Folate in the United States Population and its Association with Congestive Heart Failure.

Background: To investigate the relationship between red blood cell (RBC) folate and congestive heart failure (CHF). Methods: We extracted the concentrations of RBC folate and collated CHF information from the National Health and Nutrition Examination Survey (NHANES) survey (12820 individuals). Weighted univariate logistic regression, weighted multivariate logistic regression, and restrictive cubic spline (RCS) were used to assess the relationship between RBC folate concentrations and CHF. Results: The unadjusted model showed that the highest tertile group of RBC folate concentration was significantly associated with a higher risk of CHF compared to the lowest tertile group of RBC folate levels (odds ratio [OR] = 3.09; 95% confidence interval [CI], 2.14-4.46). Similar trends were seen in the multivariate-adjusted analysis (OR = 1.98; 95% CI: 1.27-3.09). The OR was > 1.0 when the predicted RBC folate exceeded 2757 nmol/L in the RCS model, indicating that the risk of CHF was low and relatively stable up to a predicted RBC folate level of 2757 nmol/L, but began to increase rapidly thereafter (p = 0.001). Conclusions: The risk of CHF may be increased either by high RBC folate concentrations (highest tertile of RBC folate or > 2637 nmol/L) or by folate deficiency. Considering the two sides of the association between RBC folate and CHF, there is a need for large-scale clinical research to better investigate if the association between RBC folate and CHF is a cause-effect relationship, what are the underlying pathophysiological basis, as well as to identify optimal dietary folate equivalent (DFE) and RBC folate concentration intervals.

Cardiac T1ρ Mapping Values Affected by Age and Sex in a Healthy Chinese Cohort.

BACKGROUND: To facilitate the clinical use of cardiac T1ρ, it is important to understand the impact of age and sex on T1ρ values of the myocardium. PURPOSE: To investigate the impact of age and gender on myocardial T1ρ values. STUDY TYPE: Cross-sectional. POPULATION: Two hundred ten healthy Han Chinese volunteers without cardiovascular risk factors (85 males, mean age 34.4 ± 12.5 years; 125 females, mean age 37.9 ± 14.8 years). FIELD STRENGTH/SEQUENCE: 1.5 T; T1ρ-prepared steady-state free precession (T1ρ mapping) sequence. ASSESSMENT: Basal, mid, and apical short-axis left ventricular T1ρ maps were acquired. T1ρ maps acquired with spin-lock frequencies of 5 and 400 Hz were subtracted to create a myocardial fibrosis index (mFI) map. T1ρ and mFI values across different age decades, sex, and slice locations were compared. STATISTICAL TESTS: Shapiro-Wilk test, Student's t test, Mann-Whitney U test, linear regression analysis, one-way analysis of variance and intraclass correlation coefficient. SIGNIFICANCE: P value <0.05. RESULTS: Women had significantly higher T1ρ and mFI values than men (50.3 ± 2.0 msec vs. 47.7 ± 2.4 msec and 4.7 ± 1.0 msec vs. 4.3 ± 1.1 msec, respectively). Additionally, in males and females combined, there was a significant positive but weak correlation between T1ρ values and age (r = 0.27), while no correlation was observed between the mFI values and age (P = 0.969). DATA CONCLUSION: We report potential reference values for cardiac T1ρ by sex, age distribution, and slice location in a Chinese population. T1ρ was significantly correlated with age and sex, while mFI was only associated with sex. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 1.

Cardiac magnetic resonance assessment of cardiac function across chronic kidney disease stages.

BACKGROUND: Cardiovascular disease prevalence remains high among chronic kidney disease (CKD) patients. Mechanisms and treatments to improve prognosis remain of paramount important and imaging biomarkers of left ventricular myocardial structure and function have better defined the phenotype of renal cardiomyopathy. The left atrial function and right heart remain are less well reported in CKD. This study used cardiac MRI to assess the interplay of left atrial and right ventricular function. METHODS: In a cross-sectional study, we examined 58 CKD patients (Group I: stages 2-3, n = 25; Group II: stages 4-5, n = 33). Additionally, 26 age-matched healthy controls were included. Comprehensive CMR protocols (1.5T) were employed, encompassing cine imaging, native T1 and T2 mapping, and tissue tracking strain analysis. LV, RV, and LA structure, function, and strain parameters were assessed. RESULTS: Compared to healthy controls, both groups I and II exhibited impaired RV and LA function. RVEDVi and RVESVi showed significant increases in both groups I and II (p < 0.001). All LV, RV, and LA strain parameters were reduced in the patient groups (all p < 0.001). In the univariate binary logistic regression, several parameters, including age, blood pressure, RV volumes and LV/RV strain were found to have a statistically significant association with CKD. In a multivariable model adjusted for other confounders, RV GLS and left atrial strain remained as independent significant predictors. CONCLUSIONS: RV size, LA strain and volume assessed by CMR serve as markers of RV and LA cardiac dysfunction in CKD patients with preserved LVEF. Greater attention should be given to RV and LA dysfunction for early identification of cardiac dysfunction in CKD patients.

