Immune escape drives cancer progression and therapy resistance, yet its prognostic role and impact on the tumor immune microenvironment in thyroid cancer remain unclear. We integrated single-cell and bulk RNA sequencing data to systematically characterize immune escape and its clinical significance. scRNA-seq analysis characterized cellular heterogeneity and quantified immune escape activity via AUCell. A prognostic gene signature was constructed from differential expression analysis combined with univariate Cox and LASSO regression, and validated using Kaplan-Meier and time-dependent ROC analyses. The immune landscape was profiled using ssGSEA, CIBERSORT, and ESTIMATE, while immunophenoscore (IPS) was used to predict immunotherapy responsiveness. Functional enrichment, CellChat, SCISSOR, tumor mutation burden (TMB), and CellMiner analyses were further performed to explore underlying mechanisms and therapeutic implications. A three-gene signature (CD9, NPC2, PSMB9) effectively stratified patients into highand low-risk groups with distinct survival outcomes. Low-risk tumors exhibited an "immune-hot" phenotype with increased CD8+ T cells and activated NK cells, higher checkpoint expression, and elevated IPS, suggesting greater immunotherapy sensitivity. In contrast, high-risk tumors showed an immune-cold microenvironment with M2 macrophage enrichment. Despite higher TMB, high-risk tumors displayed reduced immune activity, indicating impaired immune recognition. Single-cell analysis further identified MIF and CCL signaling as key mediators of multicellular immune evasion. Overall, our single-cell-informed immune escape signature provides a promising framework for thyroid carcinoma risk stratification and offers insights into personalized immunotherapy.
Passive leg raising (PLR) is a common maneuver to assess fluid responsiveness in the emergency department. The effect of blood pressure (BP) on PLR-induced left ventricular (LV) stroke volume (SV) response was examined in normotensive (CON, n = 100), hypotensive (HOTN, n = 51) and hypertensive (HTN, n = 16) individuals. Limb BP and Echo measurements were completed within 1 min when the participants were transitioned from the semi-recumbent position to bilateral leg elevation at 45° and lower limb BP was reduced at PLR in all three groups. LV end-diastolic diameter (LVDD), LV end-systolic diameter (LVDS), end-systolic or end-diastolic volume (ESV, EDV) were comparable at baseline among three groups, whereas E, E/A ratio, and deceleration time were significantly lower and A was significantly greater in the HOTN and HTN groups compared with the CON group (p < 0.05). Notably, EDV, LVDD and SV, cardiac output, ejection fraction, and fractional shortening were increased significantly after PLR only in HOTN group (p < 0.05). EDV and SV were positively correlated with E and the E/A ratio in HOTN (p < 0.05), whereas a negative correlation was observed in CON group (p < 0.05). Receiver operating characteristic (ROC) analysis demonstrated that baseline E < 0.77 m/s and systolic BP < 101 mmHg effectively predicted an SV increase of > 15% at PLR with good sensitivity and specificity. Mean arterial pressure, diastolic BP and E wave velocity confine SV responsiveness to PLR even in CON individuals. These findings indicate inherent physiological thresholds for SV augmentation via the Frank-Starling mechanism even in the general population.
Jiajian Xiefei Tang (JJXFT) is a practical prescription rooted in traditional Chinese medicine clinical experiences for various pulmonary diseases. Nevertheless, its function and mechanism in asthma treatment remain unclear. An ovalbumin-induced murine asthma model was established. The effect of JJXFT on airway inflammation was assessed via histopathological examination and ELISA. Network pharmacology analysis was conducted to predict potential targets and upstream active compounds of JJXFT, followed by validation of binding interactions via molecular docking and cellular thermal shift assay. Western blot and immunofluorescence were applied to detect proteins linked to the STAT3 signaling axis. An in vitro alveolar epithelial cell injury model was built to further explore the molecular mechanism of JJXFT in ameliorating asthma. JJXFT alleviated disease progression in asthmatic mice by improving pathological conditions and airway inflammation in lung tissues. Network pharmacology analysis detected STAT3 as a potential target of JJXFT, and licochalcone A (Lico-A) as an active component. Experimental validation confirmed that Lico-A bound to STAT3, markedly suppressing its phosphorylation and nuclear translocation. In vitro studies demonstrated that Lico-A effectively reduced asthma-related cellular damage by inhibiting the STAT3/IL-17 signaling pathway. JJXFT attenuates asthma symptoms by modulating the STAT3/IL-17 signaling axis through its active component Lico-A, suggesting that JJXFT may function as a potential therapeutic agent for asthma.
