EMX2 inhibits clear cell renal cell carcinoma progress via modulating Akt/FOXO3a pathway.

Empty spiracles homeobox 2 (EMX2) is initially identified as a key transcription factor that plays an essential role in the regulation of neuronal development and some brain disorders. Recently, several studies emphasized that EMX2 could as a tumor suppressor, but its role in human clear cell renal cell carcinoma (ccRCC) remains unclear. In the present study, we investigated the role and underlying mechanism of EMX2 in the regulation of ccRCC progress. Our results demonstrated that EMX2 expression was markedly decreased in ccRCC tissues and cell lines, and low EMX2 expression predicted the poor prognosis of ccRCC patients. In addition, forced expression of EMX2 significantly inhibited the cell growth, migration, and invasion in vitro, as well as ccRCC tumor growth in nude mice, via, at least in part, regulating Akt/FOXO3a pathway. In detail, EMX2 could attenuate the phosphorylation levels of Akt and FOXO3a, and increase FOXO3a expression without affecting total Akt expression in vivo and in vitro. Meanwhile, shRNA-mediated knockdown of FOXO3a expression could obviously attenuate the effects of EMX2 on cell growth, migration, invasion, and tumor growth. Furthermore, EMX2 could significantly attenuate the interaction between Akt and FOXO3a. Taken together, our results demonstrated that EMX2 could inhibit ccRCC progress through, at least in part, modulating Akt/FOXO3a signaling pathway, thus representing a novel role and underlying mechanism of EMX2 in the regulation of ccRCC progress.

Light-induced circadian rhythm disorder leads to microvascular dysfunction via up-regulating NETs.

Circadian rhythm is a physical, mental, and behavioral pattern over the course of 24-hour cycle, and its disturbance is associated with increased risk of cardiovascular diseases. Microvascular dysfunction serves as an important cause of cardiovascular disease, but the relationship between rhythm disturbances and microcirculation remains elusive. Herein, we constructed the mice model of circadian rhythm disturbance and investigated the alterations of microvascular conditions. It was revealed that coronary microcirculatory function and cardiac diastolic function were significantly reduced, along with endothelium-dependent diastolic function of microvessels remarkably impaired in the rhythm-disordered group of mice compared to the control group. Notably, rhythm disturbance led to a significant upregulation of neutrophil extracellular traps (NETs) levels in mice, which cause endothelial dysfunction by inhibiting microvascular endothelial cell activity and migration capacity as well as inducing apoptosis. Additionally, intraperitoneal injection of Cl-amidine suppressed the production of NETs, which further improved coronary microcirculatory function and endothelium-dependent diastolic function. In conclusion, this study demonstrated that circadian rhythm disorders could induce the development of coronary microvascular dysfunction (CMD) through the up-regulation of NETs, providing a potential therapeutic direction for the treatment of CMD.

Base-Controlled Divergent Synthesis of Fluorine-Containing Benzo[4,5]imidazo[2,1-b][1,3]thiazines from 2-Mercaptobenzimidazoles and ß-CF3-1,3-Enynes.

A base-controlled divergent cyclization between 2-mercaptobenzimidazoles and ß-CF3-1,3-enynes providing either trifluoromethylated or fluorinated benzo[4,5]imidazo[2,1-b][1,3]thiazines has been developed. The ß-CF3-1,3-enyne, as a three-carbon synthon, underwent a 1,8-diazabicyclo[5.4.0] undec-7-ene (DBU)-catalyzed tandem hydroamination/intramolecular hydrothiolation to give CF3-substituted 3,4-dihydro-2H-benzo[4,5]imidazo[2,1-b][1,3]thiazine, whereas reaction with KOH afforded fluorinated 4H-benzo[4,5]imidazo[2,1-b][1,3]thiazine exclusively. In addition, the synthetic utility of this methodology was showcased through a variety of downstream derivatizations.

Gualou Xiebai Decoction ameliorates increased Caco-2 monolayer permeability induced by bile acids via tight junction regulation, oxidative stress suppression and apoptosis reduction.