P2X7 receptor inhibition prevents atrial fibrillation in rodent models of depression.

AIMS: Depression, the most prevalent psychiatric disorder, is associated with the occurrence and development of atrial fibrillation (AF). P2X7 receptor (P2X7R) activation participates in the development of depression, but little attention has been given to its role in AF. This study was to investigate the effects of P2X7R on AF in depression models. METHODS AND RESULTS: Lipopolysaccharide (LPS) and chronic unpredictable stress (CUS) were carried out to induce depression in rodents. Behavioural assessments, atrial electrophysiological parameters, electrocardiogram (ECG) parameters, western blot, and histology were performed. Atrial fibrillation inducibility was increased in both LPS- and CUS-induced depression, along with the up-regulation of P2X7R in atria. CUS facilitated atrial fibrosis. CUS reduced heart rate variability (HRV) and increased the expression of TH and GAP43, representing autonomic dysfunction. Down-regulation of Nav1.5, Cav1.2, Kv1.5, Kv4.3, Cx40, and Cx43 in CUS indicated the abnormalities in ion channels. In addition, the expression levels of TLR4, P65, P-P65, NLRP3, ASC, caspase-1, and IL-1ß were elevated in depression models. Pharmacological inhibitor (Brilliant Blue G, BBG) or genetic deficiency of P2X7R significantly mitigated depressive-like behaviours; ameliorated electrophysiological deterioration and autonomic dysfunction; improved ion channel expression and atrial fibrosis; and prevented atrial NLRP3 inflammasome activation in the pathophysiological process of AF in depression models. CONCLUSION: LPS or CUS induces AF and promotes P2X7R-dependent activation of NLRP3 inflammasome, whereas pharmacological P2X7R inhibition or P2X7R genetic deficiency prevents atrial remodelling without interrupting normal atrial physiological functions. Our results point to P2X7R as an important factor in the pathology of AF in depression.

The Experience in Lower Eyelid Blepharoplasty: Improvement of Surgical Procedures Based on Anatomical Basis.

OBJECTIVE: This study characterizes the appearance changes associated with aging of the human lower eyelid, grounded in its anatomical basis. Tailored approaches to lower eyelid bag are performed by these anatomical manifestations. METHODS: From January 2017 to January 2023, lower eyelid blepharoplasty was performed on 137 patients, aged 20 to 60 years. These patients were divided into 3 groups according to the periorbital aging appearance, primarily characterized by the presence and location of the "bag" bulge of the lower eyelid. We analyzed the corresponding changes in anatomical structures for each type, which included a weakened fibrous orbital support system, reduced muscle tone, and increased orbital fat. Patients were treated with tailored blepharoplasty techniques according to their classification. All patients in this study ranged in follow-up from 1 to 12 months. With patients' permit, photographs and clinical information were taken to evaluate the preoperative and postoperative outcome. RESULTS: Our study identified 3 morphologies of the lower eyelid. Type 1 presents a "bubble bag" in the medial and inferior aspect of the lower eyelid. Type 2 features a double convexity contour, with separate fat pad herniations demarcated by fibrous connective tissue. Type 3 exhibits a single convexity with a uniform herniation of fat pads across the entire lower eyelid. We have delineated the anatomical changes associated with each morphology type. With an approach grounded in "remodeling" and "recovery," the surgical treatment targets the fibrous support tissue to improve the outcomes of lower eyelid rejuvenation. No complications occurred. All postoperative results reached both surgeon's and patient's expectations. CONCLUSIONS: Surgeons must recognize the pivotal role of fibrous connective tissues-including the arcuate expansion, fascia of the inferior oblique muscle, and the orbicularis retaining ligament-and endeavor to preserve or reinforce these structures during surgical procedures. An anatomically based surgical approach would more effectively and safely to resist the facial aging process.

A Feasibility Study of HoloLens Ear Image Guidance for Ear Reconstruction.

