OBJECTIVES: To investigate the protective effects of dexmedetomidine (DEX) against heat stress (HS)-induced oncosis in human skeletal muscle cells (HSKMCs) and its underlying mechanisms. METHODS: A HSKMC model of HS-induced oncosis were established by 43 ℃ water bath for 4 h, and the effects of treatments with 30 μmol/L DEX, ML385 (a Nrf2 inhibitor) +DEX, si-Nrf2+HS, and si-Nrf2+DEX prior to modeling on cell viability was assessed using CCK-8 assay. Oncosis characteristics were evaluated using transmission electron microscopy and Annexin V-FITC/PI flow cytometry. The oxidative stress markers (GSH, GSH-Px, MDA, SOD and ROS), mitochondrial membrane potential, energy metabolism, and inflammatory cytokines (TNF-α, IL-6 and IL-1β) in the cells were quantified using standard kits, and the expressions of porimin, caspase-3 and Nrf2 pathway proteins were analyzed using Western blotting and qRT-PCR. RESULTS: HS induced typical oncotic features in HSKMCs including organelle swelling and cytoplasmic vacuolization. DEX pretreatment significantly attenuated these changes, reduced Annexin V⁺/PI⁺ cell ratio and cellular porimin expression, and lowered the levels of ROS and MDA while restoring GSH and SOD levels. DEX pretreatment also significantly increased the mitochondrial membrane potential and ATP level, upregulated the expressions of Nrf2, p-Nrf2, HO-1 and NQO1, and suppressed the expressions of TNF-α, IL-6 and IL-1β. The protective effects of DEX were obviously attenuated by interventions with ML385 or si-Nrf2. CONCLUSIONS DEX mitigates HS-induced HSKMC oncosis by activating the Nrf2/HO-1 pathway to relieve oxidative stress, mitochondrial dysfunction, and inflammatory responses.
Humans ; Dexmedetomidine/pharmacology* ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress/drug effects* ; Heat-Shock Response/drug effects* ; Signal Transduction/drug effects* ; Membrane Potential, Mitochondrial ; Muscle, Skeletal/cytology* ; Heme Oxygenase-1/metabolism* ; Apoptosis/drug effects*

Objective:To investigate the impact of receptor-interacting protein kinase 3 (RIPK3) on major adverse cardiovascular events (MACE) in patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI), as well as the predictive performance of RIPK3 combined with traditional cardiovascular risk factors.Methods:This study was a single-center prospective cohort study. It included patients with AMI who underwent PCI at Peking Union Medical College Hospital between September 2017 and November 2017. Baseline clinical data were collected, and plasma samples were obtained 6 hours after PCI to measure RIPK3 levels. Follow-up was conducted via outpatient visits or phone calls to record the occurrence of MACE, including cardiovascular death, hospitalization for heart failure, and vascular events (recurrent AMI or stroke). The predictive performance of RIPK3, traditional cardiovascular risk factors and their combination for MACE was compared using receiver operating characteristic (ROC) curves. Patients were divided into low- and high-RIPK3 level groups based on the optimal cutoff value of RIPK3. Multivariate Cox proportional hazards regression analysis was used to assess the impact of RIPK3 levels on MACE after PCI in AMI patients. Kaplan-Meier survival curves were plotted, and the log-rank test was used to compare MACE incidence between the low-and high-RIPK3 groups.Results:A total of 103 AMI patients who underwent PCI were included, aged 63.0 (56.0, 69.0) years, and 83 (80.6%) were male. The follow-up time was 5.17 (2.81, 5.17) years, during which 44 patients (42.7%) experienced MACE. The ROC curve analysis showed that the area under the curve ( AUC) for traditional cardiovascular risk factors was 0.68 (95% CI: 0.58-0.78), while the AUC for plasma RIPK3 was 0.72 (95% CI: 0.62-0.82). The combined AUC for traditional risk factors and RIPK3 was 0.75 (95% CI: 0.65-0.85). Multivariate Cox proportional hazards regression analysis indicated that plasma RIPK3 level is greater than or equal to the optimal cutoff value of 440.9 μg/L ( HR=3.31, 95% CI: 1.53-8.30, P=0.005) was an independent risk factor for MACE in AMI patients after PCI. Kaplan-Meier survival analysis demonstrated that the high-RIPK3 group had a significantly higher risk of MACE after PCI compared to the low-RIPK3 group (log-rank P=0.006). Conclusions:Elevated plasma RIPK3 level is an independent risk factor for MACE in AMI patients after PCI. Plasma RIPK3 combined with traditional cardiovascular risk factors can more effectively predict the occurrence of MACE in AMI patients after PCI. AMI patients with RIPK3≥440.9 μg/L have a higher risk of MACE after PCI.