Pediatric septic shock is a severe form of sepsis with high mortality. Histone deacetylation is involved in sepsis-related disorders. This study investigated the diagnostic value of histone deacetylation-related genes in pediatric septic shock. Three Gene Expression Omnibus datasets (GSE26378, GSE26440, and GSE13904) were analyzed. Differentially expressed genes from GSE26378 and GSE26440 were intersected with genes in key modules identified by weighted gene co-expression network analysis. Least absolute shrinkage and selection operator regression and random forest were used to identify hub genes, and receiver operating characteristic curves assessed diagnostic performance. Quantitative real-time polymerase chain reaction validated candidate gene expression. Immunoprecipitation preliminarily evaluated protein acetylation changes after trichostatin A treatment. Immune infiltration was analyzed, and related transcription factors and miRNAs were predicted. C9orf84, ASAP1-IT1, and CREB5 were identified as hub genes. All were upregulated in pediatric septic shock and showed favorable diagnostic performance in the training and validation datasets. Trichostatin A increased the acetylation levels and reduced the protein levels of C9orf84 and CREB5. Neutrophils, macrophages, and regulatory T cells were more abundant in pediatric septic shock than in controls. Multiple potentially interacting transcription factors and miRNAs were also identified. C9orf84, ASAP1-IT1, and CREB5 may serve as diagnostic biomarkers and therapeutic targets for pediatric septic shock.
Type 1 diabetes mellitus is characterized by insulin deficiency, hyperglycemia, and systemic metabolic and inflammatory disturbances that lead to multi-organ injury. Building on prior evidence that 6'-sialyllactose (SL), a sialylated human milk oligosaccharide, exerts anti-inflammatory and metabolic benefits, we investigated whether SL mitigates streptozotocin (STZ)-induced metabolic dysfunction and tissue injury in vivo. Male ICR mice were pretreated with SL (25 or 75 mg/kg) prior to STZ administration. STZ challenge markedly increased serum glucose levels, disrupted hepatic enzyme profiles, impaired protein metabolism, and induced liver apoptosis and inflammation. SL pretreatment significantly improved glucose homeostasis, normalized liver enzyme activities and serum protein indices, reduced hepatic caspase-3 activation, and suppressed pro-inflammatory cytokine expression in the liver and skeletal muscle. Mechanistically, SL restored AMPK-Akt-mTOR signaling in metabolic tissues. Collectively, these data demonstrate that SL confers systemic metabolic protection against STZ-induced diabetes, highlighting its potential as a therapeutic or preventive agent targeting inflammatory and metabolic pathways.
Oxaliplatin (OXA) resistance in colorectal cancer (CRC) remains a major therapeutic challenge, prompting investigation into the role of the interferon regulatory factor 4 (IRF4)/leukocyte differentiation antigen 36 (CD36) regulatory axis and its impact on glycolysis. Bioinformatic analysis identified CD36 as a key gene, with IRF4 predicted as its upstream regulator. OXA-resistant CRC cell lines were established. Gene expression was analyzed by reverse transcription quantitative polymerase chain reaction and Western blot. The IRF4-CD36 interaction was validated via chromatin immunoprecipitation and dual-luciferase assays. Glycolysis was assessed by measuring glucose consumption, lactate production, and ATP levels. Drug sensitivity was evaluated using cell counting kit-8 and apoptosis assays. Both IRF4 and CD36 were downregulated in OXA-resistant cells. IRF4 directly bound to the CD36 promoter to activate its transcription. Overexpression of CD36 or IRF4 inhibited glycolysis and restored OXA sensitivity. Knockdown of CD36 in IRF4-overexpressing cells reversed these effects, confirming CD36 as the functional mediator. The study demonstrates that the IRF4/CD36 axis attenuates OXA resistance in CRC by transcriptional repression of glycolysis. This axis presents a promising therapeutic target for overcoming chemoresistance.
