The MFF-SIRT1/3 axis, regulated by miR-340-5p, restores mitochondrial homeostasis of hypoxia-induced pulmonary artery smooth muscle cells.

Mitochondrial dynamics and quality control play a central role in the maintenance of the proliferation-apoptosis balance, which is closely related to the progression of pulmonary arterial hypertension (PAH). However, the exact mechanism of this balance remains unknown. Pulmonary artery smooth muscle cells (PASMCs) were cultured in hypoxia condition for constructing a PAH model in vitro. The expression of genes and proteins were determined by qRT-PCR and western bolt assays. Cell proliferation-apoptosis balance were tested by MTT, EdU and TUNEL assays. The mitochondrial functions were assessed by flow cytometry, JC-1, Mito tracker red staining, and corresponding kits. Besides, the molecular interaction was validated by dual-luciferase reporter assay. MFF was overexpressed in hypoxia-treated PAMSCs. Knockdown of MFF significantly repressed the excessive proliferation but enhanced cell apoptosis in hypoxia-treated PAMSCs. Moreover, MFF silencing improved mitochondrial function of hypoxia-treated PAMSCs by increasing ATP production and decreasing ROS release and mitochondrial fission. Mechanistically, MFF was a directly target of miR-340-5p, and could negatively regulate SIRT1/3 expression. Subsequently, functional rescue assays showed that the biological effects of MFF in hypoxia-treated PAMSCs were negatively regulated by miR-340-5p and depended on the regulation on SIRT1/3 pathway. These results provided evidences that miR-340-5p regulated MFF-SIRT1/3 axis to improve mitochondrial homeostasis and proliferation-apoptosis imbalance of hypoxia-treated PAMSCs, which provided a theoretical basis for the prevention and treatment of PAH.

Renal denervation ameliorates cardiac metabolic remodeling in diabetic cardiomyopathy rats by suppressing renal SGLT2 expression.

This study aimed to investigate the effects of renal denervation (RDN) on diabetic cardiomyopathy (DCM) and explore the related mechanisms. Male Sprague-Dawley rats were fed high-fat chow and injected with low-dose streptozotocin to establish a DCM model. Six rats served as controls. The surviving rats were divided into three groups: control group, DCM group and DCM + RDN group. RDN surgery was performed in the fifth week. At the end of the experiment, all rats were subjected to 18F-FDG PET/CT and metabolic cage studies. Cardiac function and structure were evaluated by echocardiography and histology. Myocardial substrate metabolism and mitochondrial function were assessed by multiple methods. In the 13th week, the DCM rats exhibited cardiac hypertrophy and interstitial fibrosis accompanied by diastolic dysfunction. RDN ameliorated DCM-induced cardiac dysfunction (E/A ratio: RDN 1.07 ± 0.18 vs. DCM 0.93 ± 0.12, P < 0.05; E/E' ratio: RDN 10.74 ± 2.48 vs. DCM 13.25 ± 1.99, P < 0.05) and pathological remodeling (collagen volume fraction: RDN 5.05 ± 2.05% vs. DCM 10.62 ± 2.68%, P < 0.05). Abnormal myocardial metabolism in DCM rats was characterized by suppressed glucose metabolism and elevated lipid metabolism. RDN increased myocardial glucose uptake and oxidation while reducing the absorption and utilization of fatty acids. Meanwhile, DCM decreased mitochondrial ATP content, depolarized the membrane potential and inhibited the activity of respiratory chain complexes, but RDN attenuated this mitochondrial damage (ATP: RDN 30.98 ± 7.33 µmol/gprot vs. DCM 22.89 ± 5.90 µmol/gprot, P < 0.05; complexes I, III and IV activity: RDN vs. DCM, P < 0.05). Furthermore, both SGLT2 inhibitor and the combination treatment produced similar effects as RDN alone. Thus, RDN prevented DCM-induced cardiac dysfunction and pathological remodeling, which is related to the improvement of metabolic disorders and mitochondrial dysfunction.