Gualou Xiebai Decoction (GXD), a classic prescription, is widely used to dealing with inflammatory diseases in China for thousands of years. Abnormal metabolic state of bile acids (BAs) is confirmed to cause intestinal epithelial barrier dysfunction. In preliminary work, we observed that GXD could decrease intestinal permeability in hyperlipidemia mice. The present study aimed to explore the protective effect of GXD on intestinal mucosa in vitro. Caco-2 cell monolayer permeability among different groups was determined by measuring the concentrations of FITC-dextran in the lower compartments and transepithelial electrical resistance (TEER). Meanwhile, mRNA and protein expressions of tight junctions (TJs) were investigated. Generation of intracellular reactive oxygen species (ROS) and the ratio of cell apoptosis induced by BAs were assessed by fluorescence probe and flow cytometry. GXD was shown to keep the cell monolayer in low permeable status, increase TEER and mRNA and protein expressions of occludin (Ocln) and zonula occluden 2 (ZO2) remarkably in cells challenged with cholic acid (CA), deoxycholic acid (DCA) and glycocholic acid (GCA). However, no significant effects were uncovered against the pathological effects of taurocholic acid (TCA). Meanwhile, generation of ROS and increased levels of apoptotic cells caused by CA, DCA and GCA were dramatically decreased by GXD, which were not observed on TCA. GXD could significantly attenuate intestinal barrier dysfunction induced by BAs via TJs regulation, oxidative stress suppression and cell apoptosis decrease, but such effects and behind mechanisms differed among different kinds of BAs.

N-Trifluoropropylation of Azoles through N-Vinylation and Sequential Hydrogenation.

Installing a fluoroalkyl group onto the nitrogen atom of azoles represents a potential strategy for lead optimization in medicinal chemistry. Herein, we describe a method for the N-trifluoropropylation of azoles. This process is accomplished using a combination of regioselective N-vinylation and sequential hydrogenation. The two-step sequence is applicable to a diverse set of azoles and tolerates a wide range of functionalities. In addition, we showcase its practicability and utility through the gram-scale synthesis and the late-stage modification of a complex molecule.

Metabolomics reveal dynamic changes in eicosanoid profile in patients with ST-elevation myocardial infarction after percutaneous coronary intervention.

Eicosanoids play important roles in the cardiovascular system. The metabolic disorders involving some eicosanoids in the pathophysiologic process include myocardial infarction and myocardial ischaemia-reperfusion injury. Percutaneous coronary intervention (PCI) is often the first-choice strategy for acute ST-segment elevation myocardial infarction (STEMI) according to current guidelines. This study aimed to investigate the dynamic eicosanoid metabolic profile in STEMI patients after PCI. The eicosanoid profiles in plasma of 20 patients at seven times (30 minutes before surgery; 6, 12, 24, and 72 hours after surgery; 1 day before discharge; and 28 days after surgery) were studied by using metabolomics. Levels of PGE2, PGD2, and TXA2 were decreased significantly and EETs contents were increased significantly at 6 hours after PCI. EETs were hydrolysed to DHETs within a short time after surgery (12-72 hours). 20-HETE content was significantly increased. In samples taken at the time of discharge and at follow-up after discharge, LTB4 level continued to increase. This work suggests that change in content of some functional eicosanoids may be involved in cardiac injury and repair after PCI in a synergistic manner.

Pre-treatment with compound Danshen dripping pills prevents lipid infusion-induced microvascular dysfunction in mice.

CONTEXT: Recent studies have shown compound Danshen dripping pills (CDDP) could improve microcirculation in ischemic/reperfusion injury and other microvascular disorders. The mechanism for CDDP's role in microcirculation is not clear. OBJECTIVE: To explore the protective effects of CDDP on microvascular dysfunction. MATERIALS AND METHODS: C57BL/6 male mice (6-8 weeks) were randomized into control, model and CDDP groups (n = 10), which were treated with normal saline or CDDP (105.30 mg/kg), respectively. Then, lipid emulsion and heparin were infused via mice jugular vein to establish systemic microvascular dysfunction model. Coronary flow reserve (CFR) and leukocytes adhesion on microvascular wall were measured. Relative CD11b and CD62L expression levels on neutrophils were measured by flow cytometric analysis. Expression level of forkhead box transcription factor O1 (FOXO1) mRNA was identified by real-time PCR. RESULTS: Lipid infusion significantly attenuated the CFR (1.84 ± 0.14 vs. 2.65 ± 0.02) and increased the number of leukocytes adherent to microvascular wall in cremaster (4067.00 ± 581.20 cells/mm2 vs. 10.67 ± 4.81 cells/mm2). The expression level of CD11b and FOXO1 in neutrophils was also up-regulated by lipid infusion. Pre-treatment with CDDP significantly improved CFR (2.57 ± 0.29 vs. 1.84 ± 0.14), decreased the number of leukocytes adherent to microvascular wall (2500.00 ± 288.70 cells/mm2 vs. 4067.00 ± 581.20 cells/mm2) and down-regulated CD11b and FOXO1 expression. Discussion and conclusions: Pre-treatment with CDDP could prevent lipid infusion-induced systemic microvascular disorder including coronary and peripheral microvascular dysfunction. Down-regulated FOXO1 and decreased leukocyte adhesion might play an important role in the mechanisms of CDDP's efficacy.

