibrillogenesis and attenuated Ang-II–induced cardiomyocyte hypertrophy. Pretreatment of neonate rat cardiomyocytes with ROCK inhibitor Y27632 reduced Ang-II–induced FHOD3 activation and upregulation, suggesting the involvement of ROCK activities. Silencing of ROCK2, but not ROCK1, in neonate rat cardiomyocytes, significantly lessened Ang-II–induced cardiomyocyte hypertrophy. ROCK2 can directly phosphorylate FHOD3 at both 1412S and 1416T in vitro and is more potent than ROCK1. Both kinases failed to phosphorylate 1406S. Coexpression of FHOD3 with constitutively active ROCK2 induced more stress fiber formation than that with constitutively active ROCK1. Collectively, our results demonstrated the importance of ROCK2 regulated FHOD3 expression and activation in Ang-II–induced myofibrillogenesis, thus provided a novel mechanism for the pathogenesis of Ang-II–induced cardiac hypertrophy.

Cardiac fibrosis (CF), a repairing process following myocardial infarction (MI), is characterized by abnormal proliferation of cardiac fibroblasts and excessive deposition of extracellular matrix (ECM) resulting in inevitable resultant heart failure. TGF-β (transforming growth factor-β)/ALK5 (Activin receptor-like kinase 5)/Smad2/3/4 pathways have been reported to be involved in the process. Recent studies have implicated both activin and its specific downstream component ALK4 in stimulating fibrosis in non-cardiac organs. We recently reported that ALK4 is upregulated in the pressure-overloaded heart and its partial inhibition attenuated the pressure overload-induced CF and cardiac dysfunction. However, the role of ALK4 in the pathogenesis of MI-induced CF, which is usually more severe than that induced by pressure-overload, remains unknown. Here we report: 1) In a wild-type mouse model of MI, ALK4 upregulation was restricted in the fibroblasts of the infarct border zone; 2) In contrast, ALK4+/- mice with a haplodeficiency of ALK4 gene, showed a significantly attenuated CF in the border zone, with a smaller scar size, a preserved cardiac function and an improved survival rate post-MI; 3) Similarly to pressure-overloaded heart, these beneficial effects might be through a partial inactivation of the Smad3/4 pathway but not MAPK cascades; 4) The apoptotic rate of the cardiomyocytes were indistinguishable in the border zone of the wild-type control and ALK4+/- mice; 5) Cardiac fibroblasts isolated from ALK4+/- mice showed reduced migration, proliferation and ECM synthesis in response to hypoxia. These results indicate that partial inhibition of ALK4 may reduce MI-induced CF, suggesting ALK4 as a novel target for inhibition of unfavorable CF and for preservation of LV systolic function induced by not only pressure-overload but also MI.

This study assesses the potential of scandia-stabilized zirconia (ScSZ) produced by very low-pressure plasma spraying (VLPPS) for metal-supported solid oxide fuel cell (MS-SOFC) applications. To investigate the microstructure of ScSZ, coating samples were deposited at spraying distances of 150, 250, 350 mm. The fragile nature of coating cross-sections suggests that the typical lamellar structure of zirconia is replaced by a transgranular structure. Nonetheless, apparent porosity, ionic conductivity, open circuit voltage, and ohmic resistance measurements indicate that VLPPS is a viable method for producing MS-SOFCs.