Neuronal nitric oxide synthase in dorsal raphe nucleus mediates PTSD-like behaviors induced by single-prolonged stress through inhibiting serotonergic neurons activity.

The pathogenesis of post-traumatic stress disorder (PTSD) remains largely unclear. A large body of evidence suggests that the abnormal level of serotonin (5-HT) is closely related to the onset of PTSD. Several reports reveal that nitric oxide (NO) affects extracellular 5-HT levels in various brain regions, but no consistent direction of change was found and the underlying mechanisms remain unknown. The most of serotonergic neurons in dorsal raphe nucleus (DRN), a major source of serotonergic input to the forebrain, co-expresses neuronal nitric oxide synthase (nNOS), a synthase derived nitric oxide (NO) in the central nervous system. Here, we found that the excessive expression of nNOS and thereby the high concentration of NO followed by single-prolonged stress (SPS) caused suppression of the activity of DRN 5-HT neurons, inducing PTSD-like phenotype including increased anxiety-like behaviors, enhanced contextual fear memory, and fear generalization. Our study uncovered an important role of DRN nNOS-NO pathway in the pathology of PTSD, which may contribute to new understanding of the molecular mechanism of PTSD.

Effects of Angiotensin Receptor Neprilysin Inhibitors on Inducibility of Ventricular Arrhythmias in Rats with Ischemic Cardiomyopathy.

The aims of the present study were to investigate the effects of angiotensin receptor neprilysin inhibitors (ARNi) on the susceptibility of ventricular arrhythmias (VAs) in rats with myocardial infarction (MI) and to explore the related mechanisms.A total of 32 adult male Sprague-Dawley rats were divided into 3 groups: a control group, MI group, and MI+ARNi group. MI was generated by ligation of the left anterior descending coronary artery. ARNi was given at 68 mg/kg/day for 4 weeks after MI surgery. At 4 weeks after MI, electrical programmed stimulation (EPS) was performed in all groups for the evaluation of VAs, and echocardiography was used to evaluate cardiac function. Indicators of sympathetic neural remodeling and cardiac remodeling were detected to further explore the related mechanisms.Four weeks after MI, rats in the ARNi group exhibited low susceptibility of VAs in comparison with that in the MI group, which was coincident with the attenuation of sympathetic nerve remodeling, amelioration of cardiac fibrosis, and regulation of Cx43 expression.ARNi is effective in reducing VAs in rats with ischemic cardiomyopathy, which is associated with attenuating sympathetic nerve remodeling and myocardial fibrosis.

[Prevalence of human papilloma virus infection and its risk factors in some rural areas of Jiangsu, China].

OBJECTIVE: To investigate human papilloma virus (HPV) infection in the male genital tract and its risk factors in some rural areas of Jiangsu Province. METHODS: This study included 398 men from six rural areas in Jiangsu Province, whose female partners, based on the results of cervical cytological examination, were divided into a normal (n = 104), a cervical intraepithelial neoplasia grade Ⅰ (CIN-Ⅰ, n = 100), a CIN-Ⅱ (n = 95), and a CIN-Ⅲ group (n = 99). We examined the male subjects for genital warts and other lesions, collected urethral swab samples for HPV detection, and obtained their sociodemographic data by questionnaire investigation. RESULTS: No megascopic lesions were observed in the genitals of the 398 participants. The total prevalence rate of HPV infection was 11.31% and that of high-risk HPV was 8.54%. Logistic regression analysis showed that daily cleaning of the genitals significantly decreased the risk of HPV infection (OR = 3.030, P = 0.003). CONCLUSIONS: There is a relatively high prevalence rate of recessive infection of genital HPV among the seemingly healthy males in the rural area of Jiangsu Province. Daily cleaning of the genitals may be a protective measure against HPV infection.

A Meta-analysis of Central Corneal Thickness Changes With Overnight Orthokeratology.