BACKGROUND: An ideal 3D cartilage framework and accurate anatomical location are the most important factors to carry out a satisfactory reconstruction of the ear. To streamline this process, we developed an augmented reality assistance system, HoloLens Ear Image Guidance, which is based on computed tomography (CT) data, tailor-made navigation kits to guide reconstruction, data processing software, and HoloLens hardware. The objective of this study is to verify its feasibility in a clinical setting. METHODS: This study first validated our system in healthy controls and then extrapolated data to test on patients with microtia. First, three healthy volunteers were recruited, and reconstructive navigation kits were made using 3D printing. CT data were collected for the head and neck and imported into the HoloLens Ear Image Guidance Application to generate a personalized 3D virtual ear image. Volunteers then wore the navigation kits while researchers observed them through the HoloLens to check accuracy, track delay, and view the ear image guide.Ten patients with unilateral microtia were recruited and CT data were collected, and reconstructive navigation kits were made to assist with surgery. The procedure was monitored to record the surgeon's experience wearing the HoloLens, the patients' complications associated with wearing navigation kit, and to measure the symmetry between the reconstructed ear and the reference counterpart. RESULTS: In control patients, the deviation between the virtual image and the real ear was less than 2.4% (±0.22%); the tracking delay was less than 1.26 s (±0.09 s), the display effect was good, and surgeons did not report discomfort or dizziness while wearing the HoloLens. Volunteers did not report any pain from holding the navigation reference in their mouth during the test. Following validation, the HoloLens-assisted procedures were not associated with surgeon discomfort or dizziness. No complications were noted in patients including injury to the oral mucosa. Symmetry between the reconstructed ear and the contralateral ear was noted to be satisfactory in HoloLens-assisted surgery. CONCLUSIONS: The HoloLens Ear Image Guidance initially met clinical demands in registration accuracy, tracking speed, and subjective user experience, which can be used as the basis for continual software improvements and clinical application.

To Explore Ideas From the Altered Metabolites: The Metabolomics of Pathological Scar.

BACKGROUND: Pathological scars are dermal fibroproliferative disorders due to rapid inflammatory response after dermal injury. The altered metabolites could reflect pathophysiological changes directly. However, it has not cleared how the metabolites change scars. OBJECTIVE: To explore new ideas of pathological scars from the altered metabolites by using ultra-performance liquid chromatography coupled to tandem mass spectrometry and identifying the key genes. METHODS: Keloid (KS, n = 10), hypertrophic scar (HS, n = 10), and normal skin (NS, n = 10) were collected. Ultra-performance liquid chromatography coupled to tandem mass spectrometry was used to identify and characterize metabolites. Differential metabolites were analyzed by orthogonal partial least square discriminant analysis and Student t test. The key pathways were analyzed via Kyoto Encyclopedia of Genes and Genomes, and the related enzymes were verified by real-time Polymerase Chain Reaction, both in tissues and their dermal fibroblasts. RESULTS: Two hundred fourteen metabolites were detected in total, mostly were fatty acids and amino acids. In the KS and NS groups, 65 different metabolites were screened ( P < 0.05), and the polyunsaturated fatty acids (PUFAs) metabolism and butyric acid in keloid should be concerned. The messenger Ribonucleic Acid expression of fatty acid desaturase 1 and fatty acid desaturase 2, which are the key enzyme of PUFA metabolism, were lower in KS and keloid-derived fibroblasts, P < 0.05. In HS group, 17 metabolites were significantly different and branched chain amino acids degradation was the key pathway. Moreover, branched chain keto acid dehydrogenase E1 subunit alpha was lower expressed in HS and their fibroblasts compared with NS, P < 0.05. CONCLUSIONS: Polyunsaturated fatty acids and butyric acid may be associated with the generation of keloids. The pathogenesis of hypertrophic scars may be involved in branched chain amino acids degradation, which is worth paying attention to.

The role of altered fatty acid in pathological scars and their dermal fibroblasts.

PURPOSE: The proposed pathological mechanism for scar formation is controversial, and increased attention has been paid to the fatty acids (FAs) in the formation of pathological scars. Notably, FAs are known to be important in inflammation and mechanotransduction, which is closely related to scar formation. Therefore, it is necessary to clarify the roles of FA in scar formation. METHODS: Hypertrophic scar and keloid formed for more than a year and without other treatment, as well as normal skin samples were obtained from patients who underwent plastic surgery. Finally, keloids (n = 10), hypertrophic scars (n = 10), and normal skin samples (n = 10) were collected under informed consent. Primary dermal fibroblasts were isolated and cultured. The amount and variety of FAs were detected by lipid chromatography-mass spectrometry. Immunohistochemistry, real-time PCR, and western blotting were used to verify the expression of sterol regulatory element-binding protein-1 (SREBP1) and fatty acid synthase (FASN) in the samples and their fibroblasts. Student's t-test, ANOVA, and orthogonal partial least square discriminant analysis were performed for statistical analysis (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). RESULTS: Compared with full-thickness normal skin, there were 27 differential FAs in keloids and 15 differential FAs in hypertrophic scars (∗p < 0.05 and variable influence on projection >1.0). The expression of SREBP1 and FASN was lower in pathological scars both at mRNA and protein levels (all ∗p < 0.05). However, the mRNA levels of SREBP1 (∗∗∗p = 0.0002) and FASN (∗∗∗p = 0.0021) in keloid-derived fibroblasts were higher than that in normal skin fibroblasts (NFBs), while the expression in hypertrophic scar-derived fibroblasts was lower than that in NFBs (both ∗p < 0.05). Whereas there was no significant difference in FASN protein expression between keloid-derived fibroblasts and NFBs (p > 0.05). CONCLUSION: FAs involved in pathological scars are abnormally changed in scar formation. Thus, fatty acid-derived inflammation and de novo synthesis pathway of FA may play a key role in the formation of pathological scars.