Cutaneous microvascular function in patients with obstructive or non-obstructive coronary artery disease evaluated by laser speckle contrast imaging.

OBJECTIVE: This study aimed to compare cutaneous microvascular function in coronary artery disease (CAD) patients including obstructive CAD (ObCAD) (n = 133) and non-obstructive CAD (NObCAD) (n = 129) with that of age and gender matched healthy controls (n = 83) using laser speckle contrast imaging (LSCI) coupled with post-occlusive reactive hyperemia (PORH). METHODS: LSCI system was used to assess subjects' cutaneous blood flow at rest and during PORH. CAD patients were divided into ObCAD and NObCAD group based on coronary angiography results. RESULTS: Microvascular reactivity induced by PORH was significantly reduced in NObCAD group in comparison with control or ObCAD group (p < 0.05). Although, the PORH responses of ObCAD patients were also attenuated compared to controls, they did not reach statistical significance (p > 0.05). CONCLUSION: NObCAD patients are more likely to develop cutaneous microvascular dysfunction than ObCAD patients, which may reflect the difference in the underlying mechanisms of myocardial ischemia.

Shexiang Tongxin Dropping Pill Improves Peripheral Microvascular Blood Flow via Cystathionine-γ-Lyase.

BACKGROUND To explore the protective effects of Shexiang Tongxin Dropping Pill (STP) in improving peripheral microvascular dysfunction in mice and to explore the involved mechanism. MATERIAL AND METHODS A peripheral microvascular dysfunction model was established by combined myocardial infarction (MI) and lipopolysaccharide (LPS) injection in mice. Then, the mice were randomized into a model group (n=10) or an STP group (n=10), which were treated with normal saline and STP, respectively. The cremaster muscle microvascular blood flow velocity and numbers of leukocytes adherent to the venular wall were evaluated before and after drug intervention. We assessed the expression of adhesion molecule CD11b and related transcript factor FOXO1 in leukocytes, cystathionine-γ-lyase (CSE) mRNA expression in the cremaster muscle, and mitochondrial DNA copy numbers. RESULTS Compared with those of control mice, the cremaster microvascular blood flow velocity, cremaster CSE expression, and mitochondrial DNA copy number in mice from the model group were significantly lower and leukocyte adhesion and CD11b and FOXO1 expression were significantly higher. Intervention with STP could significantly increase the cremaster microvascular flow velocity (0.480±0.010 mm/s vs. 0.075±0.005 mm/s), mRNA expression of cremaster CSE, and mitochondrial DNA copy number, but it inhibited leukocyte adhesion and decreased leukocyte CD11b and FOXO1 expression. CONCLUSIONS STP significantly improved peripheral microcirculation, in which increased CSE expression might be the underlying mechanism.

Circadian locomotor output cycles kaput accelerates atherosclerotic plaque formation by upregulating plasminogen activator inhibitor-1 expression.

To explore the association between clock circadian regulator circadian locomotor output cycles kaput gene (CLOCK) and the forming of atherosclerotic plaques and its underlying mechanisms, mouse aortic endothelial cells (MAECs) and atherosclerosis (AS) mouse model were recruited for our study. The apoE gene knockout mouse was used as the model of AS and we accelerated the formation of unstable plaques through the combination of carotid artery ligation and high-fat (HF) diet administration (0.2% cholesterol, 20% fat). The mRNA and protein expressions of CLOCK in peripheral blood monouclear cells of acute coronary syndrome (ACS) patients or mouse AS model were detected by qPCR, western blot analysis and immunohistochemical staining. The number of adherent cells and atherosclerotic plaques was counted to assess the effects of CLOCK on the progression of ACS, and adherence-associated genes, such as vascular cell adhesion molecule (VCAM)-1, C-C motif chemokine ligand 2 (CCL-2), and CCL-5. The results showed that CLOCK expression was significantly increased in both ACS patients and AS mouse model. The levels of CLOCK, leukemia inhibitory factor (LIF), intercellular adhesion molecule 1 (ICAM-1), perilipin 2 (ADFP), nuclear factor kappa B (NF-κB), and plasminogen activator inhibitor-1 (PAI-1), as well as the number of atherosclerotic plaques were elevated in the AS mouse model, as compared with the control group. Chromatin immunoprecipitation assay showed that CLOCK bound directly to the promoter of PAI-1 gene and CLOCK could positively regulate the expressions of LIF, ICAM-1, ADFP, NF-κB, and PAI-1. Reduction of CLOCK expression would decrease the expressions of VCAM-1, CCL-2, and CCL-5, and the number of adherent cells and atherosclerotic plaques, but these effects were neutralized when PAI-1 was simultaneously overexpressed in either mouse model or MAECs. Our results demonstrate that CLOCK overexpression triggers the formation of atherosclerotic plaques by directly upregulating PAI-1 expression.

