The extracellular cyclophilin A-integrin ß2 complex as a therapeutic target of viral pneumonia.

The acute respiratory virus infection can induce uncontrolled inflammatory responses, such as cytokine storm and viral pneumonia, which are the major causes of death in clinical cases. Cyclophilin A (CypA) is mainly distributed in the cytoplasm of resting cells and released into the extracellular space in response to inflammatory stimuli. Extracellular CypA (eCypA) is upregulated and promotes inflammatory response in severe COVID-19 patients. However, how eCypA promotes virus-induced inflammatory response remains elusive. Here, we observe that eCypA is induced by influenza A and B viruses and SARS-CoV-2 in cells, mice, or patients. Anti-CypA mAb reduces pro-inflammatory cytokines production, leukocytes infiltration, and lung injury in virus-infected mice. Mechanistically, eCypA binding to integrin ß2 triggers integrin activation, thereby facilitating leukocyte trafficking and cytokines production via the focal adhesion kinase (FAK)/GTPase and FAK/ERK/P65 pathways, respectively. These functions are suppressed by the anti-CypA mAb that specifically blocks eCypA-integrin ß2 interaction. Overall, our findings reveal that eCypA-integrin ß2 signaling mediates virus-induced inflammatory response, indicating that eCypA is a potential target for antibody therapy against viral pneumonia.

Risk factors associated with recurrence of Henoch-Schonlein purpura: a retrospective study.

Background: Recurrence is considered a vital problem for assessing the prognosis of Henoch-Schonlein purpura (HSP). The objective of this study was to evaluate factors affecting the recurrence in children with HSP. Methods: We retrospectively reviewed records of 368 patients under the age of 16 years diagnosed with HSP from October 2019 to December 2020 in Beijing Children's Hospital. Patients were divided into a non-recurrence group and a recurrence group according to whether there was a recurrence. Incidence of manifestation, possible cause, age, and treatment were retrospectively analyzed. Univariate and multivariate logistic regression analyses were used to determine the risk factors of recurrence in HSP. Results: Percentages of patients were 65.2% for the non-recurrence group and 34.8% for the recurrence group. The percentage of patients with renal involvement was significantly higher in the recurrence group (40.6%) than in the non-recurrence group (26.3%). Respiratory tract infection was the most frequent trigger: 67.5% in the non-recurrence group and 66.4% in the recurrence group. Recurrence was more likely to occur in patients aged >6 years (53.3% vs. 71.9%). Logistic regression analysis revealed that hematuria plus proteinuria was an independent risk factor for the recurrence of HSP. Conversely, animal protein, exercise restriction, and age ≤6 years were independent favorable factors for the non-recurrence of HSP. Conclusion: These results suggest that organ involvement, exercise, and diet management during the initial episode of HSP should be strictly monitored for children with HSP. Adequate clinical intervention for these risk factors may limit or prevent HSP recurrence. Moreover, renal involvement is associated with the long-term prognosis of HSP.

Dan Bai Xiao Formula combined with glucocorticoids and cyclophosphamide for pediatric lupus nephritis: A pilot prospective study.

BACKGROUND: Patients with lupus nephritis (LN) typically undergo long-term treatment with glucocorticoids (GCs) and immunosuppressants. There is a growing demand for optimal therapy with better remission results and fewer side effects. Sustained traditional Chinese medicine (TCM) might be quite valuable for multitarget therapy, reducing the total dosage of GCs and minimizing the side effects of immunosuppressants. AIM: To evaluate whether Dan Bai Xiao Formula (DBXF) can reduce the exposure to GCs and cyclophosphamide (CYC) and to assess the efficacy and safety of DBXF for the resolution of proteinuria and hematuria in children with LN. METHODS: A 24-wk pilot study was conducted at Beijing Children's Hospital. Children with active LN were divided into either a TCM group or a control group. Children in the TCM group received DBXF combined with GCs and CYC, and the ones in the control group received GCs and CYC every 4 wk for 24 wk. The primary endpoints of this trial were urinary protein excretion of < 150 mg/d and normal serum albumin concentration and renal function. RESULTS: The trial included 78 children, of whom 38 received GCs and CYC treatment (control group) and the remaining 40 received DBXF combined with GCs and CYC treatment (TCM group). At week 24, the TCM group showed a better rate of complete remission (42.5%); however, there was no significant difference compared with the control group (31.5%, P > 0.05). The urine red blood cell count and urine protein level were significantly lower in the TCM group than in the control group at weeks 4, 12, and 24 (P < 0.05). Furthermore, patients in the TCM group had a lower proportion of methylprednisolone pulses than those in the control group (1.30 ± 1.41 vs 3.05 ± 2.02, P < 0.0001). The ending GC dose was significantly lower in the TCM group than in the control group (P < 0.001). Moreover, more hepatic function damage, gastrointestinal adverse effects, and hypertension were observed in the control group than in the TCM group (P < 0.05). CONCLUSION: The findings suggest that DBXF treatment is effective and safe as a supplementary therapy for LN and is superior to routine GC and CYC therapy. DBXF containing combination treatment possibly results in a faster resolution of proteinuria and hematuria, smoother GC reduction, fewer methylprednisolone pulses, and fewer adverse events.

