The circadian clock is crucial for the progression of cardiovascular diseases. Our previous studies showed that acrolein, an environmental pollutant, exacerbated atherosclerosis by reducing CLOCK/BMAL1 levels and disrupting circadian rhythm; in contrast, intermittent fasting (IF), a dietary regimen, ameliorated acrolein-induced atherosclerosis. In the current study, mice were administered acrolein at a dose of 3 mg/(kg·day) via drinking water and subjected to IF for 18 h (from 0:00 to 18:00). We observed that IF reduced the formation of aortic lesions accelerated by acrolein in ApoE^−/− mice. Upon exposure to acrolein, the expression of RelA, Il1b, and Tnf increased in the liver and heart tissues, but these changes were reversed by IF treatment. Notably, IF treatment upregulated the expression of adenosine monophosphate (AMP)-activated protein kinase catalytic subunit alpha-1 (AMPKα1), p-AMPKα1, and sirtuin 1 (SIRT1), while inhibiting acrolein-induced mitogen-activated protein kinase (MAPK) activation. Additionally, the expression of circadian genes Clock/Bmal1 was suppressed and disrupted by acrolein, whereas IF restored their expression. Moreover, consistent with the in vivo findings, short-term starvation in vitro, as a fasting cell model, alleviated the dysregulation of CLOCK/BMAL1 and upregulated SIRT1 expression by modulating the AMPK and reactive oxygen species (ROS)-MAPK pathways activated by acrolein. In summary, we demonstrated that IF suppressed the ROS-MAPK pathway but activated the AMPK pathway to enhance the expression of circadian clock genes, thereby ameliorating acrolein-induced atherogenesis, which may shed light on strategies for preventing cardiovascular diseases.

It is often challenging to diagnose acute myocardial infarction (AMI) in patients with elevated high-sensitivity cardiac troponin T (hs-cTnT) before observing a significant rise and/or fall in hs-cTnT. The current study aimed to identify an optimal cut-off to rule in AMI. A total of 76411 patients with elevated hs-cTnT were included. The predictive cut-off values for diagnosing ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation myocardial infarction (NSTEMI) were assessed using the area under the receiver operating characteristic curve (AUC). Among the patients, 50466 (66.0%) had non-cardiac diseases, 25945 (34.0%) had cardiac diseases, and 15502 (20.3%) had AMI, including 816 (1.1%) with STEMI and 14686 (19.2%) with NSTEMI. The median hs-cTnT level was 3788.0 ng/L in STEMI patients and 67.2 ng/L in NSTEMI patients. The optimal cut-off for diagnosing STEMI was 251.9 ng/L, with a sensitivity of 90.7%, specificity of 86.5%, and an AUC of 0.942; the optimal cut-off for diagnosing NSTEMI was 130.5 ng/L, with a sensitivity of 40.9%, specificity of 83.8%, and an AUC of 0.638. Collectively, optimizing the cut-off values for diagnosing STEMI and NSTEMI to 251.9 ng/L and 130.5 ng/L, respectively, demonstrated high accuracy in a large cohort of Chinese patients with elevated hs-cTnT.

A mutation in oxysterol-binding protein-like 2 (OSBPL2) has been identified as the genetic cause of autosomal dominant nonsyndromic hearing loss (DFNA67, Online Mendelian Inheritance in Man No. 616340). However, the pathogenesis of the OSBPL2 mutation in DFNA remains unclear. Our previous work showed that Osbpl2 deficiency impaired cell adhesion in auditory HEI-OC1 cells. In addition, loss of hair cells (HCs) and morphological abnormalities of HC stereocilia were detected in OSBPL2-knockout pigs, suggesting that OSBPL2 plays an important role in regulating the actin cytoskeleton in auditory cells. In the present study, we found that Osbpl2 deficiency inhibited the Rho/ROCK2 signaling pathway and downregulated phosphorylated ezrin-radixin-moesin (p-ERM), resulting in abnormal F-actin morphology in HEI-OC1 cells and stereociliary defects in mouse HCs. The present study demonstrates the underlying mechanism of OSBPL2 in regulating the actin cytoskeleton in HCs, contributing to a deeper understanding of the pathogenesis of OSBPL2 mutations in DFNA.