Background Activin receptor-like kinase 4 (ALK4), a downstream receptor of transforming growth factor-β superfamily, is highly expressed in the mammal heart. Upregulated ALK4 expression and activated ALK4-small mother against decapentaplegic (Smad)2/3 signaling have been reported to play a pivotal role in tumorigenesis and in the development of systemic sclerosis. However, the role of ALK4-Smad2/3 pathway in the pathogenesis of cardiac hypertrophy and cardiac fibrosis remains unknown. Methods and results In this study, the mice with heterozygous knocking out of ALK4 gene (ALK4) were generated and subjected to aortic banding for 4 weeks. We found that ALK4 expression was upregulated in aortic banding-induced model of cardiac hypertrophy and cardiac fibrosis in wild-type mice. Compared with the wild-type mice, ALK4mice demonstrated a similar extent of aortic banding-induced cardiac hypertrophy, but a significant suppression of cardiac fibrosis to 64.8% of the basal level, and a subsequent amelioration in the cardiac dysfunction (left ventricle ejection fraction: 59.0 ± 6.4 in wild-type mice vs. 75.6 ± 3.9% in ALK4 mice; left ventricle end-diastolic pressure: 16.6 ± 4.7 mmHg in wild-type mice vs. 6.6 ± 2.8 mmHg in ALK4 mice) associated with inhibition of cardiac fibroblast activation and cardiomyocyte apoptosis. In vitro, ALK4 haploinsufficiency blocked the cellular proliferation/differentiation and collagen production in cultured cardiac fibroblasts after angiotensin-II stimulation. Mechanistically, ALK4 haploinsufficiency resulted in the suppression of Smad2/3 activity. Conclusion Our results demonstrate that ALK4 haploinsufficiency ameliorates cardiac fibrosis and dysfunction in a mouse pressure-overload model associated with inhibition of cardiac fibroblast activation and cardiomyocyte apoptosis through the suppression of Smad2/3 activity, and suggest that ALK4 is a novel therapeutic target in treating pressure overload-induced cardiac remodeling and heart failure.

CD22 is a surface immunoglobulin implicated in negative regulation of B cell receptor (BCR) signaling; particularly inhibiting intracellular Ca2+ (Ca2+i)signals. Its cytoplasmic tail contains six tyrosine residues (Y773/Y783/Y817/Y828/Y843/Y863, designated Y1~Y6 respectively), including three (Y2/5/6) lying within immunoreceptor tyrosine-based inhibitory motifs (ITIMs) that serve to recruit the protein tyrosine phosphatase SHP-1 after BCR activation-induced phosphorylation. The mechanism of inhibiting Ca2+i by CD22 has been poorly understood. Previous study demonstrated that CD22 associated with plasma membrane calcium-ATPase (PMCA) and enhanced its activity (Chen, J. et al. Nat Immunol 2004;5:651-7). The association is dependent on BCR activation-induced cytoplasmic tyrosine phosphorylation, because CD22 with either all six tyrosines mutated to phenylalanines or cytoplasmic tail truncated loses its ability to associate with PMCA. However, which individual or a group of tyrosine residues determine the association and how CD22 and PMCA interacts, are still unclear. In this study, by using a series of CD22 tyrosine mutants, we found that ITIM Y2/5/6 accounts for 34.3~37.1% Ca2+i inhibition but is irrelevant for CD22/PMCA association. Non-ITIM Y4 and its YEND motif contribute to the remaining 69.4~71.7% Ca2+i inhibition and is the binding site for PMCA-associated Grb2. Grb2, independently of BCR cross-linking, is constitutively associated with and directly binds to PMCA in both chicken and human B cells. Knockout of Grb2 by CRISPR/Cas9 completely disrupted the CD22/PMCA association. Thus, our results demonstrate for the first time that in addition to previously-identified ITIM/SHP-1-dependent pathway, CD22 holds a major pathway of negative regulation of Ca2+i signal, which is ITIM/SHP-1-independent, but Y4/Grb2/PMCA-dependent.

In comparison to various power generating devices/energy conversion systems, solid oxide fuel cell (SOFC) has received much attention due to its remarkable efficiency, reliability and low pollution. Modified pechini method is used to synthesize La0.7Sr0.3Ti0.1Fe0.6Ni0.3O3-δ (LSTFN) perovskite material and is utilized as cathode and anode for intermediate-temperature symmetrical solid oxide fuel cell (IT-SSOFC). The X-ray diffractometer (XRD) patterns reveals perovskite cubic structure with single phase for LSTFN. The maximum conductivity ~318 Scm−1 is obtained for LSTFN in air at a temperature of 700 °C, but is decreased with further increase in temperature due to small polaron mechanism. However, the conductivity is enhanced up to ~1.1 Scm−1 at the same temperature in the presence of dry H2, being capable to be utilized both as anode and cathode. Low polarization resistance (Rp) ~0.047 (with air) and 0.201Ωcm2 (with dry H2) is exhibited by as prepared perovskite material at a temperature of 800 °C. The fabricated symmetrical cell maintains good stability with no apparent degradation during the tested hours ~300 in air and ~100 with dry H2.