Construction of cationic polyfluorinated azobenzene/reduced graphene oxide for simultaneous determination of dopamine, uric acid and ascorbic acid.

A highly sensitive cationic polyfluorinated azobenzene/reduced graphene oxide (C3F7-azo+/RGO) nanocomposite electrochemical sensor for simultaneous detection of dopamine (DA), ascorbic acid (AA) and uric acid (UA) was successfully synthesized using a facile exfoliation/restacking method. The nanocomposite is self-assembled from oppositely charged graphene oxide nanosheets (GO) and polyfluorinated azobenzene cations (C3F7-azo+), and then obtained by electrochemical reduction. The structure and electrochemical properties were characterized by X-ray diffraction (XRD), energy dispersive spectrometer analysis (EDS), transmission electron microscope (TEM) and scanning electron microscope (SEM). The electrochemical property of C3F7-azo+/RGO was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). It can be clearly seen from experimental results that C3F7-azo+/RGO-modified electrode (C3F7-azo+/RGO/GCE) can detect DA, AA and UA simultaneously, and has good stability and anti-interference performance. The detection limits are 65 nM, 8 nM and 11 nM for DA, AA and UA in the ranges 57.28-134.28 µM, 0.04-6.01 µM, 9.23-23.45 µM, respectively.

Effect of animal protein diet on the prognosis of children with Henoch-Schönlein purpura. / 动物蛋白饮食对过敏性紫癜患儿预后的影响分析.

OBJECTIVES: To study the association of animal protein diet with the recurrence of Henoch-Schönlein purpura (HSP)/skin rash and the risk factors for recurrence of HSP. METHODS: A prospective analysis was performed for 121 children with HSP who were admitted to the Beijing Children's Hospital from October to December 2020. The children were given the doctor's advice of the same diet (animal protein diet could be added after 1 week without new-onset skin rash). Follow-up was performed at the outpatient service for half a year. According to the presence or absence of animal protein intake, the children were divided into an observation group with 65 children and a control group with 56 children. The times of skin rash recurrence, the incidence of HSP recurrence, and the incidence of kidney injury were compared between the two groups. According to the presence or absence of recurrence, the children were divided into a recurrence group with 32 children and a non-recurrence group with 89 children. A questionnaire on food frequency was used to record the daily intake of animal protein in the two groups. A multivariate logistic regression analysis was used to identify the risk factors for recurrence of HSP in children. RESULTS: There was no significant difference between the observation and control groups in the times of skin rash recurrence, the incidence rate of HSP recurrence, and the incidence rate of kidney injury (P>0.05). There was no significant difference in the daily intake of animal protein between the recurrence and non-recurrence groups (P>0.05). The multivariate logistic regression analysis showed that presence of kidney injury at initial onset, respiratory infection after cure for the first time, and lack of exercise control after cure for the first time were independent risk factors for the recurrence of HSP in children (P<0.05). CONCLUSIONS: There is no significant association between animal protein diet and the recurrence of HSP or skin rash. Timely treatment of kidney injury, avoidance of infection after cure, and limitation of strenuous exercise may help to reduce the recurrence rate of HSP in children. Citation.

Automatic 3D adaptive vessel segmentation based on linear relationship between intensity and complex-decorrelation in optical coherence tomography angiography.