Obstructive sleep apnea combined dyslipidemia render additive effect on increasing atherosclerotic cardiovascular diseases prevalence.

BACKGROUND: Current study was designed to investigate the effects of obstructive sleep apnea (OSA) combined dyslipidemia on the prevalence of atherosclerotic cardiovascular diseases (ASCVD). METHODS: This was a cross-sectional study and subjects with documented dyslipidemia and without previous diagnosis of OSA were enrolled. Polysomnography was applied to evaluate apnea-hypopnea index (AHI). Based on AHI value, subjects were classified into four groups: without OSA, mild, moderate and severe OSA groups. Clinical characteristics and laboratory examination data were recorded. Relationship between AHI event and lipid profiles was analyzed, and logistic regression analysis was used to evaluate the effects of OSA combined dyslipidemia on ASCVD prevalence. RESULTS: Totally 248 subjects with dyslipidemia were enrolled. Compared to the other 3 groups, subjects with severe OSA were older, male predominant and had higher smoking rate. In addition, subjects with severe OSA had higher body mass index, waist-hip ratio, blood pressure, and higher rates of overweight and obesity. Serum levels of fasting plasma glucose, glycated hemoglobin, LDL-C and CRP were all significantly higher. ASCVD prevalence was considerably higher in subjects with severe OSA. AHI event in the severe OSA group was up to 35.4 ± 5.1 events per hour which was significantly higher than the other groups (P < 0.05 for trend). Pearson correlation analysis showed that only LDL-C was positively correlated with AHI events (r = 0.685, P < 0.05). Logistic regression analysis revealed that in unadjusted model, compared to dyslipidemia plus no-OSA group (reference group), OSA enhanced ASCVD risk in subjects with dyslipidemia, regardless of OSA severity. After extensively adjusted for confounding variables, the odds of dyslipidemia plus mild-OSA was reduced to insignificance. While the effects of moderate- and severe-OSA on promoting ASCVD risk in subjects with dyslipidemia remained significant, with severe-OSA most prominent (odds ratio: 1.52, 95% confidence interval: 1.13-2.02). CONCLUSION: OSA combined dyslipidemia conferred additive adverse effects on cardiovascular system, with severe-OSA most prominent.

Mental distress among Liberian medical staff working at the China Ebola Treatment Unit: a cross sectional study.

BACKGROUND: Ebola virus outbreak in West Africa not only triggered a grave public health crisis, but also exerted and induced huge mental distress on medical staff, which would negatively influence epidemic control and social rebuilt furthermore. We chose the local medical staff working at the China Ebola Treatment Unit (ETU) to explore the severity of potential mental distress and involved potential causes. METHODS: A descriptive study using the Symptom Check List 90 - Revised (SCL90-R) questionnaire to assess psychological health status was conducted among 52 Liberian medical staff. Global indices, including Global Severity Index (GSI), Positive Symptom Total (PST) and Positive Symptom Distress Index (PSDI), and nine subscales based on 90 inquiry items were compared among gender, work duty and other subgroups. Data were analyzed using Graphpad Prism and SPSS software. RESULTS: Mental distress among participants was not very serious; only PSDI, paranoid ideation and interpersonal sensitivity numerically increased relative to changes in other categories. While male medics and those responsible for cleaning and disinfection showed significant increases in scores for psychological dimensions, such as obsessive-compulsive, anxiety, phobic anxiety, interpersonal sensitivity, paranoid ideation and positive symptom total. CONCLUSIONS: Data of this study implies that the psychological health status of medical staff within the special social environment of an Ebola treatment unit should warrant more attention.

Immunization with advanced glycation end products modified low density lipoprotein inhibits atherosclerosis progression in diabetic apoE and LDLR null mice.