La0.6Sr0.4Co0.2Fe0.8O3-δ (LSCF) is a promising cathode material for solid oxide fuel cells (SOFCs). Hence, structured LSCF coatings were deposited in this study by atmospheric laminar plasma spraying (ALPS) at the spraying distances from 150 mm to 250 mm. Particles with high temperature were obtained at low plasma arc power of only 15 kW. The microstructural characterization of the coatings showed cluster-like morphologies on the top surfaces and columnar features with large-scale vertical cracks at the cross section. An evolution mechanism of the coatings was proposed based on particle size. The movement of particles during laminar plasma jet depended on particle size, which was studied experimentally and by simulation to explain the selective deposition. The coatings showed lower polarization resistance when compared to LSCF coatings deposited by atmospheric plasma spraying (APS) due to large vertical crack densities. The high output performance of the SOFCs with the LSCF cathode also proved this phenomenon. The results suggest that ALPS provides extensive options for SOFCs manufacturing.

Background Activin receptor‐like kinase 4 ( ALK 4) is highly expressed in mammal heart. Atrial fibrillation ( AF ) is closely related to ventricular pressure overload. Because pressure overload increases atrial pressure and leads to atrial remodeling, it would be informative to know whether ALK 4 exerts potential effects on atrial remodeling and AF vulnerability in a pressure‐overload model. Methods and Results Wild‐type littermates and ALK 4 +/− mice were subjected to abdominal aortic constriction or a sham operation. After 4 or 8 weeks, echocardiographic and hemodynamic measurements were performed, and inducibility of AF was tested. The hearts were divided into atria and ventricles and then were fixed in formalin for staining, or they were weighted and snap‐frozen for quantitative real‐time polymerase chain reaction and Western blot analysis. Compared with wild‐type littermates, ALK 4 +/− mice demonstrated a similar extent of atrial hypertrophy but significantly suppressed atrial fibrosis at 8 weeks post–abdominal aortic constriction. ALK 4 haplodeficiency partially blocked abdominal aortic constriction–induced upregulation of monocyte chemotactic protein 1 and interleukin‐6, and the increased chemotaxin of macrophages. ALK 4 haplodeficiency also blunted a reduction of connexin 40 and redistribution of connexin 43 from the intercalated disk to the lateral membranes, thereby improving localized conduction abnormalities. Meanwhile, ALK 4 haplodeficiency inhibited abdominal aortic constriction–induced decreased I N a , I C a‐L and I K 1 densities as well as the accompanying action potential duration shortening. Mechanistically, ALK 4 haploinsufficiency resulted in the suppression of Smad2/3 activity in this model. Conclusions Our results demonstrate that ALK 4 haplodeficiency ameliorates atrial remodeling and vulnerability to AF in a pressure‐overload model through inactivation of the Smad2/3 pathway, suggesting that ALK 4 might be a potential therapeutic target in combating pressure overload–induced AF .