Bone morphogenetic protein-2 (BMP-2) regulates cell differentiation and proliferation. However, its role in colorectal cancer (CRC) remains debatable owing to potential oncogenic effects. Here, we investigated the role of BMP-2 in modulating CRC progression, particularly in regulating the Hippo signaling pathway. Exposure to recombinant human BMP-2 (rhBMP-2) resulted in a concentration-dependent decrease in CRC cell proliferation, leading to G1 cell cycle arrest. This effect was associated with the increased expression of p53, p21, and Smad4, while the levels of cyclin D1, cyclin-dependent kinase 4 (CDK4), and CDK6 decreased. Additionally, rhBMP-2 promoted apoptosis by decreasing poly (ADP-ribose) polymerase and caspase-9 expression while increasing their cleaved forms. It also activated the Hippo signaling cascade, enhancing the expression of mammalian sterile 20-like kinase 1/2, Mps One Binder 1 (MOB1), phosphorylated MOB1, and Salvador homolog, along with elevated levels of phosphorylated yes-associated protein (YAP), while concurrently suppressing total YAP expression. This resulted in cytoplasmic sequestration and subsequent degradation of YAP, thereby attenuating the transcription of YAP-responsive genes such as Connective Tissue Growth Factor. Silencing of Ras association domain family member 1 restored the rhBMP-2-induced decrease in cell viability, whereas silencing YAP further reduced the viability of CRC cell lines. Administering rhBMP-2 significantly suppressed tumor expansion in a mouse model of CRC, further supporting its potential as an antitumor agent. Collectively, these results indicate that rhBMP-2 mitigates CRC progression by activating the Hippo signaling pathway and suppressing YAP-mediated oncogenic processes, thereby highlighting its potential as a therapeutic agent that warrants further clinical evaluation.
Sweating is the primary mechanism for human thermoregulation, driven by eccrine gland activity under central and peripheral sudomotor control. In this review, we synthesize evidence showing that sudomotor function is shaped by biological traits (age, sex, ethnicity/ancestry), environmental exposures (regional climate, seasonality, and migration), and individual modifiers (exercise training and dermatologic disease). Environmental heat exposure induces heat adaptation, leading to short-term and long-term acclimation with contrasting effects. Short-term acclimation enhances sweat gland sensitivity and density, increasing sweat output for efficient cooling during heat stress. Conversely, long-term acclimation suppresses sweating to conserve body fluids by reducing sweat gland responsiveness and output. Regional and seasonal differences in sweating patterns are closely linked to climate adaptations. Age-related declines in sweat gland function are evident; children exhibit higher sweat gland density but lower output per gland due to immature sudomotor function. Exercise training improves sweat gland sensitivity and density, enhancing thermoregulation during physical activity. Dermatologic conditions such as atopic dermatitis and psoriasis impair sweat secretion through skin barrier dysfunction and autonomic dysregulation, exacerbating inflammation and compromising thermoregulatory efficiency. This review highlights the complex interplay between intrinsic biological traits, external environmental stimuli, and individual behaviors in shaping sudomotor function. Understanding these mechanisms can inform strategies for optimizing thermoregulation and managing sweat-related impairments in clinical settings.