Chyloid Fat Carried Adipose-Derived Mesenchymal Stem Cells Accelerate Wound Healing Via Promoting Angiogenesis.

ABSTRACT: Impaired wound healing is responsible for significant morbidity and mortality worldwide. It is necessary to find a stable, efficient, and safe method to promote soft tissue wound healing. Fat grafting has become increasingly popular in contouring procedures. However, more recently, there has been an emphasis on its regenerative potential. In this study, we established the wound healing model using nude mice. Hematoxylin and eosin and Masson stainings were performed to assess the effect of chyloid fat on the histology of wound healing. A laser Doppler perfusion imager was used to evaluate the blood perfusion of wounds. Immunohistochemistry was carried out to detect the expression of CD31 in wound tissues. The results suggested that after treatment with granule fat or chyloid fat, wound healing was accelerated and blood perfusion was promoted. In addition, granule fat or chyloid fat treatment promoted the angiogenesis of the wound. In addition, we evaluated the amount of adipose-derived mesenchymal stem cells in chyloid fat and granule fat. It was found that chyloid fat contained more adipose-derived mesenchymal stem cells than granule fat did. In conclusion, we proved that chyloid fat could significantly accelerate the wound healing process via promoting angiogenesis. The adipose-derived mesenchymal stem cell plays a critical role in this effect of chyloid fat.

RNA-binding protein SFPQ cooperates with HDAC1 to suppress CD40 transcription in pulmonary adventitial fibroblasts.

Pulmonary artery adventitial fibroblasts, the most abundant cellular constituent of adventitia, are often the first to be activated and reprogrammed to then influence the tone and structure of the vessel wall in pulmonary arterial hypertension (PAH). Our previous study found that interruption of CD40 ligand (CD40L)-CD40 signaling improves the efficacy of transplanted endothelial progenitor cells in monocrotaline induced-PAH. However, whether CD40L-CD40 signaling is involved in the activation of adventitial fibroblasts in PAH and whether Drosophila behavior human splicing (DBHS) protein family members have any roles during adventitial fibroblasts activation are completely unclear. Here, we show that soluble CD40L (sCD40L) stimulation progressively increases pro-inflammatory activity, proliferation, and migration of pulmonary adventitial fibroblasts. Besides, sCD40L stimulation decreases splicing factor proline- and glutamine-rich protein (SFPQ) protein (one member of DBHS protein family) expression, while SFPQ overexpression suppresses sCD40L stimulation-induced proliferation and migration of pulmonary adventitial fibroblasts by repressing CD40 transcription. Moreover, ChIP assays found that sCD40L stimulation promotes histone H3 tri-methylation at lysine 4 (H3K4me3), H3K36me3, and H3K27 acetylation (H3K27ac) modifications on CD40 promoter region in pulmonary adventitial fibroblasts, while SFPQ overexpression decreases H3K36me3 modification and increases H3K36ac on CD40 promoter region by interacting with histone deacetylase-1 (HDAC1) to inhibit CD40 transcription. This in-depth study shows that CD40L-CD40 signaling promotes activation of pulmonary adventitial fibroblasts by increasing proliferation, migration, and pro-inflammatory activity of adventitial fibroblasts, and SFPQ could inhibit CD40 transcription though switching H3K36me3 to H3K36ac modifications on its promoter by interacting with HDAC1. This study, first, uncovers the roles of SFPQ on CD40L-CD40 signaling-mediated activation of pulmonary adventitial fibroblasts.

Yin Yang-1 suppresses CD40 ligand-CD40 signaling-mediated anti-inflammatory cytokine interleukin-10 expression in pulmonary adventitial fibroblasts by promoting histone H3 tri-methylation at lysine 27 modification on interleukin-10 promoter.