Objective:To explore the relationship between retinol-binding protein (RBP) levels and disease severity in patients with transthyretin cardiac amyloidosis (ATTR-CA), as well as its impact on therapeutic response to tafamidis.Methods:This retrospective study utilized data from the China National Rare Disease Registry System and included ATTR-CA patients treated with tafamidis between January 2018 and September 2022. Patients were stratified into two groups based on baseline RBP levels: the normal RBP group (≥36 mg/L) and the reduced RBP group (<36 mg/L). Baseline characteristics and clinical data after one year of treatment were collected and compared between the groups. Within the reduced RBP group, patients were further subclassified by changes in RBP levels after treatment (ΔRBP=post-treatment RBP-baseline RBP) into ΔRBP>0 and ΔRBP<0 subgroups. Worsening of global longitudinal strain (GLS) after treatment was defined as the primary outcome, logistic regression analysis was used to identify risk factors influencing therapeutic response to tafamidis in ATTR-CA patients.Results:A total of 52 ATTR-CA patients were included (aged (58.5±12.0) years, 46 males (88%)). Among 39 patients who completed one-year tafamidis treatment, no statistically significant difference was observed in RBP levels post-treatment versus baseline ((27.0±14.3) mg/L vs. (25.9±15.4) mg/L, P=0.261). Compared to the normal RBP group, the reduced RBP group had significantly higher estimated glomerular filtration rate-adjusted N-terminal pro-B-type natriuretic peptide levels (2 316.0 (1 161.5, 6 027.8) ng/L vs. 806.2 (349.5, 1 735.8) ng/L), higher left ventricular mass index ((164.4±46.5) g/m2 vs. (123.9±31.8) g/m2), and lower left ventricular ejection fraction ((50.8±11.3)% vs. (58.8±6.2)%) (all P<0.05). Among 31 patients in the reduced RBP group who completed one-year tafamidis treatment, 23 were classified as ΔRBP>0 and 8 as ΔRBP<0. The ΔRBP<0 group exhibited greater GLS worsening than the ΔRBP>0 group (0.7 (-0.1, 1.4)% vs. -0.4 (-1.4, 0.2)%, P=0.027). Multivariate logistic regression analysis revealed that ΔRBP<0 was an independent risk factor for GLS worsening ( OR=8.584, 95%CI 1.186-62.150, P=0.033) in ATTR-CA patients. Conclusion:ATTR-CA patients with reduced RBP levels exhibit more severe left ventricular structural and functional impairment compared to those with normal RBP levels. Decline in RBP during treatment (ΔRBP<0) is associated with poorer response to tafamidis treatment. Monitoring RBP dynamics may assist clinicians in assessing disease severity and therapeutic response in ATTR-CA patients.

Objective:To preliminarily evaluate the lumen gain of sonodynamic therapy (SDT) mediated by sinoporphyrin sodium at carotid and femoral atherosclerotic plaque sites, and to assess whether concomitant statin use, lesion location, plaque echogenicity/type, and baseline stenosis severity modify the therapeutic response.Methods:This single-center, prospective, exploratory pilot clinical study enrolled patients with peripheral artery disease who attended the outpatient cardiology clinic of the First Affiliated Hospital of Harbin Medical University between February and September 2016. All enrolled patients received optimized oral medical therapy in combination with a single session of SDT. Vascular evaluation was performed using color Doppler ultrasound before treatment and 1 and 4 weeks after treatment. The primary efficacy endpoint was the percent change from baseline in luminal diameter stenosis at the site of the atherosclerotic plaque (%Δ) at week 4, while the secondary efficacy endpoint was %Δ at week 1. Subgroup analyses were conducted according to prior statin use, plaque location, plaque characteristics, and baseline degree of luminal stenosis.Results:A total of 24 patients, aged (70.7±2.2) years were enrolled. There were 20 (83%) males. Compared to baseline, luminal diameter stenosis at the plaque site reduced by week 4 ((50.1±1.2)% vs. (57.2±1.1)%, P<0.001), %Δ was(12.32±1.05)%; and luminal diameter stenosis also reduced by week 1 ((51.7±1.2)% vs. (57.2±1.1)%, P<0.001)), %Δ was(9.61±0.85)%. In subgroup analyses, the treatment effect on diameter stenosis was independent of prior statin use; SDT reduced stenosis in both carotid and femoral plaques; with superior efficacy observed in hypoechoic and mixed-echo plaques; and efficacy was observed across mild, moderate, and severe baseline stenosis categories (all P<0.05). Conclusion:In this single-center pilot study, SDT demonstrates therapeutic efficacy across mild, moderate, and severe stenoses, as well as in hypoechoic and mixed-echo plaques, showing potential to rapidly promote luminal gain at carotid and femoral atherosclerotic plaque sites.