Cardiac fibrosis (CF), a process characterized by potentiated proliferation of cardiac fibroblasts and excessive secretion and deposition of extracellular matrix (ECM) from the cells, contributes strongly to the pathogenesis of a series of cardiovascular (CV) diseases, including AMI, heart failure and atrial fibrillation. Endothelial-mesenchymal transition (EndMT), one of the sources of transformed cardiac fibroblasts, has been reported as a key factor involved in CF. However, the molecular basis of EndMT has not been thoroughly elucidated to date. At the posttranscriptional level, of the three epigenetic regulators, writer and eraser are reported to be involved in EndMT, but the role of reader in the process is still unknown. In this study, we aimed to explore the role of Bromodomain-containing protein 4 (BRD4), an acetyl-lysine reader protein, in EndMT-induced CF and related mechanisms. We found that BRD4 was upregulated in endothelial cells (ECs) in the pressure-overload mouse heart and that its functional inhibitor JQ1 potently attenuated the TAC-induced CF and preserved cardiac function. In umbilical vein endothelial cells (HUVECs) and mouse aortic endothelial cells (MAECs), bothJQ1 and shRNA-mediated silencing of BRD4 blocked TGF-β-induced EC migration, EndMT and ECM synthesis and preserved the EC sprouting behavior, possibly through the downregulation of a group of transcription factors specific for EndMT (Snail, Twist and Slug), the Smads pathway and TGF-β receptor I. In the absence of TGF-β stimulation, ectopic expression of BRD4 alone could facilitate EndMT, accelerate migration and increase the synthesis of ECM. In vivo, JQ1 also attenuated TAC-induced EndMT and CF, which was consistent with JQ1's intracellular mechanisms of action. Our results showed that BRD4 plays a critical role in EndMT-induced CF and that targeting BRD4 might be a novel therapeutic option for CF.

High manufacturing costs and long-term degradation are the main problems that have become a “bottleneck” and impeded SOFC’s further development. It is well known that a high operating temperature is the major cause that leads to these problems. As such, reducing the operating temperature becomes a hotspot of research. It has been reported that a uniform and dense coating can be prepared by using very low pressure plasma spraying (VLPPS) technology. The current study focuses on VLPPS for application in large-area (~100 × 100 mm) porous metal supported solid oxide fuel cell (MSSOFC), especially for the preparation of the electrolyte. It was found that the densification of the electrolyte was very good, as confirmed by the open-circuit voltage (OCV) of the cell. In the temperature range of 550~750°C, the OCV of the cell stabilized between 1.05 V and 1.1 V. The power density of the cell was also measured.

BACKGROUND AND PURPOSE: 9‐Phenanthrol, known as a specific inhibitor of the transient receptor potential melastatin 4 (TRMP4) channel, has been shown to modulate cardiac electrical activity and exert antiarrhythmic effects. However, its pharmacological effects remain to be fully explored. Here, we tested the hypothesis that cardiac sodium current inhibition contributes to the cardioprotective effect of 9‐phenanthrol. EXPERIMENTAL APPROACH: Single ventricular myocytes (VMs) and Purkinje cells (PCs) were enzymatically isolated from rabbits. Arterially perfused rabbit wedge preparations were also used, and transmural electrocardiogram and endocardial action potentials (APs) were simultaneously recorded. Wild‐type and mutated human recombinant SCN5A were expressed in HEK293 cells. Anemonia toxin II (ATX‐II) was used to amplify the late sodium current (I(NaL)) and induce arrhythmias. Whole‐cell patch clamp technique was used to record APs and ionic currents. KEY RESULTS: 9‐Phenanthrol (10–50 μM) stabilized ventricular repolarization and abolished arrhythmias induced by ATX‐II in both isolated VMs, PCs and wedge preparations. Further study revealed that 9‐phenanthrol modulated the gating properties of cardiac sodium channels and dose‐dependently inhibited I(NaL) and peak sodium current (I(NaP)) in VMs with an IC(50) of 18 and 71.5 μM respectively. Its ability to inhibit I(NaL) was further confirmed in PCs and HEK293 cells expressing SCN5A mutations. CONCLUSIONS AND IMPLICATIONS: Our results indicate that 9‐phenanthrol modulates the gating properties of cardiac sodium channels and inhibits I(NaL) and I(NaP), which may contribute to its antiarrhythmic and cardioprotective effects.