During the pathogenesis of early pulmonary arterial hypertension (PAH), pulmonary arterial adventitial fibroblast act as an initiator and mediator of inflammatory processes that predispose vessel walls to excessive vasoconstriction and pathogenic vascular remodeling. Emerging studies report that Yin Yang-1 (YY-1) plays important roles in inflammatory response and vascular injury. Our recent study finds that activation of CD40 ligand (CD40L)-CD40 signaling promotes pro-inflammatory phenotype of pulmonary adventitial fibroblasts. However, whether YY-1 is involved in CD40L-CD40 signaling-triggered inflammatory response in pulmonary adventitial fibroblasts and its underlying mechanism is still unclear. Here, we show that soluble CD40L (sCD40L) stimulation promotes YY-1 protein expression and suppresses anti-inflammatory cytokine, interleukin 10 (IL-10) expression in pulmonary adventitial fibroblasts, while YY-1 knockdown prevents sCD40L-mediated reduction of IL-10 expression via enhancing IL-10 gene transactivation. Further, we find that sCD40L stimulation significantly increases histone H3 tri-methylation at lysine 27 (H3K27me3) modification on IL-10 promoter in pulmonary adventitial fibroblasts, and YY-1 knockdown prevents the effect of sCD40L on IL-10 promoter by reducing the interaction with enhancer of zeste homolog 2 (EZH2), a histone methyltransferase, binding to IL-10 promoter. Moreover, we find that sCD40L stimulation promotes YY-1 protein, but not messenger RNA (mRNA) expression, via decreasing N6-methyladenosine methylation on YY-1 mRNA to suppress YTHDF2-medicated mRNA decay. Overall, this in-depth study shows that the activation of CD40L-CD40 signaling upregulates YY-1 protein expression in pulmonary adventitial fibroblasts, which results in increasing YY-1 and EZH2 binding to the IL-10 promoter region to enhance H3K27me3 modification, eventually leading to suppression of IL-10 transactivation. This study first uncovers the roles of YY-1 on CD40L-CD40 signaling-triggered inflammatory response in pulmonary adventitial fibroblasts.

Mass Spectrometric Characterization of Metabolites in Ear Cartilage: Congenital Microtia and Normal Auricle.

OBJECTIVE: The aims of the study were to perform mass spectrometric characterization of metabolites in microtic and healthy ear auricular cartilage tissue, to screen the differential metabolites and pathways in these tissues, and to find a connection between the changes in the metabolic pathways and the biochemical properties of the cartilage tissue. METHODS: According to the inclusion criteria, patients with simple microtia admitted to the hospital between June 2017 and January 2018 were selected upon admission. During ear reconstruction surgery, residual auricle cartilage tissues of the patients were harvested as the case group (18 cases), and normal auricle cartilage tissues (18 cases) were taken as the control group. The mass spectrometry technique gas chromatography time-of-flight mass spectrometry and the Xplore platform were used to identify and characterize the metabolites in the ear cartilage samples. Then, differential metabolites and key pathways were identified and analyzed. RESULTS: In total, 277 metabolites were detected, but only 132 metabolites were annotated in the JiaLib (one of the largest metabolomics libraries in the world). Of those, 14 differential metabolites and 3 metabolic pathways were identified between microtia and healthy ear cartilage, including the pathways of arginine metabolism, taurine metabolism, and pantothenate and CoA metabolism, P < 0.05. CONCLUSIONS: Arginine, taurine, and L-cysteine may have an association with the development of microtia ear cartilage, and arginine succinate synthase and argininosuccinate lyase may be the key enzyme in microtia. This new direction on microtia can help us understand the pathogenesis of microtia and propose some new ideas for its etiology.

The Regenerated Tissue at the Donor Site After Costal Cartilage Harvest for Auricular Reconstruction.

OBJECTIVE: To observe and summarize the nature of the regenerative tissue at the donor site after harvesting costal cartilage for auricular reconstruction and to explore the contribution of the perichondrium to the regeneration of costal cartilage in the clinic. METHODS: From January 2016 to June 2017, 23 patients with microtia who were performed chest computed tomography (CT) after costal cartilage harvest for ear reconstruction were reviewed. And they had the surgery for at least 6 months. Of 23 patients, 17 patients were males and 6 were females; these patients were aged 7 to 43 years (mean age, 15.2 years). The authors divided the patients into 2 groups according to whether the perichondrium was retained or not. Group 1 was patients with intact perichondrium, total 20. Group 2 was patients with damaged perichondrium, total 3. Every patients' regenerative tissue CT value at the donor-site region of costal cartilage was measured and recorded. In addition, 2 regenerated tissue samples for examined histologic evaluation by hematoxylin and eosin stain were collected. RESULTS: Of 23 patients, regenerated tissue with high CT value (above 100 Hounsfield unit [Hu]) was observed in 19 (82.61%) patients from group 1. And the direction of the regenerated tissue is roughly similar to that of the resected cartilage in the early surgery. Of 4 patients (1 from group and 3 from group 2), nothing on the donor site was found. From histologic evaluation, fibrocalcific tissue was seen, and cartilage cells were not seen in 2 patients with high CT value. CONCLUSION: Clinical observation presented that regenerative tissue at the donor site after harvesting costal cartilage, leaving the subjacent perichondrium completely intact, was mostly fibrocalcific tissue rather than cartilage tissue. The authors suspect that the perichondrium itself may not have regenerative power, but as an envelope for regeneration, perichondrium has a role.