BACKGROUND: Vascular quantitative metrics have been widely used in the preclinical studies and clinical applications (e.g., the diagnosis and treatment of port wine stain, PWS), which require accurate vessel segmentation. An automatic 3D adaptive vessel segmentation is in need for a reproducible and objective quantification of the optical coherence tomography angiography (OCTA) image. METHODS: Human skin imaging was performed with a lab-built optical coherence tomography (OCT) system. Rather than separately applying the conventional 2-step (intensity and binarization) thresholding in the decorrelation-contrast OCTA, we proposed a 3D adaptive threshold using the linear relationship between the local intensity and complex-decorrelation which was termed as inverse SNR-decorrelation (ID) threshold. Furthermore, the ID threshold was automatically determined by defining a binary image similarity (BISIM) index as the feedback and searching the ID threshold with the minimal BISIM value. The proposed ID-BISIM threshold was applied to the acquired OCTA skin images for further vessel quantification. RESULTS: The proposed ID-BISIM threshold enabled a 3D adaptive binarization and presented superior sensitivity and specificity in vessel segmentation over conventional 2-step thresholding method in the decorrelation-contrast OCTA and a 37-65% improvement of the Youden's index in human skin experiments. The 3D binarization enabled a depth-resolved vessel skeleton and enhanced the differentiation of the overlapping vessels in the depth direction. Using ID-BISIM, the quantitative OCTA image presented a significant increase of vessel diameter index (P=0.0015) and vessel area density (VAD) (P=0.0485) as well as a significant decrease of vessel complexity index (VCI) (P=0.0094) in PWS lesion skin compared with normal skin. CONCLUSIONS: The proposed ID-BISIM method enables an automatic 3D adaptive vessel segmentation with enhanced performance in quantitative OCTA. The vascular quantitative metrics would be a useful tool for improving the diagnosis and the treatment of PWS.

Natriuretic peptides and Forkhead O transcription factors act in a cooperative manner to promote cardiomyocyte cell cycle re-entry in the postnatal mouse heart.

BACKGROUND: Cardiomyocytes proliferate rapidly during fetal life but lose their ability of proliferation soon after birth. However, before terminal withdrawal from the cell cycle, cardiomyocytes undergo another round of cell cycle during early postnatal life in mice. While a transient wave of increased DNA synthesis in cardiomyocyte has been observed in postnatal mouse hearts, the molecular mechanisms describing cardiomyocyte cell cycle re-entry remain poorly understood. Atrial and B-type natriuretic peptides (ANP and BNP) are abundantly expressed in embryonic heart ventricles. After birth, the expression of both genes is strongly reduced in the ventricular myocardium. Forkhead O (FOXO) transcription factors are expressed in both embryonic and postnatal heart ventricles. Their transcriptional activity negatively affects cardiomyocyte proliferation. Upon phosphorylation, FOXO is translocated to the cytoplasm and is transcriptionally inactive. Despite these important findings, it remains largely unknown whether natriuretic peptides and FOXO cooperatively play a role in regulating cardiomyocyte cell cycle activity during early postnatal life. RESULTS: We observed that the expression of ANP and BNP and the level of phosphorylated FOXO were transiently increased in the postnatal mouse heart ventricles, which coincided with the burst of cardiomyocyte cell cycle re-entry during early postnatal life in mice. Cell culture studies showed that ANP/BNP signaling and FOXO cooperatively promoted cell cycle activity in neonatal mouse cardiomyocytes. The enhanced cell cycle activity observed in combined treatment of ANP/BNP and dominant-negative FOXO (DN-FOXO), which can bind FOXO recognition sites on DNA but cannot activate transcription, was primarily mediated through natriuretic peptide receptor 3 (Npr3). In mice, simultaneous application of ANP and DN-FOXO in postnatal hearts reactivated cell cycle in cardiomyocytes, resulting in reduced scar formation after experimental myocardial infarction. CONCLUSIONS: Our data demonstrate the cooperative effects of natriuretic peptide and DN-FOXO on promoting cardiomyocyte cell cycle activity and mouse cardiac repair and regeneration after injury.

High performance OCTA enabled by combining features of shape, intensity, and complex decorrelation.

Motion contrast optical coherence tomography angiography (OCTA) entails a precise identification of dynamic flow signals from the static background, but an intermediate region with voxels exhibiting a mixed distribution of dynamic and static scatterers is almost inevitable in practice, which degrades the vascular contrast and connectivity. In this work, the static-dynamic intermediate region was pre-defined according to the asymptotic relation between inverse signal-to-noise ratio (iSNR) and decorrelation, which was theoretically derived for signals with different flow rates based on a multi-variate time series (MVTS) model. Then the ambiguous voxels in the intermediate region were further differentiated using a shape mask with adaptive threshold. Finally, an improved OCTA classifier was built by combining shape, iSNR, and decorrelation features, termed as SID-OCTA, and the performance of the proposed SID-OCTA was validated experimentally through mouse retinal imaging.