Objective:To investigate the effect and underlying mechanism of sonodynamic therapy (SDT) on inflammation-related atrial fibrillation (AF) susceptibility.Methods:Lipopolysaccharide (LPS)-stimulated mouse and HL-1 mouse atrial myocyte models were used. (1) In vivo study: experimental groups included control, LPS, LPS+SDT, and SDT groups, with 20 mice in each group. Atrial fibrillation inducibility and duration were assessed by electrical stimulation. Western blot was used to analyze atrial expression of NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), interleukin (IL)-1β, and IL-18. Immunohistochemistry was used to detect calcium voltage-gated channel subunit alpha1 C (CACNA1C) expression. (2) In vitro study: cell counting kit-8 (CCK-8) and Western blot were used to determine the optimal and safe LPS concentration. The safe incubation condition for the sonosensitizer sinoporphyrin sodium was determined by CCK-8 and fluorometry. An LPS-induced inflammatory model in HL-1 atrial myocytes was used, with experimental groups including control, LPS, LPS+SDT, LPS+sinoporphyrin sodium, and LPS+ultrasound groups. NLRP3 was overexpressed using plasmid transfection, with experimental groups including control, NLRP3 plasmid, negative control plasmid, and NLRP3 plasmid+SDT groups. SDT was applied to LPS-stimulated or NLRP3-overexpressing HL-1 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to measure mRNA and protein levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Cleaved Caspase-1, IL-1β, IL-18, and CACNA1C. The NLRP3 inhibitor MCC950 was used to validate the relationship of NLRP3 and CACNA1C. The experimental groups included control, LPS, LPS+MCC950, and MCC950 groups. Intracellular reactive oxygen species (ROS) levels were detected using the probe DCFH-DA, and the ROS scavenger N-acetyl-L-cysteine (NAC) was used to test if the effects of SDT was ROS-dependent.Results:(1) In vivo: The LPS+SDT group exhibited a lower incidence of atrial fibrillation induction and a shorter duration of atrial fibrillation compared to the LPS group(both P<0.05). Protein expression levels of NLRP3 and IL-1β were lower than those in the LPS group (all P<0.05), while the expression of CACNA1C subunit tended to increase relative to the LPS group ( P>0.05). (2) In vitro: The safe concentration of LPS for administration was ≤20 μg/ml, with an optimal pro-inflammatory concentration of 4 μg/ml. The safe concentration of sinoporphyrin sodium for administration was 0.4 μmol/L, with an optimal incubation time of 4 hours. Compared to the LPS group or NLRP3 plasmid group, the LPS+SDT group or NLRP3 plasmid+SDT group exhibited lower expression levels of NLRP3, ASC, Cleaved Caspase-1, IL-1β, and IL-18, and higher mRNA and protein levels of CACNA1C (all P<0.05). The LPS+MCC950 group had higher CACNA1C protein expression than the LPS group ( P<0.05). SDT increased intracellular ROS levels, and NAC blocked the regulatory effects of SDT on NLRP3 and CACNA1C. Conclusion:SDT reduces atrial fibrillation susceptibility in mice by inhibiting NLRP3 inflammasome activation in atrial cardiomyocytes, thereby upregulating the L-type calcium channel subunit CACNA1C.