OBJECTIVES: To assess the change of central corneal thickness (CCT) in the treatment of orthokeratology in patients with myopia. METHODS: A systematic search of all relevant studies published through April 2014 was conducted, and 95% confidence intervals (CI) of CCT change were calculated. Random or fixed-effects models were used according to heterogeneity. Publication bias of the articles was evaluated using funnel plots and Begg test. RESULTS: A total of 10 studies with 239 patients (339 eyes) from clinical studies were included. Central corneal thickness reduced significantly from 1 day to 1 week by 5.73 µm (95% CI, 1.75-9.70 µm; P=0.005), and a significant mean reduction of 5.89 µm also occurred from 1 day to 1 month (95% CI, 3.50-8.29 µm; P<0.001). No significant reduction was found between 1 week and 1 month (P=0.32). CONCLUSIONS: Our meta-analysis demonstrated that most reduction of CCT occurred during the first week and remained thinner for 1 month. Further randomized controlled trials with larger sample sizes, standardized outcome measurements, and different follow-up periods are warranted to find the precise change.

Spliced XBP1 promotes macrophage survival and autophagy by interacting with Beclin-1.

Macrophage autophagy plays an important role in the development of atherosclerosis, but the precise mechanism mediating this process is unclear. The potential role of the X-box binding protein 1 (XBP1), a crucial transduction factor that is involved in endoplasmic reticulum stress and the unfolded protein response, in bone marrow-derived macrophage autophagy is unknown. This study mainly explores the roles of XBP1 mRNA splicing in bone marrow-derived macrophage autophagy. The present study shows that the transient overexpression of spliced XBP1 via adenovirus-mediated gene transfer induces autophagy and promotes proliferation in bone marrow-derived macrophages via the down-regulation of Beclin-1, but that the sustained overexpression of spliced XBP1 leads to apoptosis. When XBP1 is down-regulated in bone marrow-derived macrophages using siRNA, rapamycin-induced autophagosome formation is ablated. Furthermore, we have detected the overexpression of XBP1 in areas of atherosclerotic plaques in the arteries of ApoE-/- mice. These results demonstrate that XBP1 mRNA splicing plays an important role in maintaining the function of bone marrow-derived macrophages and provide new insight into the study and treatment of atherosclerosis.

[Protective effect of thymosin ß4 against oxidative damage in cultured rabbit corneal epithelial cells].

OBJECTIVE: The aim of this study was to investigate the effects of thymosin ß4 (Tß4) against oxidative damage in rabbit corneal epithelial cells in vitro. METHODS: Experimental study. Primary cultures of rabbit corneal epithelial cells were isolated and cultured from cornea tissue explants. Cell morphology was observed by phase-contrast microscope. The expression of keratin 12 and connexin 43 in cultured cells were examined with RT-PCR. The cultures were divided into 4 groups: control group, Tß4 group, hydrogen peroxide (H2O2) group, and H2O2+Tß4 group. The cell viability of cultured corneal epithelial cells was examined by MTT assay. TUNEL staining was used to detect the apoptotic cells. ROS levels were estimated by DCFH-DA using fluorescent microscopy. Cell migration was measured using the scratch wound technique. Data were analyzed using one-way analysis of variance (ANOVA) and secondary analysis for significance with post Hoc tests. RESULTS: The morphology of cultured cells was round, ovoid, polygonal or paving stone appearance. The results of RT-PCR showed that cultured corneal epithelial cells expressed both keratin 12 and connexin 43, the characteristic genes of corneal epithelial cells. The cell viability in H2O2 group was significantly reduced than that in control group (67.20% ± 5.87% vs. 100.00% ± 9.99%, P = 0.000); the cell viability was significantly improved in Tß4+ H2O2 group than that in H2O2 group (83.42% ± 7.23% vs. 67.20% ± 5.87%, P = 0.023) . Cell migration was significantly increased in Tß4 group compared with the controls (117.6% ± 2.22% vs. 100.00% ± 4.06%, P = 0.005) ; Compared with H2O2 group, cell migration was significant increase in H2O2 + Tß4 group (96.57% ± 8.22% vs. 64.38% ± 11.08%, P = 0.000) . Compared with the control group, the intracellular ROS level of the H2O2 group was significantly increased (234.42% ± 22.15% vs. 100.00% ± 5.28%, P = 0.000) . Intracellular ROS level of H2O2+Tß4 group was significantly decreased as compared with the H2O2 group (163.26% ± 10.53% vs. 234.42% ± 22.15%, P = 0.000). TUNEL staining indicated that Tß4 treatment markedly inhibited H2O2-induced apoptosis in cultured corneal epithelial cells. CONCLUSIONS: Tß4 has a strong protection effect against oxidative damage induced by H2O2 in corneal epithelial cells through promoting cell growth and cell migration, and the underlying mechanisms may due to the antioxidation and anti-apoptotic effects of Tß4.