Primordial germ cells (PGCs), the precursors of oocytes and spermatozoa, are highly pluripotent. In recent years, the in vitro induction of human primordial germ cell-like cells (hPGCLCs) has advanced significantly. However, the stability and efficacy of obtaining hPGCLCs in vitro still require improvement. In the current study, we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) as the basal medium, supplemented with B27 and N2 (referred to as N2B27) in combination with four cytokines: bone morphogenetic protein 4, stem cell factor, epidermal growth factor, and leukemia inhibitory factor. The hPGCLCs induced under these conditions closely resembled PGCs from 4- to 5-week-old embryos at the transcriptomic level. Compared with traditional GK15 (GMEM supplemented with 15% Knockout™ Serum Replacement)-based induction conditions, the N2B27 system significantly increased the speed and efficacy of hPGCLC induction. RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis. The N2B27 medium promoted enhanced mitochondrial activity, enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species. Moreover, through gradient concentration experiments, we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions. In summary, we have established an optimized induction system that enhances the efficiency of hPGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.

The study examined the association between weekly mean temperature and influenza cases across 122 countries/regions (2014-2019) using a distributed lag non-linear model. We analyzed 3145206 cases of overall influenza (Flu-All), with influenza A (Flu-A) and influenza B (Flu-B) accounting for 73.49% and 26.51%, respectively. Within a lag of two weeks, Flu-All incidence demonstrated a bimodal temperature relationship, with peak relative risks (RR) of 6.02 (95% confidence interval [CI]: 1.92-20.77) at -8 ℃ and 3.08 (95% CI: 1.27-7.49) at 22 ℃. Flu-A exhibited a similar bimodal pattern, with RRs of 3.76 (95% CI: 2.39-5.91) at −8 ℃ and 2.08 (95% CI: 1.55-2.80) at 22 ℃. Flu-B demonstrated a single risk peak at 1 ℃ (RR = 4.48, 95% CI: 1.74-11.55). Subgroup analyses of climate zones revealed variations: tropical zones peaked at 12 ℃ (RR = 1.37, 95% CI: 1.08-1.74), while dry and temperate zones exhibited the highest risks at −5 ℃, with RRs of 4.49 (95% CI: 2.46-7.15) and 5.23 (95% CI: 3.17-8.64), respectively. Cold zones peaked at 1 ℃ (RR = 5.96, 95% CI: 3.76-9.43). Subgroup analyses of influenza transmission zones revealed variations: Africa showed a higher risk between 6 ℃ and 14 ℃, Asia showed a higher risk below 3 ℃, and Europe exhibited distinct risks of influenza peaks at −1 ℃ (Eastern Europe), 1 ℃ (Southwestern Europe), and −20 ℃ (Northern Europe). Elevated risks above 11 ℃ were identified in the Americas and Oceania. These findings establish a predictive framework for influenza outbreak preparedness by integrating regional temperature patterns with global climate variability.