Background: Phospholemman (PLM) is an important phosphorylation substrate for protein kinases A and C in the heart. Until now, the association between PLM phosphorylation status and L-type calcium channels (LTCCs) gating has not been fully understood. We investigated the kinetics of LTCCs in HEK 293T cells expressing phosphomimetic or nonphosphorylatable PLM mutants. Methods: The LTCCs gating was measured in HEK 293T cells transfected with LTCC and wild-type (WT) PLM, phosphomimetic or nonphosphorylatable PLM mutants: 6263AA, 6869AA, AAAA, 6263DD, 6869DD or DDDD. Results: WT PLM significantly slowed LTCCs activation and deactivation while enhanced voltage-dependent inactivation (VDI). PLM mutants 6869DD and DDDD significantly increased the peak of the currents. 6263DD accelerated channel activation, while 6263AA slowed it more than WT PLM. 6869DD significantly enhanced PLM-induced increase of VDI. AAAA slowed the channel activation more than 6263AA, and DDDD accelerated the channel VDI more than 6869DD. Conclusions: Our results demonstrate that phosphomimetic PLM could stimulate LTCCs and alter their dynamics, while PLM nonphosphorylatable mutant produced the opposite effects.

Small GTPase Rac1 is a member of the Ras superfamily, which plays important roles in regulation of cytoskeleton reorganization, cell growth, proliferation, migration, etc. The aim of this study was to determine how a constitutively active Rac1b regulated cell proliferation and to investigate the effects of the Rac1b inhibitor sanguinarine.Three HEK293T cell lines stably overexpressing GFP, Rac1-GFP or Rac1b-GFP were constructed by lentiviral infection. The cells were treated with sanguinarine (1 μmol/L) or its analogue berberine (1 μmol/L) for 4 d. Cell proliferation was evaluated by counting cell numbers and with a BrdU incorporation assay. The levels of cleaved PARP-89 (an apoptosis marker) and cyclin-D1 (a proliferative index) were measured using Western blotting.In 10% serum-containing media, overexpressing either Rac1 or Rac1b did not significantly change the cell proliferation. In the serum-starved media, however, the survival rate of Rac1b cells was significantly increased, whereas that of Rac1 cells was moderately increased. The level of cleaved PARP-89 was significantly increased in serum-starved Rac1 cells, but markedly reduced in serum-starved Rac1b cells. The level of cyclin-D1 was significantly increased in both serum-starved Rac1 and Rac1b cells. Treatment with sanguinarine, but not berberine, inhibited the proliferation of Rac1b cells, which was accompanied by significantly increased the level of PARP-89, and decreased both the level of cyclin-D1 and the percentage of BrdU positive cells.Rac1b enhances the cell proliferation under a growth-limiting condition via both anti-apoptotic and pro-proliferative mechanisms. Sanguinarine, as the specific inhibitor of Rac1b, is a potential therapeutic agent for malignant tumors with up-regulated Rac1b.

Rate-dependent repolarization (RDR) of action potential (AP) in cardiomyocyte plays a critical role in the genesis of arrhythmias and RDR in atrium has been linked with atrial fibrillation. However, detailed studies focusing on the role of RDR in rabbit atrium are scant. In this study, atrial cells were isolated from rabbit heart and rate-dependent property was explored in single atrial cell to elucidate the underlying mechanism. Our results indicated that rate-dependent prolongation was evident at the action potential duration at 20% (APD20) and 50% (APD50) repolarization but not at 90% repolarization (APD90) under control condition. Using transient outward potassium current (I