Improvement of Decorrelation-Based OCT Angiography by an Adaptive Spatial-Temporal Kernel in Monitoring Stimulus-Evoked Hemodynamic Responses.

Complex decorrelation-based OCT angiography (OCTA) has the potential for monitoring hemodynamic activities in a label-free, high-resolution, and quantitative manner. To improve the measurement dynamic range and uncertainty of blood flow, an adaptive spatial-temporal (ST) kernel was proposed for decorrelation estimation and it was validated through a theoretical simulation and experimental measurements. The ensemble size in the decorrelation computation was effectively enlarged by collecting samples of the phasor pair in both the spatial and temporal dimensions. The spatial sub-kernel size was adaptively changed to suppress the influence of bulk motion in the temporal dimension by solving a maximum entropy model. Using the flow phantom, it was observed that the decorrelation dynamic range presented an increase of ~49% and the uncertainty exhibited a decrease of ~40% and ~38% in the saturation and background limits, respectively. In monitoring the stimulus-evoked hemodynamic response, the extended dynamic range enabled an improvement of ~180% in the separability between different stimulation modes. Furthermore, the suppressed uncertainty and motion artifacts allowed a reliable temporal analysis of the hemodynamic response. The proposed adaptive ST-kernel will greatly promote the development of decorrelation-based quantitative OCTA in hemodynamic studies.

Research Status and Development Trend of MEMS Switches: A Review.

MEMS switch is a movable device manufactured by means of semiconductor technology, possessing many incomparable advantages such as a small volume, low power consumption, high integration, etc. This paper reviews recent research of MEMS switches, pointing out the important performance indexes and systematically summarizing the classification according to driving principles. Then, a comparative study of current MEMS switches stressing their strengths and drawbacks is presented, based on performance requirements such as driven voltage, power consumption, and reliability. The efforts of teams to optimize MEMS switches are introduced and the applications of switches with different driving principles are also briefly reviewed. Furthermore, the development trend of MEMS switch and the research gaps are discussed. Finally, a summary and forecast about MEMS switches is given with the aim of providing a reference for future research in this domain.

Stachydrine hydrochloride alleviates pressure overload-induced heart failure and calcium mishandling on mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicine Leonurus japonicus Houtt. has a long history in the treatment of cardiovascular diseases. Stachydrine hydrochloride, the main bioactive ingredient extracted from Leonurus japonicus Houtt., has been shown to have cardioprotective effects. However, the underlying mechanisms of stachydrine hydrochloride haven't been comprehensively studied so far. AIM OF THE STUDY: The aim of this study was to investigate the protective role of stachydrine hydrochloride in heart failure and elucidate its possible mechanisms of action. MATERIALS AND METHODS: In vivo, transverse aorta constriction was carried out in C57BL/6J mice, and thereafter, 7.2 mg/kg telmisartan (a selective AT1R antagonist as positive control) and 12 mg/kg stachydrine hydrochloride was administered daily intragastrically for 4 weeks. Cardiac function was evaluated by assessing morphological changes as well as echocardiographic and haemodynamic parameters. In vitro, neonatal rat cardiomyocytes or adult mice cardiomyocytes were treated with stachydrine hydrochloride and challenged with phenylephrine (α-AR agonist). Ventricular myocytes were isolated from the hearts of C57BL/6J mice by Langendorff crossflow perfusion system. Intracellular calcium was measured by an ion imaging system. The length and movement of sarcomere were traced to evaluate the systolic and diastolic function of single myocardial cells. RESULTS: Stachydrine hydrochloride improved the cardiac function and calcium transient amplitudes, and inhibited the SR leakage and the amount of sparks in cardiac myocytes isolated from TAC mice. We also demonstrated that stachydrine hydrochloride could ameliorated phenylephrine-induced enhance in sarcomere contraction, calcium transients and calcium sparks. Moreover, our data shown that stachydrine hydrochloride blocked the hyper-phosphorylation of CaMKII, RyR2, PLN, and prevented the disassociation of FKBP12.6 from RyR2. CONCLUSION: Our results suggest that stachydrine hydrochloride exerts beneficial therapeutic effects against heart failure. These cardioprotective effects may be associated with the regulation of calcium handling by stachydrine hydrochloride through inhibiting the hyper-phosphorylation of CaMKII.