The Optimal Management of Antithrombotic and Thrombolytic Agents-5 (OPTIMA-5) study demonstrated that a single bolus of half the standard dose of recombinant staphylokinase (r-SAK) before primary percutaneous coronary intervention (PCI) significantly improved the patency of the infarct-related artery in patients with ST-segment elevation myocardial infarction (STEMI), who were expected to undergo PCI within 120 min. The present study aimed to investigate the one-year clinical outcomes and the effect of the anti-r-SAK antibodies on a second r-SAK thrombolysis in OPTIMA-5 patients. The clinical outcome measured was major adverse cardiovascular events (MACE) within 360 days. Patients' anti-r-SAK antibody levels were determined on day 90 (± 7 days), day 180 (± 7 days), and day 360 (± 14 days) after thrombolysis, and in vitro r-SAK antibody neutralization experiments were performed to explore an optimal interval for a second r-SAK thrombolysis. Results showed that the MACE incidence was numerically lower in the r-SAK group compared with the normal saline (NS) group (14.0% vs. 20.0%, hazard ratio [HR] = 0.67, 95% confidence interval [CI]: 0.34-1.32; log-rank P = 0.245). The anti-r-SAK antibody levels in the r-SAK group decreased with time, but remained significantly higher than those in the NS group on day 90 (± 7 days) (2.96 ± 0.68 vs. 0.22 ± 0.53, P < 0.001), day 180 (± 7 days) (2.19 ± 0.74 vs. 0.44 ± 0.65, P < 0.001), and day 360 (± 14 days) (1.73 ± 0.97 vs. 0.37 ± 0.71, P < 0.001). The in vitro anti-r-SAK antibody neutralization experiments demonstrated that the thrombolysis rate decreased exponentially as the antibody titer increased from 1.90 to 2.20 (67.80% ± 14.19% vs. 44.32% ± 21.54%, P < 0.0001). Therefore, for STEMI patients who are expected to undergo PCI within 120 min, a single bolus of half-dose r-SAK before primary PCI may reduce the one-year MACE risk. The anti-r-SAK antibody persists over one year, and a second r-SAK thrombolysis may not be indicated until at least one year after the first administration, if necessary.

Silicosis is an occupational lung disease caused by prolonged exposure to silica dust in the workplace. It has a complex pathogenesis and currently lacks effective treatments. Homoharringtonine (HHT) is a natural compound approved for the treatment of acute myeloid leukemia, but its effects on silicosis remain unclear. In the present study, we constructed a mouse model of silica (SiO₂)-induced pulmonary fibrosis and evaluated the preventive and therapeutic effects of HHT. The results showed that HHT significantly attenuated the progression of SiO₂-induced pulmonary fibrosis in mice. We then used MRC-5, a human lung fibroblast cell line, to explore the mechanisms underlying HHT's inhibitory effects in vitro and found that HHT significantly inhibited the activation and migratory capacity of MRC-5 cells. Mechanistically, these effects were mediated by enhanced ubiquitination and degradation of the CCR1 protein. Furthermore, HHT exhibited favorable biocompatibility in vivo, and its preventive and therapeutic effects were validated in SiO₂-treated mice. Collectively, the current study demonstrates that HHT shows significant potential as a therapeutic agent for silicosis by targeting CCR1 and the PI3K/AKT/mTOR signaling pathway, highlighting it as a promising candidate for clinical translation for silicosis treatment.

This study aimed to evaluate the association of a healthy lifestyle pattern with mortality risk among patients with type 2 diabetes mellitus (T2DM). Data were derived from a prospective cohort study enrolling 13776 Chinese patients with T2DM. A healthy lifestyle pattern was constructed based on six lifestyle factors, including smoking status, alcohol consumption, dietary habits, physical activity, sedentary time, and sleep duration. Multivariate Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for all-cause and cause-specific mortality. During a median follow-up of 9.78 years, 2497 deaths were recorded. Compared with T2DM patients with a lifestyle pattern scoring 0-2, those scoring 5-6 had a 40% lower risk for all-cause mortality (HR = 0.60, 95% CI: 0.52-0.69), a 33% lower risk for cardiovascular disease mortality (HR = 0.67, 95% CI: 0.52-0.86), and a 25% lower risk for cancer mortality (HR = 0.75, 95% CI: 0.58-0.97). Additionally, we found that the association between the lifestyle pattern and all-cause mortality risk was stronger in females than in males (P for interaction < 0.05). In conclusion, adherence to a healthy lifestyle pattern is associated with a decreased risk of all-cause, cardiovascular disease, and cancer mortality. These findings have important implications for reducing premature mortality among patients with T2DM.