Atrial fibrosis, the hallmark of structural remodeling associated with atrial fibrillation (AF), is characterized by abnormal proliferation of atrial fibroblasts and excessive deposition of extracellular matrix. Transforming growth factor-β1 (TGF-β1)/activin receptor-like kinase 5 (ALK5)/Smad2/3/4 pathway has been reported to be involved in the process. Recent studies have implicated both activin A and its specific downstream component activin receptor-like kinase 4 (ALK4) in stimulating fibrosis in non-cardiac organs. We recently reported that ALK4 haplodeficiency attenuated the pressure overload- and myocardial infarction-induced ventricular fibrosis. However, the role of activin A/ALK4 in the pathogenesis of atrial fibrosis and vulnerability to AF remains unknown. Our study provided experimental and clinical evidence for the involvement of activin A and ALK4 in the pathophysiology of atrial fibrosis and AF. Patients with AF had higher activin A and ALK4 expression in atriums as compared to individuals devoid of AF. After angiotensin-II (Ang-II) stimulation which mimicked atrial fibrosis progression, ALK4-deficient mice showed lower expression of ALK4 in atriums, reduced activation of atrial fibroblasts, blunted atrial enlargement and atrial fibrosis, and further reduced AF vulnerability upon right atrial electrophysiological studies as compared to wild-type littermates. Moreover, we found that apart from the well-known TGF-β1/ALK5 pathway, the activation of activin A/ALK4/smad2/3 pathway played an important role in the pathogenesis of Ang-II-mediated atrial fibrosis and inducibility of AF, suggesting that targeting ALK4 might be a potential therapy for atrial fibrosis and AF.

Background Purkinje cells (PCs) have a steeper rate dependence of repolarization and are more susceptible to arrhythmic activity than do ventricular myocytes (VMs). Late sodium current (I NaL ) is rate dependent and contributes to rate dependence of repolarization. Objective This study sought to test our hypothesis that PCs have a larger rate dependence of I NaL , contributing to their steeper rate dependence of repolarization and higher susceptibility to arrhythmic activity, than do VMs. Methods I NaL was recorded in isolated rabbit PCs and VMs with the whole-cell patch clamp technique. Action potential was examined using the microelectrode technique. Results Compared with VMs, PCs exhibited a significantly larger rate dependence of I NaL with a larger I NaL to basic cycle length (BCL) slope. Moreover, PCs had a larger rate dependence of I NaL decay and slower recovery kinetics. Interestingly, the larger rate dependence of I NaL matched to a steeper rate dependence of action potential duration (APD) in PCs. The I NaL blocker tetrodotoxin significantly blunted, while the I NaL enhancer anemone toxin (ATX-II) significantly increased, the rate dependence of I NaL and APD in PCs and VMs. In the presence of ATX-II, the rate dependence of I NaL in PCs was markedly larger than that in VMs, causing a much steeper rate dependence of APD in PCs. Accordingly, PCs exhibited greater rate-dependent electrical instability and were more prone to ATX-II–induced early afterdepolarizations, which were completely inhibited by the I NaL inhibitor ranolazine. Conclusion PCs have a significantly larger rate dependence of I NaL than do VMs because of distinctive I NaL decay and recovery kinetics, which contributes to their larger rate adaptation, and simultaneously predisposes them to a higher risk of arrhythmogenesis.

A discrete assimilation system for a one-dimensional variable coefficient convection–diffusion equation is constructed. The variational adjoint method combined with the regularization technique is employed to retrieve the initial condition and diffusion coefficient with the aid of a set of simulated observations. Several numerical experiments are performed: (a) retrieving both the initial condition and diffusion coefficient jointly (Experiment JR), (b) retrieving either of them separately (Experiment SR), (c) retrieving only the diffusion coefficient with the iteration count increased to 800 (Experiment NoR-SR), and (d) retrieving only the diffusion coefficient with the consideration of a regularization term based on the Experiment NoR-SR (Experiment AdR-SR). The results indicate that within the limit of 100 iterations, the retrieval quality of the Experiment SR is better than those from the Experiment JR. Compared with the initial condition, the diffusion coefficient is a little difficult to retrieve, whereas we still achieve the desired result by increasing the iterations or integrating the regularization term into the cost functional for the improvement with respect to the diffusion coefficient. Further comparisons between the Experiment NoR-SR and AdR-SR show that the regularization term can really help not only improve the precision of retrieval to a large extent, but also speed up the convergence of solution, even if some perturbations are imposed on those observations.

