Background To investigate the molecular mechanism by which deoxyshikonin (DS) modulates hsa-HLA-DRB1 to ameliorate high glucose (HG)-induced dysfunction in human umbilical vein endothelial cells (HUVECs) and to evaluate its regulatory effect on wound healing in a diabetic mouse model, thereby providing an experimental basis and a targeted therapeutic strategy for diabetic foot ulcers (DFU). Methods An in vitro model was established using HUVECs subjected to HG and hsa-HLA-DRB1 overexpression with or without DS intervention. Differentially expressed genes (DEGs) were identified via RNA sequencing (RNA-seq). Cell proliferation, migration, tube formation, and apoptosis were assessed using CCK-8, wound healing, tube formation assays, and flow cytometry, respectively. Key pathway molecules and inflammatory cytokines were quantified using real-time quantitative PCR (RT-qPCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Dual-luciferase reporter assays were performed to validate the targeting relationships between hsa-HLA-DRB1 and miRNA-12118, and between miRNA-12118 and FLT-1. An in vivo model was established using C57BL/6 diabetic mice with dorsal skin wounds. Wounds were locally injected with lentivirus overexpressing hsa-HLA-DRB1. Mice were randomized into five groups: normal control, diabetes mellitus (DM), DM + DS, DM + hsa-HLA-DRB1 lentivirus (OE-hsa-HLA-DRB1), and DM + OE-hsa-HLA-DRB1 + DS. Wound healing rates were calculated, and molecular markers were analyzed. Results RNA-seq analysis identified 2,415 DEGs across the four cell groups, with significant enrichment in pathways related to immune inflammation, signal transduction, and angiogenesis. Dual-luciferase assays confirmed direct binding between hsa-HLA-DRB1 and miRNA-12118, and between miRNA-12118 and FLT-1. Overexpression of hsa-HLA-DRB1 under HG conditions significantly inhibited HUVEC migration, proliferation, and tube formation (P < 0.05), enhanced the inflammatory response (P < 0.05), but had no significant effect on apoptosis. Intervention with 3 µmol/L DS reversed these cellular phenotypes, downregulated the expression of molecules such as CXCL12 (P < 0.05), and decreased the concentrations of IL-6 and TNF-α (P < 0.001). In vivo, the wound healing rate in the DM + OE-hsa-HLA-DRB1 group was significantly lower than that in the DM group (P < 0.01), with high hsa-HLA-DRB1 expression and a markedly enhanced inflammatory response in wound tissue. DS treatment significantly increased the wound healing rate in both the DM and DM + OE-hsa-HLA-DRB1 groups. In vivo, the normal control group showed a 14-day wound healing rate of 95.2 ± 2.35%. The DM group exhibited delayed healing (87.4 ± 3.86%, P < 0.001 vs. control). The DM + OE-hsa-HLA-DRB1 group had significantly impaired healing (85.4 ± 4.12%, P < 0.01 vs. DM). DS treatment markedly improved healing: the DM + DS group reached 92.5 ± 3.26% (P < 0.001 vs. DM), and the DM + OE-hsa-HLA-DRB1 + DS group achieved 93.3 ± 3.58% (P < 0.001 vs. DM + OE-hsa-HLA-DRB1). RT-qPCR and Western blot analyses confirmed that DS inhibited the activation of the PI3K/Akt and TNF inflammatory pathways, alleviated oxidative stress injury, and promoted granulation tissue formation, angiogenesis, and re-epithelialization. Conclusions DS targets the hsa-HLA-DRB1, effectively ameliorating HG-induced HUVEC dysfunction in vitro and reversing hsa-HLA-DRB1 overexpression-mediated delayed wound healing in diabetic mice in vivo. These findings identify DS as a promising therapeutic candidate and hsa-HLA-DRB1 as a potential target for DFU treatment.