Prognostic value of ultra-sensitive pregnancy associated plasma protein-A in patients with acute coronary syndrome.

OBJECTIVE: To investigate the prognostic value of ultra-sensitive pregnancy associated plasma protein-A (PAPP-A) level in the early phase of acute coronary syndrome (ACS) attack. METHODS: Patients diagnosed as ACS were enrolled and the level of circulatory PAPP-A was measured within 12 hours after ACS attack. The patients were followed at the time of 1st, 6th, and 12th months post-ACS attack in order to observe the incidence of the cardiovascular adverse events. According to the highest quintile, the patients were divided into 2 groups: high level (≥26.08 µg/L) group and low level (<26.08 µg/L) group, to evaluate the association between the level of PAPP-A and the incidence of the cardiovascular events. RESULTS: Compared with the low level group, the incidence of the composite outcome is significantly increased in the high level group, and the values of OR are 4.76, 4.38, 3.75 for 1st, 6th, 12th months respectively (P=0.000). For myocardial infarction (MI) + cardiac death (CD) the values of OR were 9.81, 6.08, 4.12 (P<0.01). Multivariate logistic regression analysis demonstrates that PAPP-A was an independent risk factor for the cardiovascular adverse events in the early, median, and late phase of ACS (P<0.05). CONCLUSION: In the early phase of ACS attack, the elevation of PAPP-A is an independent risk factor for the occurrence of cardiovascular adverse events.

Hsa_circ_0016070/micro-340-5p Axis Accelerates Pulmonary Arterial Hypertension Progression by Upregulating TWIST1 Transcription Via TCF4/ß-Catenin Complex.

Background Hypoxia is considered a major leading cause of pulmonary hypertension (PH). In this study, the roles and molecular mechanism of circ_0016070 in PH were studied. Methods and Results The expression of circ_0016070 in serum samples, human pulmonary artery smooth muscle cells and hypoxia/monocrotaline-treated rats was determined by real-time quantitative polymerase chain reaction. Cell viability, migration, and apoptosis were analyzed by Cell Counting Kit-8, wound healing, flow cytometry, and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assays, respectively. The molecular interactions were validated using RNA immunoprecipitation, chromatin immunoprecipitation, and dual luciferase reporter assays. The levels of phenotype switch-related proteins were evaluated by Western blot and immunohistochemistry. The pathological characteristics were assessed using hematoxylin and eosin staining. circ_0016070 was highly expressed in the serum samples, hypoxia-induced pulmonary artery smooth muscle cells and pulmonary arterial tissues of PH rats. Downregulation of circ_0016070 ameliorated the excessive proliferation, migration, vascular remodeling, and phenotypic transformation but enhanced cell apoptosis in the PH rat model. In addition, micro (miR)-340-5p was verified as a direct target of circ_0016070 and negatively regulated TCF4 (transcription factor 4) expression. TCF4 formed a transcriptional complex with ß-catenin to activate TWIST1 (Twist family bHLH transcription factor 1) expression. Functional rescue experiments showed that neither miR-340-5p inhibition nor TWIST1 or TCF4 upregulation significantly impeded the biological roles of circ_0010670 silencing in PH. Conclusions These results uncovered a novel mechanism by which circ_0016070 play as a competing endogenouse RNA of miR-340-5p to aggravate PH progression by promoting TCF4/ß-catenin/TWIST1 complex, which may provide potential therapeutic targets for PH.