Some new generalized solitary solutions of the Klein–Gordon–Schrödinger equations are obtained using the Exp-function method, which include some known solutions obtained by the F-expansion method and the homogeneous balance method in the open literature as special cases. It is shown that the Exp-function method is a straight, concise, reliable and promising mathematical tool for solving nonlinear evolution equations arising in mathematical physics.

The C k (k ≥ 3) stability of nonlinear barotropic vorticity equation was proved. The necessary and sufficient conditions for the initial value problem to be well-posed were presented. Under the conditions of well-posedness, the corresponding analytical solution was also gained.

On condition that the basic equations set of atmospheric motion possesses the best stability in the smooth function classes, the structure of solution space for local analytical solution is discussed, by which the third-class initial value problem with typicality and application is analyzed. The calculational method and concrete expressions of analytical solution about the well-posed initial value problem of the third kind are given in the analytic function classes. Near an appointed point, the relevant theoretical and computational problems about analytical solution of initial value problem are solved completely in the meaning of local solution. Moreover, for other type of problems for determining solution, the computational method and process of their stable analytical solution can be obtained in a similar way given in this paper.

Rac1b is a constitutively activated, alternatively spliced form of the small GTPase Rac1. Previous studies showed that Rac1b promotes cell proliferation and inhibits apoptosis. In the present study, we used microarray analysis to detect genes differentially expressed in HEK293T cells and SW480 human colon cancer cells stably overexpressing Rac1b. We found that the pro-proliferation genes JNK2, c-JUN and cyclin-D1 as well as anti-apoptotic AKT2 and MCL1 were all upregulated in both lines. Rac1b promoted cell proliferation and inhibited apoptosis by activating the JNK2/c-JUN/cyclin-D1 and AKT2/MCL1 pathways, respectively. Very low Rac1b levels were detected in the colonic epithelium of wild-type Sprague-Dawley rats. Knockout of the rat Rac1 gene exon-3b or knockdown of endogenous Rac1b in HT29 human colon cancer cells downregulated only the AKT2/MCL1 pathway. Our study revealed that very low levels of endogenous Rac1b inhibit apoptosis, while Rac1b upregulation both promotes cell proliferation and inhibits apoptosis. It is likely the AKT2/MCL1 pathway is more sensitive to Rac1b regulation.

Dual-specificity phosphatase 14 (Dusp14), an important negative modulator of mitogen-activated protein kinase (MAPK) signaling pathways, has been implicated in inflammatory immune response, cancers, cell differentiation and proliferation. The role of Dusp14 in chronic pressure overload-induced cardiac hypertrophy has not been explored. Here we have shown that Dusp14-/- knockout mice and cardiac-specific Dusp14 transgenic mice were generated and subjected to aortic banding (AB) for 4 weeks. Our results demonstrated that genetic loss of Dusp14 significantly aggravated cardiac hypertrophy, fibrosis, ventricular dilation and dysfunction, whereas transgenic cardiac-specific Dusp14 overexpression significantly attenuated AB-induced cardiac dysfunction and remodeling. In vitro, adenoviral overexpression of constitutive Dusp14 blocked angiotensin II-induced hypertrophic growth of cardiomyocytes, while Dusp14 knockdown led to opposite effects. Mechanistically, excessive phosphorylation of TAK1, P38MAPK and JNK1/2 was evidenced in Dusp14-/- knockout mice post-AB and inactivation of TAK1-P38MAPK and -JNK1/2 signaling using TAK1 inhibitor 5Z-7-ox shares similar antihypertrophic effect as Dusp14 overexpression. Moreover, we show that Dusp14 directly interacted with TAK1. Results from present experiments indicate that Dusp14 protects the heart from AB-induced cardiac hypertrophy and dysfunction possibly through inactivation of TAK1-P38MAPK/-JNK1/2 signaling pathway. Future studies are warranted to test the feasibility of overexpressing Dusp14 as a therapeutic strategy to attenuate cardiac hypertrophy and failure.