Fatty Acid Oxidation Promotes Cardiomyocyte Proliferation Rate but Does Not Change Cardiomyocyte Number in Infant Mice.

Cardiomyocyte proliferation accounts for the increase of cardiac muscle during fetal mammalian heart development. Shortly after birth, cardiomyocyte transits from hyperplasia to hypertrophic growth. Here, we have investigated the role of fatty acid ß-oxidation in cardiomyocyte proliferation and hypertrophic growth during early postnatal life in mice. A transient wave of increased cell cycle activity of cardiomyocyte was observed between postnatal day 3 and 5, that proceeded as cardiomyocyte hypertrophic growth and maturation. Assessment of cardiomyocyte metabolism in neonatal mouse heart revealed a myocardial metabolic shift from glycolysis to fatty acid ß-oxidation that coincided with the burst of cardiomyocyte cell cycle reactivation and hypertrophic growth. Inhibition of fatty acid ß-oxidation metabolism in infant mouse heart delayed cardiomyocyte cell cycle exit, hypertrophic growth and maturation. By contrast, pharmacologic and genetic activation of PPARα, a major regulator of cardiac fatty acid metabolism, induced fatty acid ß-oxidation and initially promoted cardiomyocyte proliferation rate in infant mice. As the cell cycle proceeded, activation of PPARα-mediated fatty acid ß-oxidation promoted cardiomyocytes hypertrophic growth and maturation, which led to cell cycle exit. As a consequence, activation of PPARα-mediated fatty acid ß-oxidation did not alter the total number of cardiomyocytes in infant mice. These findings indicate a unique role of fatty acid ß-oxidation in regulating cardiomyocyte proliferation and hypertrophic growth in infant mice.

Yap/Taz regulate alveolar regeneration and resolution of lung inflammation.

Alveolar epithelium plays a pivotal role in protecting the lungs from inhaled infectious agents. Therefore, the regenerative capacity of the alveolar epithelium is critical for recovery from these insults in order to rebuild the epithelial barrier and restore pulmonary functions. Here, we show that sublethal infection of mice with Streptococcus pneumoniae, the most common pathogen of community-acquired pneumonia, led to exclusive damage in lung alveoli, followed by alveolar epithelial regeneration and resolution of lung inflammation. We show that surfactant protein C-expressing (SPC-expressing) alveolar epithelial type II cells (AECIIs) underwent proliferation and differentiation after infection, which contributed to the newly formed alveolar epithelium. This increase in AECII activities was correlated with increased nuclear expression of Yap and Taz, the mediators of the Hippo pathway. Mice that lacked Yap/Taz in AECIIs exhibited prolonged inflammatory responses in the lung and were delayed in alveolar epithelial regeneration during bacterial pneumonia. This impaired alveolar epithelial regeneration was paralleled by a failure to upregulate IκBa, the molecule that terminates NF-κB-mediated inflammatory responses. These results demonstrate that signals governing resolution of lung inflammation were altered in Yap/Taz mutant mice, which prevented the development of a proper regenerative niche, delaying repair and regeneration of alveolar epithelium during bacterial pneumonia.

miR-506 contributes to malignancy of cutaneous squamous cell carcinoma via targeting of P65 and LAMC1.

Previous research has shown that microRNA 506 (miR-506) functions as an essential modulator in the development of many biological reactions, including multiple cancers. However, its involvement in cutaneous squamous cell carcinoma (CSCC) has been rarely reported. In the present work, we investigated the molecular mechanism and function of miR-506 in the regulation of CSCC cell viability and metastasis (migration and invasion). We observed that miR-506 expression was upregulated in both CSCC tissues and cell lines, and that decreased miR-506 expression led to repressed tumorigenesis in CSCC cells. Furthermore, flow cytometry revealed that the depletion of miR-506 resulted in decreased proliferation and increased apoptotic levels in CSCC cells. Meanwhile, it was found that miR-506 decreased CSCC cell migration and invasion in vitro. The dual-luciferase reporter assay also revealed that miR-506 targets the 3'-UTRs of p65 and Laminin C1 (LAMC1) for silencing. Silencing of p65 expression counteracted the pro-apoptotic influence of miR-506 depletion in CSCC cells, while inhibition of LAMC1 expression restored the migration and invasion properties of the CSCC cells. Therefore, the results provide evidence for the need to probe the biological and molecular mechanisms behind the development and progression of CSCC and may lead to novel treatment CSCC strategies.