BACKGROUND:The induced membrane technique(Masquelet technique)is a novel two-stage surgical approach for the reconstruction of large bone defects,gaining increasing popularity in clinical applications.However,the precise mechanism underlying its bone defect repair is still not fully understood.OBJECTIVE:To review the background,repair mechanism and advantages of the induced membrane technique,the characteristics of the induced membrane,membrane-bone graft communication,selection of animal models,types and morphology of bone cement,the effects of loaded antibiotics on the induced membrane,choice of fixation methods,and bone tissue engineering materials to provide new insights for the future treatment of critical-sized bone defects and the improvement of the induced membrane technique.METHODS:A literature search was conducted in PubMed,Web of Science,and CNKI databases,covering publications from 1986 to 2024.A total of 890 references were retrieved.Manual screening and analysis were performed based on inclusion criteria related to the fundamental research of induced membrane technique,excluding those with poor relevance to the topic and duplicates.The included literature comprised original experimental studies,reviews,meta-analyses and other relevant publications.Finally,72 articles were included for summary and analysis.RESULTS AND CONCLUSION:(1)The mechanism underlying the bone defect repair using this technique remains unclear,but both the membrane and bone grafting are indispensable.(2)The induced membrane is a distinctively layered tissue rich in various bone-forming related cells,growth factors,and blood vessels,with its vascularization and secretion of growth factors dynamically changing overtime.(3)In terms of animal model selection,sheep are more similar to humans in anatomical structure,weight-bearing patterns,and bone remodeling.However,rats are more suitable considering their lower feeding costs,easier handling,and shorter modeling period.(4)Polymethyl methacrylate is not the only material that can be used to induce a biomembrane,and there may be more suitable materials capable of inducing higher-quality biomembranes.The recommended dose of antibiotics(primarily vancomycin)is 1-4 g per 40 g polymethyl methacrylate.(5)For animal fixation,especially in rats,the use of steel plates is more widespread,providing a more reliable and reproducible fixation method.(6)In the future,there is potential for new materials to replace autogenous bone and enhance the bone repair capabilities of the Masquelet technique.

In recent years, non-invasive brain stimulation has been widely used in clinical rehabilitation due to its safe and non-invasive features.Particularly for stroke patients, this technique has become a new type of rehabilitation by virtue of its role in improving gait disorders, aphasia, dysphagia, and other complications. Previous research on stroke rehabilitation mainly focused on the cerebral cortex, while the recent evidences suggest that the cerebellum, with its rich synaptic plasticity and extensive structural and functional connectivity with the brain, has emerged as a promising target for non-invasive intervention in stroke rehabilitation. Non-invasive stimulation of the cerebellum is expected to improve patients' impaired function and quality of life. This review discusses the possible mechanisms and clinical applications of non-invasive cerebellar stimulation technology in regulating stroke rehabilitation, aiming to provide new treatment directions and application value for individualized stroke rehabilitation.

The cerebellum plays a critical role not only in motor control but also in reward processing, emotional regulation, and addictive behaviors.Addictive behaviors alter the cerebellum’s neurotransmitter systems and synaptic connections, leading to dysfunction and reinforcing the persistence of addiction.This review summarizes the anatomical and functional foundations of the cerebellum in the reward circuitry and explores how cerebellar synaptic plasticity influences brain function in both substance and behavioral addictions. Furthermore, the potential role of the cerebellum in the progress of addiction and its clinical application prospects in addiction treatment are also discussed.

In recent years, non-invasive brain stimulation has been widely used in clinical rehabilitation due to its safe and non-invasive features.Particularly for stroke patients, this technique has become a new type of rehabilitation by virtue of its role in improving gait disorders, aphasia, dysphagia, and other complications. Previous research on stroke rehabilitation mainly focused on the cerebral cortex, while the recent evidences suggest that the cerebellum, with its rich synaptic plasticity and extensive structural and functional connectivity with the brain, has emerged as a promising target for non-invasive intervention in stroke rehabilitation. Non-invasive stimulation of the cerebellum is expected to improve patients' impaired function and quality of life. This review discusses the possible mechanisms and clinical applications of non-invasive cerebellar stimulation technology in regulating stroke rehabilitation, aiming to provide new treatment directions and application value for individualized stroke rehabilitation.

The cerebellum plays a critical role not only in motor control but also in reward processing, emotional regulation, and addictive behaviors.Addictive behaviors alter the cerebellum’s neurotransmitter systems and synaptic connections, leading to dysfunction and reinforcing the persistence of addiction.This review summarizes the anatomical and functional foundations of the cerebellum in the reward circuitry and explores how cerebellar synaptic plasticity influences brain function in both substance and behavioral addictions. Furthermore, the potential role of the cerebellum in the progress of addiction and its clinical application prospects in addiction treatment are also discussed.