Protocatechuic acid (PCA) and tanshinone IIA (TanIIA), the major functional constituents of Phellinus baumii (PB) and Salvia miltiorrhiza (SM), respectively, regulate innate and adaptive immune pathways and converge on nuclear factor-κB (NF-κB) signaling, providing a mechanistic basis for cooperative immune enhancement. Based on these characteristics, this study evaluated the immunomodulatory effects of a PB-SM complex and examined whether combined treatment with PCA and TanIIA could cooperatively enhance immune recovery. In a cyclophosphamide (CPA)-induced immunosuppressed mouse model, PB-SM administration significantly alleviated body weight loss and splenic atrophy and restored the reduced proportions of CD3+ T cells, CD335+/NKR-P1C+ NK cells, and F4/80+ macrophages. PB-SM also significantly improved the proliferative responses of T and B cells suppressed by CPA. At the cellular level, PB-SM enhanced phagocytic activity of macrophages and NK cell-mediated tumor killing. Consistently, PCA and TanIIA individually activated MAPK and NF-κB p65 signaling and improved macrophage functions, with the combination exhibiting enhanced activity. These findings suggest that PCA and TanIIA may contribute to immune restoration and support their combined application as a promising functional strategy for alleviating chemotherapy-induced immunosuppression.
Homocysteine (HCy) is a sulfur-containing metabolic intermediate with largely unexplored potential as a damage-associated molecular pattern. This study investigated the immunomodulatory profile of HCy alone and in combination with the TLR4 agonist monophosphoryl lipid A (MPL). To model acute metabolic stress, we evaluated murine bone marrow-derived dendritic cells (DCs), macrophages, and splenocytes treated with HCy (6.25-600 μg/ml) in vitro, and assessed inflammatory cell recruitment in vivo in mice injected intraperitoneally with HCy (100 or 500 μg/mouse). HCy treatment significantly increased mitochondrial metabolic activity (CCK-8) without generalized necrosis. However, high concentrations (400-600 μg/ml) paradoxically suppressed MPL-induced splenocyte proliferation. HCy displayed distinct cell-specificity, acting as a direct agonist for macrophage maturation (upregulating CD40, CD86, and MHC-II) while failing to activate purified DCs. Notably, flow cytometry revealed that while metabolic signals increased, high-dose HCy induced late apoptosis in macrophages starting at 25 μg/ml, suggesting a state of metabolic stress rather than enhanced viability. Crucially, HCy exerted a regulatory effect on TLR4 signaling: co-treatment with HCy (200-600 μg/ml) dose-dependently suppressed MPL-induced pro-inflammatory cytokine secretion (TNF-α, IL-6) while maintaining phenotypic maturation. In vivo, HCy functioned as a chemotactic agent, recruiting DCs and monocytes to the peritoneal cavity and inducing an activated, M2-like (CD206+) macrophage phenotype. These findings demonstrate that HCy functions as a dual-action immune modulator that promotes phenotypic maturation and antigen-presenting cell recruitment while dampening excessive cytokine release through a stress-mediated regulatory mechanism.
Hepatocellular carcinoma (HCC) metastasis often threatens patient survival. However, there is still a lack of effective treatments to tackle this problem. Ruanjianhugan tablets (hepatoprotective and fibrolytic tablets), a traditional Chinese herbal compound, have demonstrated potential in repressing the progression of HCC. However, the exact active ingredients and mechanisms behind its effects are not yet fully understood. In this study, we leveraged network-pharmacology tools to screen the key bioactive constituent-calycosin (Cal)-from this compound formula and predicted it as the target for the downstream gene checkpoint kinase 1 (CHEK1) in HCC cells. Subsequent cellular thermal shift assay (CETSA) verified that Cal bound CHEK1. In vitro, CCK-8, colony-formation, Transwell and Western-blot assays showed that Cal markedly suppressed HCC-cell proliferation, colony formation, migration, invasion and epithelial-to-mesenchymal transition. In xenograft nude-mouse and tail-vein lung metastasis models, exogenous Cal bound CHEK1 and thereby inhibited HCC metastasis and tumor growth. Collectively, these results not only establish Cal as the core pharmacodynamic component of Ruanjianhugan tablets but also unveil a novel Cal-CHEK1 axis underlying anti-HCC metastasis.