BACKGROUND: Diabetes accelerates atherosclerosis through undefined molecular mechanisms. Hyperglycemia induces formation of advanced glycation end product (AGE)-modified low-density lipoprotein (LDL). Anti-AGE-LDL autoantibodies favor atherosclerosis (AS) progression in humans, while anti oxidized LDL immunization inhibits AS in hypercholesterolemic, non-diabetic mice. We here investigated if AGE-LDL immunization protects against AS in diabetic mice. METHODS: After diabetes induction with streptozotocin and high fat diet, both low density lipoprotein receptor (LDLR)-/- and apoE female mice were randomized to: AGE-LDL immunization with aluminum hydroxide (Alum) adjuvant; Alum alone; or PBS. RESULTS: AGE-LDL immunization: significantly reduced AS; induced specific plasma IgM and IgG antibodies; upregulated splenic Th2, Treg and IL-10 levels, without altering Th1 or Th17 cells; and increased serum high density lipoprotein(HDL) while numerically lowering HbA1c levels. CONCLUSIONS: Subcutaneous immunization with AGE-LDL significantly inhibits atherosclerosis progression in hyperlipidemic diabetic mice possibly through activation of specific humoral and cell mediated immune responses and metabolic control improvement.

[Effect of advanced glycation end products on the function and angiogenesis of adipose tissue-derived stem cells and the protective effect of danhong injection: an experimental study].

UNLABELLED: OBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test. RESULTS: Compared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05). CONCLUSION: clusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Depression and anxiety symptom network structure among patients with coronary heart disease and association with quality of life: protocol for a multicentre cross-sectional and prospective longitudinal study.

BACKGROUND: Anxiety and depression are critical mental health problems among persons with coronary heart disease (CHD). The range of symptoms is an important stressor for adverse cardiovascular events, and these symptoms can be involved in various ways during the course of CHD. However, the characteristics and mechanisms of comorbidity between the two mental states from the viewpoint of symptom interactions in patients with CHD remain unclear. Therefore, we aim to apply a symptom-oriented approach to identify core and bridge symptoms between anxiety and depression in a population with CHD and to identify differences in network structure over time and symptomatic link profiles. METHODS AND ANALYSIS: We designed a multicentre, cross-sectional, longitudinal study of anxiety and depression symptoms among patients with CHD. We will evaluate degrees of symptoms using the Generalized Anxiety Disorder Scale, the Patient Health Questionnaire and the WHO Quality of Life-Brief version. Patients will be followed up for 1, 3 and 6 months after baseline measurements. We will analyse and interpret network structures using R software and its packages. The primary outcomes of interest will include centrality, bridge connections, estimates, differences in network structures and profiles of changes over time. The secondary outcome measures will be the stability and accuracy of the network. By combining cross-sectional and longitudinal analyses, this study should elucidate the central and potential causative pathways among anxiety and depression symptom networks as well as their temporal stability in patients with CHD. ETHICS AND DISSEMINATION: The project conforms to the ethical principles enshrined in the Declaration of Helsinki (2013 amendment) and all local ethical guidelines. The ethics committee at the University of South China approved the study (Approval ID: 2023-USC-HL-414). The findings will be published and presented at conferences for widespread dissemination. TRIAL REGISTRATION NUMBER: ChiCTR2300075813.

An exploration of proactive health oriented symptom patterns in patients undergoing percutaneous coronary intervention with stent implantation: A mixed-methods study protocol.

BACKGROUND: Coronary Heart Disease (CHD) is one of the most prevalent chronic diseases worldwide. Currently, percutaneous coronary intervention (PCI) with stent implantation is the main clinical treatment for CHD, and patients can achieve better outcomes after stenting. However, adverse cardiovascular events continue to recur, ultimately failing to yield good results. Several symptoms exist after stenting and are associated with health outcomes. Little is known about the symptom patterns of patients during the different postoperative periods. Therefore, this study aims to explore the dynamics of symptoms and clarify the experiences of post-stenting in patients during different periods, which may help the delivery of more specific patient management and improve survival outcomes in the future. METHODS: A mixed method (quantitative/qualitative) design will be adopted. Longitudinal research, including surveys regarding three different periods, will be sued to describe the symptom patterns of patients undergoing PCI with stent implantation, clarifying their focused symptom problems during different time periods or in populations with different features. Qualitative individual interviews aim to understand the feelings, experiences, opinions, and health conditions of patients post-stenting, which can explain and supplement quantitative data. Quantitative data will be analyzed using descriptive statistics, latent class analysis (LCA), and latent translation analysis (LTA). Qualitative data will be analyzed using content analysis. DISCUSSION: This study is the first study to explore the symptom patterns and experiences of patients in various domains after stent implantation using a novel design including quantitative and qualitative methods, which will help the delivery of more specific patient management, reduce the recurrence of adverse cardiovascular events, and improve survival outcomes in the future. It is also meaningful to use PROMIS profile-57 to help patients to proactively focus on their health problems, promote health literacy, and incorporate active patient participation into health management, which is a successful transition from passive medical treatment to active management.