The Role of RhoA in Neovascular-Related Functions of Endothelial Progenitor Cells Induced by Angiotensinâ ¡.

BACKGROUND/AIMS: Interference with endothelial progenitor cell (EPC) neovascularization is a novel therapeutic target for neovascular-related diseases. Angiotensin Ⅱ (Ang Ⅱ) was found to enhance new vessel formation and aggravated neovascular-related diseases. In this study, we investigated the effects of Ang Ⅱ on EPC neovascular-related functions and explored the underlying mechanisms. METHODS: EPCs were cultured from bone marrow derived mononuclear cells. The effects of Ang Ⅱ on EPC proliferation, adhesion, migration, and in vitro tube formation were investigated using the MTT assay, adhesion assay, transwell chamber assay, and in vitro tube formation assay respectively. The underlying mechanisms were explored using Western blotting assay. RESULTS: EPC adhesion, migration and in vitro tube formation were promoted by Ang Ⅱ, and the effects were reversed by RhoA/Rho-associated kinases (ROCK) signaling pathway inhibitors including C3 exoenzyme, GGTI-286 and Y-27632. The active form of RhoA was up-regulated by Ang Ⅱ and this effect was abolished by C3 exoenzyme. Moreover, RhoA silencing resulted in a notable inhibition of EPC adhesion, migration and in vitro tube formation, suggesting that RhoA activation played a pivotal role in Ang Ⅱ angiogenic effect. The results also demonstrated that phosphorylation of p38 mitogen-activated protein kinase (MAPK) and c-Jun-NH2 kinase was elevated by Ang Ⅱ and attenuated by C3 exoenzyme, GGTI-286 and Y-27632. The enhancing effects of Ang Ⅱ on EPC adhesion, migration and in vitro vasculogenesis were reversed by p38 inhibitor SB202190 and JNK inhibitor SP600125. CONCLUSION: Ang Ⅱ may enhance EPC neovascular-related functions through activating RhoA/ ROCK and MAPK signaling pathway.

Cryptotanshinone inhibits VEGF-induced angiogenesis by targeting the VEGFR2 signaling pathway.

OBJECTIVE: Anti-angiogenesis has been proposed as an important strategy for angiogenesis-related diseases. Cryptotanshinone (CPT), an active component of Salvia miltiorrhiza, may be a potential inhibitor of angiogenesis. However, the molecular mechanisms underlying its anti-angiogenic activities remain poorly understood. This study is to investigate the effects of CPT on VEGF-induced angiogenesis and VEGFR2 signaling pathway in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were treated with different concentration of CPT (5-20µmol/L) and the viability, endothelial cell migration, invasion, and tubular-like structure formation of HUVECs were detected by MTT, wound-healing migration, Transwell invasion and Matrigel tube formation assays, respectively. To assess the effect of CPT on VEGFR2 signaling pathway, VEGF-induced phosphorylation of VEGFR2 and its downstream molecules, including ERK1/2, p90RSK, Src and FAK were analyzed by Western blot. RESULTS: CPT significantly suppressed VEGF-induced cells proliferation, migration, invasion, and tubular-like structure formation in HUVECs in a dose- and time-dependent manner. Western blot results revealed that CPT significantly suppressed VEGF-induced phosphorylation of VEGFR2 and its key downstream protein kinases, including p-ERK1/2, p-p90RSK, pY416-Src and pY576/577-FAK, which are responsible for endothelial cell migration, proliferation, and survival. CONCLUSION: Our study suggested that CPT potently inhibits VEGF-induced angiogenesis by suppressing VEGFR2 activation and its downstream Src/FAK and ERK1/2 signaling pathways in HUVECs, highlighting the therapeutic potential for the treatment of angiogenesis-related diseases.

Tanshinone II A Attenuates TNF-α-Induced Expression of VCAM-1 and ICAM-1 in Endothelial Progenitor Cells by Blocking Activation of NF-κB.