Binaphthoquinone (BiNQ), a dimeric compound composed of juglone (5-hydroxy-1,4-naphthoquinone) and naphthazarin (5,8-dihydroxy-1,4-naphthoquinone), was originally isolated from the sea urchin Spatangus purpureus O.F. Müller. In this study, we examined the anti-fibrotic effects of BiNQ in a diabetic context, focusing on its potential to enhance mitochondrial function. Human cardiac fibroblasts (NHCF-V) were cultured under control 5 mM glucose or exposed to 33 mM glucose plus palmitic acid (100 μM). Fibroblast-to-myofibroblast transition was evaluated by measuring the expression level of the myofibroblast marker α-smooth muscle actin (α-SMA), and functional changes were further assessed using a contraction assay. BiNQ was applied at two concentrations (10 nM and 1 μM) and was found to reduce α-SMA expression, indicating inhibition of myofibroblast differentiation. Concurrently, BiNQ enhanced the expression of mitochondrial respiratory chain complexes, resulting in increased ATP production. To elucidate the underlying molecular mechanisms, targeted proteomic analysis was performed using LC-MS/MS, enabling identification and quantification of proteins involved in mitochondrial bioenergetics and fibrotic regulation. Collectively, these findings suggest that BiNQ attenuates fibroblast activation in a diabetic context by improving mitochondrial function and modulating key pro-fibrotic pathways.
Polycystic ovary syndrome (PCOS) affects 11%-13% of reproductive-age women worldwide and is pathologically associated with granulosa cell dysfunction. This study employed transcriptome sequencing of granulosa cells from PCOS patients and non-PCOS controls, followed by Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment Analysis. Key differentially expressed genes were validated by quantitative real-time PCR. Transcriptome analysis identified 157 upregulated and 71 downregulated mRNAs in PCOS granulosa cells, with enrichment in PI3K-Akt, MAPK, and TGF-beta signaling pathways. Six genes-NPTX2, FN1, CCN1, IDH1, ZCCHC17, and CREG1-were confirmed by qRT-PCR. Protein-protein interaction network analysis identified FN1 as a central hub gene. FN1 knockdown in KGN cells suppressed proliferation, induced apoptosis, and caused G1 phase arrest, accompanied by reduced Akt phosphorylation and altered expression of cyclin D1, p21, and p27. These findings suggest a potential association between FN1 and granulosa cell proliferative dysregulation in PCOS, warranting validation in primary cells and in vivo models.
Osteoporosis, characterized by excessive bone resorption, requires new therapeutic agents with high efficacy and favorable safety profiles. In this study, we isolated alpha-mangostin (aMG) from the peel of Garcinia mangostana L. and investigated its osteogenic, antiresorptive, and toxicological activities. The osteogenic effects were evaluated in vitro using MC3T3-E1 pre-osteoblast cells, whereas acute toxicity and bone-protective activity were assessed in vivo using medaka fish (Oryzias latipes) . In MC3T3-E1 cells, aMG promoted osteogenic differentiation by enhancing proliferation, alkaline phosphatase activity, collagen synthesis, and mineralization at 1 and 10 μM, whereas cytotoxicity was observed at 100 and 1,000 μM. In medaka embryos and larvae, aMG induced acute toxicity with LC50 values (concentrations causing 50% mortality) of 91.27 and 3.44 μM, and EC50 values (concentrations causing 50% of developmental abnormalities) of 103.66 and 3.3 μM, respectively, leading to tail deformities, hemorrhage, and reduced hatching and heart rates. At sub-toxic concentrations (0.25-1 μM), aMG significantly attenuated Rankl-induced bone loss in the neural arches of osteoporotic medaka larvae. Collectively, this study provides the first evidence of the dual osteogenic and antiresorptive actions of aMG, along with its toxicity profile, highlighting its potential as a plant-derived anti-osteoporosis agent.