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

BACKGROUND/AIMS: Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism. METHODS: A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in palmitic acid-stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR. RESULTS: Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes. CONCLUSION: The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.

Buyanghuanwu Decoction alleviated pressure overload induced cardiac remodeling by suppressing Tgf-ß/Smads and MAPKs signaling activated fibrosis.

Buyanghuanwu Decoction (BHD), a traditional Chinese medicine recipe, is a representative prescription for the treatment of qi-deficiency and blood-stasis syndrome. In this study, the effect of BHD on pressure overload induced cardiac remodeling was investigated and possible mechanism underlying was explored. Rats were randomly divided into four groups: sham, transverse aorta constriction (TAC) with saline, TAC with telmisartan (TAC+Tel), and TAC with BHD (TAC+BHD) for 16 weeks (n=6∼8 in each group). Cardiac morphological and functional changes were evaluated by echocardiography and histological methods, the molecular alterations were detected by western blotting. Our results revealed that pressure overload prominently induced cardiac dysfunction, dilated and atrophied left ventricle, decreased cardiomyocyte cross sectional area, and fibrosis. However, BHD, similar to Tel, greatly reversed cardiac dysfunction, left ventricular dilation, and fibrosis, together with increased left ventricular wall thickness and size of cardiomyocyte. Furthermore, activated classical pro-fibrotic signaling of Tgf-ß/Smads and MAPKs after TAC was dramatically suppressed by BHD or Tel treatment. Taken together, it was demonstrated in this study that BHD exerted a cardioprotective effect against pressure overload induced cardiac remodeling via inactivation of Tgf-ß/Smads and MAPKs signaling triggered fibrosis.

Astragaloside-IV Protects Against Heat-induced Apoptosis by Inhibiting Excessive Activation of Mitochondrial Ca2+ Uniporter.

BACKGROUND: Heat causes airway damage during inhalation injury because of bronchial epithelial cell damage. Accumulating evidence shows that mitochondrial uniporter (MCU) is involved in cell damage. We investigated the MCU activity after heat treatment and assessed whether Astragaloside-IV (AS-IV) suppresses heat-induced apoptosis in bronchial epithelial cells by inhibiting the activation of the mitochondrial Ca2+ uniporter (MCU), mitochondrial depolarisation and reactive oxygen species (ROS) production. METHODS: The bronchial epithelial cell line 16HBE14o- was heat treated, and cell apoptosis was induced in vitro and in vivo. AS-IV was inorganically administered to Wistar rats twice a day after thermal inhalation injury, and 16HBE140- cells were treated with AS-IV after incubation at 47°C for 5 min. Protein expression was determined using Western blotting and commercial kits, apoptosis with TUNEL staining, mitochondrial channel activity by patch clamp, reactive oxygen species by MitoSOXTM fluorescence, ATP levels and enzyme activities by commercial kits as well as mitochondrial respiration and calcium by fluorescence. RESULTS: AS-IV markedly inhibited heat-induced apoptosis, as indicated by the increased expression of the pro-apoptotic genes Bak, Bik and Bmf and increased expression of the apoptosis markers Bax, cleaved parp, cleaved caspase3 and cytochrome C. We found that MCU activation promoted mitochondrial Ca2+ overload, ATP depletion, mitochondrial ROS production and cytochrome c release and rapidly induced apoptosis. However, AS-IV treatment reduced excessive MCU activation and led to resistance against mitochondrial Ca2+ overload and excessive cytochrome C release; these effects were blocked by the MCU activator spermine. AS-IV treatment elevated ATP production and decreased ROS activity. CONCLUSIONS: MCU plays crucial roles in heat-induced mitochondrial apoptosis in 16HBE140- cells, suggesting a potential target for AS-IV treatment.

Concurrent CCR7 Overexpression and RelB Knockdown in Immature Dendritic Cells Induces Immune Tolerance and Improves Skin-Graft Survival in a Murine Model.