BACKGROUND/AIMS: Tanshinone IIA (Tan IIA) is effective in the treatment of inflammation and atherosclerosis. The adhesion of inflammatory cells to vascular endothelium plays important role in atherogenic processes. This study examined the effects of Tan IIA on expression of adhesion molecules in tumor necrosis factor-α (TNF-α)-induced endothelial progenitor cells (EPCs). METHODS: EPCs were pretreated with Tan IIA and stimulated with TNF-α. Mononuclear cell (MNC) adhesion assay was performed to assess the effects of Tan IIA on TNF-α-induced MNC adhesion. Expression of vascular cell adhesion molecule-1 (VCAM-1)/intracellular adhesion molecule-1 (ICAM-1) and activation of Nuclear factor κB (NF-κB) signaling pathway were measured. RESULTS: The results showed that the adhesion of MNCs to TNF-α-induced EPCs and expression of VCAM-1/ICAM-1 in EPCs were promoted by TNF-α, which were reduced by Tan IIA. TNF-α increased the amount of phosphorylation of NF-κB, IκB-α and IKKα/ß in cytosolic fractions and NF-κB p65 in nucleus, while Tan IIA reduced its amount. CONCLUSION: This study demonstrated a novel mechanism for the anti-inflammatory/anti-atherosclerotic activity of Tan IIA, which may involve down-regulation of VCAM-1 and ICAM-1 through partial blockage of TNF-α-induced NF-κB activation and IκB-α phosphorylation by the inhibition of IKKα/ß pathway in EPCs.

Interruption of CD40 Pathway Improves Efficacy of Transplanted Endothelial Progenitor Cells in Monocrotaline Induced Pulmonary Arterial Hypertension.

BACKGROUND/AIMS: Transplantation of endothelial progenitor cells (EPCs) plays a therapeutic role in pulmonary arterial hypertension (PAH). Meanwhile, recruitment of progenitors has potential inflammatory effects and exaggerates vascular injury. CD40 pathway is identified as a major player in vascular inflammatory events. In this study, we investigated the role of CD40 pathway in regulating early outgrowth EPC functions, and searched for improvements in PAH cell therapy. METHODS: EPCs were isolated from rat bone marrow and cultured for 7 days. After treatment with soluble CD40 ligand (sCD40L) for 24 hours, EPC migration, adhesion, proliferation, paracrine and vasculogenesis functions were tested. Rat PAH model was founded by subcutaneous injection of monocrotaline (MCT). Control EPCs or lentivirus vectors (Lv)-shRNA-CD40 EPCs were infused via tail vein at day 7, 14, and 21 after MCT injection. Therapeutic effects were evaluated at day 28. RESULTS: sCD40L dose-dependently impaired EPC migration, adhesion, proliferation, and vasculogenesis functions. However, paracrine effects of soluble intercellular adhesion molecule-1, vascular endothelial growth factor and interleukin-6 were dose-dependently improved by sCD40L. Control EPC-derived conditioned medium protected endothelial cell in vitro vasculogenesis, while sCD40L-pretreated ones showed detrimental effects. After MCT injection, sCD40L levels in rat serum increased gradually. Other than in vitro results, benefits of both two EPC treatments were obvious, even taken at day 21. Benefits of control EPCs wore off over time, but those of Lv-shRNA-CD40 EPCs were more effective and enduring, as characterized by both ameliorated rat hemodynamic and reversed vascular remodeling. Furthermore, Lv-shRNA-CD40 EPCs integrated into endothelium better, rather than into adventitia and media. CONCLUSION: sCD40L impaired protective effects of EPCs. Traditional EPC treatments were limited in PAH, while interruption of CD40 pathway of transplanted cells could apparently improve the therapeutic efficacy.

Activation of extracellular signal-regulated kinase 1/2 and Sp1 may contribute to the expression of tissue inhibitor of metalloproteinases-1 induced by transforming growth factor-ß1 in human pulmonary arterial smooth muscle cells.

BACKGROUND AIMS: Tissue inhibitor of metalloproteinases-1 (TIMP-1) plays an important role in the development of pulmonary arterial hypertension. However, the molecular regulatory mechanisms of TIMP-1 in the pulmonary arteries are not fully understood, especially in human pulmonary arterial smooth muscle cells (HPASMCs). We investigated the signaling pathway involved in the regulation of TIMP-1 in HPASMCs induced by transforming growth factor (TGF)-ß1. METHODS: Cultured HPASMCs were incubated with different concentrations of TGF-ß1 (0-40 ng/mL) for 24 h or with 10 ng/mL TGF-ß1 for different times (1-48 h). RESULTS: Western blot, real-time polymerase chain reaction and enzyme-linked immunosorbent assay analyses showed that TGF-ß1 enhanced the expression and secretion of TIMP-1 in a time-dependent and dose-dependent fashion. TGF-ß1 could phosphorylate two of the three mitogen-activated protein kinases-extracellular signal-regulated kinase 1/2 (ERK1/2) and p38, but not c-Jun NH2-terminal kinase. Of these kinases, only the inhibition of ERK1/2 by U0126, which was a specific inhibitor of mitogen-activated protein kinase/ERK1/2, effectively blocked the TGF-ß1-induced expression of TIMP-1. Mithramycin, an inhibitor of Sp1 transcription factor, also significantly inhibited the expression of TIMP-1. Additionally, electrophoretic mobility shift assay showed that TGF-ß1 could up-regulate the DNA-binding activity of Sp1 and that U0126 and mithramycin could effectively inhibit these events. CONCLUSIONS: TGF-ß1 could stimulate the expression and secretion of TIMP-1 in HPASMCs in a time-dependent and dose-dependent fashion, and ERK1/2 and Sp1 signaling pathways might be involved in these activities.