Ischemic stroke, one of the world's leading fatal diseases, has a high incidence and recurrence, leading to severe mortality and disability. In this study, we investigated whether treadmill exercise is an important treatment to prevent recurrence and improve functional impairment following an ischemic stroke. Experimental cerebral ischemia by occluding the middle cerebral artery was induced in rats, and the effect of 10- or 30-min training for two weeks was evaluated. To assess for motor function improvement, behavioral tests including the elevated body swing test were conducted. The expressions of the endoplasmic reticulum (ER) stress and apoptosis markers were investigated by Western blotting analysis and immunohistochemistry. In both exercise groups (10 and 30 min), motor function improved compared to the non-exercise group. TTC staining demonstrated that the brain infarct volume also decreased after exercise. Further examination of the signaling pathway showed that the expression of ER stress-related proteins, such as IRE1-α, JNK, ERK, and p38 MAPK, decreased significantly in the exercise groups. The pro-apoptotic genes (Bax and pro-caspase3) of the apoptosis signaling mechanism, also decreased in the exercise groups. Interestingly, the level of neuronal markers (NeuN, SYP, and NFH) increased in the exercise groups. Our results suggest that exercise has a beneficial effect following ischemic stroke. In particular, exercise used in short- or long-term training could regulate the signaling mechanisms, such as ER stress, apoptosis, and neuronal cell death protection.
Curcumin (CUR) is used in treating knee osteoarthritis (KOA), but its effects on synovial fibrosis and underlying mechanisms remain unclear. In vivo, a rat KOA model was established via anterior cruciate ligament transection (ACLT), followed by CUR administration. Synovial fibrosis, autophagy, and PI3K/AKT/mTOR pathway were assessed. In vitro, TGF-β1-induced fibroblast-like synoviocytes (FLSs) were treated with CUR. Fibrosis markers, autophagy activity, and PI3K/AKT/mTOR pathway proteins were analyzed. CUR alleviated synovial fibrosis in ACLT-induced rats. In FLSs, CUR reduced TGF-β1-stimulated fibrosis, suppressed PI3K/Akt/mTOR signaling, and enhanced autophagy. In vivo results confirmed CUR inhibited PI3K/Akt/mTOR and activated autophagy. CUR attenuated synovial fibrosis by activating protective autophagy via inhibition of the PI3K/Akt/mTOR pathway, elucidating a novel anti-fibrotic mechanism for KOA therapy.
Asthma is a significant allergic condition affecting the respiratory system. Numerous compounds extracted from traditional Chinese herbal medicine show potential benefits for treating airway inflammation associated with bronchial asthma. Astragalus polysaccharide (APS), a class of major extracts from Astragalus membranaceus, exhibit many anti-inflammatory effects. Nonetheless, the underlying mechanisms of APS in asthma remain to be clarified. This study aims to assess the effect and the mechanism by which APS against asthma. The ovalbumin (OVA)-induced asthma mice were employed to assess the roles of APS in vivo. Lung tissues were used for H%E, PAS, and Masson staining. Inflammatory cells in bronchoalveolar lavage fluid (BALF) and chemokines in both BALF and serum samples were determined by hemocytometer and ELISA. The expression of the Hedgehog/NLRP3/GSDMD pathway in lung tissues was measured by quantitative real-time PCR and Western blotting. APS displayed lung-protective effects through decreasing airway inflammation and airway remodeling in OVA-induced asthma mice, which was demonstrated by decreasing the number of inflammatory cells and the cytokine levels in BALF and serum, and histopathological changes in lung tissues. Besides, APS treatment could decrease the expression of SHH, SMO, Gli1, Ptch1, NLRP3, GSDMD-N, ASC, Cleaved caspase-1, IL-18, IL-1β, CARMA3, BCL10, and MALT1 in lung tissues. APS improves clinical symptoms of OVA-induced bronchial asthma. The anti-asthmatic effects of APS may be related to the regulatory influences of the Hedgehog/NLRP3/GSDMD pathway.