BACKGROUND/AIMS: Skin transplantation aims to cover skin defects but often fails due to immune rejection of the transplantated tissue. Immature dendritic cells (imDCs) induce immune tolerance but have a low migration rate. After stimulation, imDCs transform into mature DCs, which activate immune rejection. Thus, inducing imDC to obtain a high migration counteracts development of immune tolerance. METHODS & RESULTS: We transfected imDCs with a recombinant adenovirus carrying the CCR7 gene (Ad-CCR7) and a small interfering RNA targeting RelB (RelB-siRNA) to concurrently overexpress CCR7 and downregulate RelB expression. Functionally, such cells showed a significantly enhanced migration rate in the chemotactic assay and decreased T-cell proliferation after lipopolysaccharide stimulation in mixed lymphocyte reactions. Cotransfected cells showed an increased ability to induce immune tolerance by upregulating T regulatory (Treg) cells and shifting the Th1/Th2 ratio. Cotransfection of Ad-CCR7 and RelB-siRNA endowed imDCs with resistance to apoptosis and cell death. CCR7 overexpression and RelB knockdown (KD) in imDCs improve skin-graft survival in a murine skin-transplantation model. CONCLUSION: Transfection with Ad-CCR7 and RelB KD in imDCs may be an effective approach inducing immune tolerance, thus being potentially valuable for inhibiting allograft rejection.

Astragaloside IV alleviates heart failure via activating PPARα to switch glycolysis to fatty acid ß-oxidation.

In heart failure (HF), energy metabolism pathway in cardiac muscle changes from fatty acid ß-oxidation to glycolysis. However, the exact mechanism is unknown. Sarcoendoplasmic reticulum Ca2+α ATPase (SERCA) expression is downregulated and mitochondrial function is reduced in HF, perhaps partly due to a substantially reduced energy supply for excitation-contraction coupling resulting from a lower fatty acid ß-oxidation rate. We investigated whether Astragaloside IV can activate peroxisome proliferator-activated receptor alpha (PPARα) to stimulate fatty acid ß-oxidation and increase cardiac energy production, improving mitochondrial function and the efficiency of SERCA in HF. In pressure overload-induced HF mice and isolated hypertrophic myocardial cells, fatty acid ß-oxidation and heart function were substantially strengthened following Astragaloside IV treatment, as demonstrated by the increased expression of PPARα and SERCA2a. In vitro, Astragaloside IV regulated energy metabolism by increasing ATP production and enhancing mitochondrial function, attributable to increased oxygen consumption and slightly increased mitochondrial Ca2+ uptake. In HF, Astragaloside IV switched glycolysis to fatty acid ß-oxidation, as confirmed by reduced anaerobic glycolysis and an increased oxygen consumption ratio. These results suggest that Astragaloside IV can stimulate fatty acid ß-oxidation and improve mitochondrial function, which may present a novel cardioprotective treatment that inhibits the progress of HF.

Astragaloside-IV Alleviates Heat-Induced Inflammation by Inhibiting Endoplasmic Reticulum Stress and Autophagy.

BACKGROUND: Thermal injury is the main cause of pulmonary disease in stroke after burn and can be life threatening. Heat-induced inflammation is an important factor that triggers a series of induces pathological changes. However, this mechanism underlying heat-induced inflammation in thermal inhalation injury remains unclear. Studies have revealed that astragaloside-IV (AS-IV), a natural compound extracted from Astragalus membranaceus, has protective effects in inflammatory diseases. Here, we investigated whether the protective effects of AS-IV occur because of the suppression of heat-induced endoplasmic reticulum (ER) stress and excessive autophagy Methods: AS-IV was administered to Wistar rats after thermal inhalation injury and 16HBE140-cells were treated with AS-IV. TNF-α, IL-6, and IL-8 levels were determined by ELISA and real-time PCR. ER stress and autophagy were determined by western blot. Autophagic flux was measured by recording the fluorescence emission of the fusion protein mRFP-GFP-LC3 by dynamic live-cell imaging. RESULTS: AS-IV had protective effects against heat-induced reactive oxygen species production and attenuated ER stress. AS IV alleviated heat-induced excessive autophagy in vitro and in vivo. Excessive autophagy was attenuated by the PERK inhibitor GSK2656157 and eIF2α siRNA, suggesting that heat stress-induced autophagy can activate the PERK-eIF2α pathway. Beclin 1 and Atg5 siRNAs inhibited the upregulation of the inflammatory cytokines TNF-α, IL-6, and IL-8 after heat exposure. CONCLUSIONS: Thus, AS-IV may attenuate inflammatory responses by disrupting the crosstalk between autophagy and the PERK-eIF2α pathway and may be an ideal agent for treating inflammatory pulmonary diseases.