Discrimination of Left Atrial Strain Patterns in Hypertensive Heart Disease and Hypertrophic Cardiomyopathy: a Cardiac Magnetic Resonance Feature.

To assess left atrial (LA) strain parameters using cardiovascular magnetic resonance imaging feature tracking (cardiac MRI-FT) for differentiating hypertensive heart disease (HHD) from hypertrophic cardiomyopathy (HCM), which are two left ventricular hypertrophic diseases that could present with similar morphologies in early stage but differ in clinical symptoms and treatment strategies. 45 patients with HHD, 85 patients with HCM (non-obstructive hypertrophic cardiomyopathy [HNCM, n = 45] and obstructive hypertrophic cardiomyopathy [HOCM, n = 40]) and 30 healthy controls (HC) were retrospectively included. LA volumes, strain, and strain rate were determined by manually contouring on the two- and four-chamber views of the CMR-FT module using CVI 42 software. LA volume parameters including LA maximum, precontraction, and minimum volume index, and total, passive, and active emptying fractions were obtained using the biplane methods. The LA strain parameters, including total strain (εs), passive strain (εe), active strain (εa), peak positive strain rate (SRs), early peak negative strain rate (SRe), and late peak negative strain rate (SRa), were obtained from the LA strain curve. The LA strain and LA strain rate were impaired in both HHD group and HCM group, and they were the most severely impaired in the HOCM group. εs (AUC = 0.691, P = 0.006; the best cutoff value, 25.1%), εa (AUC = 0.654, P = 0.027; the best cutoff value, 10.5%), SRs (AUC = 0.710, P = 0.003; the best cutoff value, 0.81 1/s) and SRa (AUC = 0.667, P = 0.016; the best cutoff value, -1.30 1/s) showed significant differences in the identification between HHD and HNCM. All LA strain parameters were different in the identification between HHD and HOCM (all P < 0.05).LA strain parameters can be helpful for differentiating HHD from HCM, providing valuable insights for diagnosis.

Changes in subclinical cardiac abnormalities 1 Year after recovering from COVID-19 in patients without clinical cardiac findings.

Aim: To evaluate the subclinical cardiac involvement in COVID-19 patients without clinical cardiac evidence using cardiac MR imaging. Material and methods: Participants recovered from COVID-19 without cardiac symptoms and no cardiovascular medical history were enrolled in a prospective cohort study. They underwent baseline cardiac MR and follow-up cardiac MR > 300 days after discharge (n = 20). The study also included healthy controls (n = 20). Extracellular volume fraction (ECV), native T1, and 2D strain data were assessed and compared. Results: The ECV values of participants at baseline [30.0% (28.3%-32.5%)] and at follow-up [31.0% (28.0%-32.8%)] were increased compared to the healthy control group [27.0% (25.3%-28.0%)] (both p < 0.001). However, the ECV increase from baseline cardiac MR to follow-up cardiac MR was not significant (p = 0.378). There was a statistically significant difference in global native T1 between baseline [1140 (1108.3-1192.0) ms] and follow-up [1176.0 (1113.0-1206.3) ms] (p = 0.016). However, no native T1 difference was found between the healthy controls [1160.7 (1119.6-1195.4) ms] and the baseline (p = 0.394) or follow-up group (p = 0.168). The global T2 was 41(40-42) ms at follow-up which was within the normal range. In addition, We found a recovery in 2D GLS among COVID-19 participants between baseline and follow-up [-12.4(-11.7 to -14.3)% vs. -17.2(-16.2 to -18.3)%; p＜0.001]. Conclusion: Using cardiac MR myocardial tissue and strain imaging parameters, 35% of people without cardiac symptoms or clinical evidence of myocardial injury still had subclinical myocardial tissue characteristic abnormalities at 300 days, but 2D GLS had recovered.