BACKGROUND:Intracerebral hemorrhage (ICH) induces severe neuroinflammation and microglial pyroptosis, exacerbating secondary brain injury. Rhein, a natural anthraquinone compound, possesses anti-inflammatory and neuroprotective properties. However, its effects on microglial pyroptosis and the underlying mechanisms remain unclear. METHODS:A rat microglial (RM) pyroptosis model was established using LPS + ATP induction, followed by rhein intervention and NLRP3 knockdown. Cell proliferation was assessed using CCK-8, and apoptosis was evaluated through TUNEL staining. ELISA was used to measure the expression levels of inflammatory cytokine. Immunofluorescence staining was performed to label M1/M2 microglia. RT-qPCR and western blot were used to analyze the expression of NLRP3, ASC, Caspase-1, GSDMD, PCNA, Cyclin D1, and CDK2. Transmission electron microscopy (TEM) was used to observe pyroptotic bodies. Additionally, a rat ICH model was established, with rhein intervention and NLRP3 knockdown/Caspase-1 inhibition. Behavioral assessments were conducted using the Y-maze test and open-field test. HE staining was performed to examine brain tissue pathology. ELISA was used to measure inflammatory cytokine levels in brain tissue. Immunofluorescence staining analyzed the distribution of M1/M2 microglia. RT-qPCR and western blot were used to detect pyroptosis-related proteins, and TEM was used to evaluate pyroptotic body formation. RESULTS:At the cellular level, rhein significantly promoted microglial proliferation and M2 polarization while inhibiting pyroptosis, inflammatory cytokine expression, M1 polarization, and NLRP3 expression. NLRP3 knockdown further enhanced the protective effects of rhein. At the animal level, ICH model rats exhibited reduced exploratory behavior, exacerbated neuroinflammation, increased pro-inflammatory cytokine expression, increased M1 microglia, and elevated NLRP3 expression and pyroptosis levels. Rhein intervention significantly alleviated inflammation in ICH rats by reducing the expression of NLRP3 and pyroptosis-related proteins. NLRP3 knockdown or Caspase-1 inhibition further enhanced rhein's protective effects. CONCLUSION:Rhein alleviates neurological dysfunction following ICH by inhibiting NLRP3 inflammasome activation, reducing microglial pyroptosis, and mitigating neuroinflammation.

Background: This study investigated the role of miR-23a-3p in the pathogenesis of tuberculosis (TB)-associated obstructive pulmonary disease (TOPD) and its regulatory impact on THP-1 macrophages via the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway. Methods: Patients with TOPD, TB patients without airflow obstruction (only TB), and healthy controls were recruited. Peripheral blood mononuclear cells were isolated, followed by RNA-seq analysis to identify differentially expressed microRNAs. Gene set enrichment analysis was used to identify enriched biological pathways. Additionally, enzyme-linked immunosorbent assays, Western blot, and flow cytometry were used to explore the miR-23a-3p-mediated modulation in macrophages (such as apoptosis and polarization) via the JAK-STAT pathway. Results: RNA-seq analysis identified miR-23a-3p as being significantly upregulated in TOPD patients. Bioinformatics analysis indicated that miR-23a-3p targets regulation of the JAK-STAT pathway. Overexpression of miR-23a-3p in macrophages led to decreased JAK1 protein expression and reduced levels of phosphorylated JAK1 and STAT1. Functional assays revealed that miR-23a3p mitigates macrophage apoptosis and macrophage polarization as well as influences inflammatory cytokine production. Conclusion: miR-23a-3p regulates TOPD pathogenesis by modulating macrophage inflammation, apoptosis, and differentiation via the JAK-STAT pathway, making it a promising immunotherapeutic target for future treatments.

Mitochondrial ribosomal protein L13 (MRPL13) has been implicated in tumor progression, but its relevance to esophageal squamous cell carcinoma (ESCC) remains insufficiently defined. Because mitochondrial adaptation can influence tumor cell fitness under oncogenic and oxidative stress, we investigated whether MRPL13 is associated with aggressive ESCC biology and mitochondrial functional states. We analyzed MRPL13 expression, clinical associations, survival relevance, mutation patterns, immune-related correlates, and pathway enrichment using TCGA and related public datasets, with an emphasis on ESCC-specific analyses. Experimental validation was performed using paired ESCC tissues and KYSE150 cells with shRNA-mediated MRPL13 knockdown. Western blotting, colony formation, transwell migration/invasion assays, mitochondrial membrane potential assessment, and ROS staining were used to evaluate protein expression, cellular behavior, and mitochondrial status. MRPL13 was up-regulated in ESCC and its expression was associated with tumor stage and unfavorable prognosis. MRPL13 expression correlated with a set of cancer-associated genes, including SIX2, SPP1, COL11A1, DSC3, and CCNA1, several of which were also increased at the protein level in paired ESCC tissues. ESCC tumors with higher MRPL13 expression showed distinct mutation and immune-related expression patterns; however, these immune associations were interpreted as correlates of tumor state rather than direct evidence of immune regulation. Pathway analyses indicated enrichment of MAPK-related signaling in MRPL13-high tumors. Consistently, MRPL13 knockdown in KYSE150 cells reduced phosphorylation of MEK1/2, ERK1/2, JNK1/2, and p38, impaired colony formation and in vitro migration/invasion, decreased mitochondrial membrane potential, and increased intracellular ROS levels. These findings suggest that MRPL13 identifies an aggressive ESCC state in which mitochondrial integrity, MAPK-associated signaling, and tumor cell fitness are functionally coupled. Rather than establishing MRPL13 as a fully defined upstream driver, our data support a more restrained interpretation: MRPL13 may help ESCC cells maintain mitochondrial stress tolerance, and its loss exposes a vulnerability characterized by reduced MAPK phosphorylation, impaired clonogenic and invasive capacity, mitochondrial depolarization, and ROS accumulation.

Objective:This study aimed to investigate the protective effects of hordenine on concanavalin A (ConA)-induced immune liver injury (ILI). Methods:Liver coefficients and spleen coefficients were determined. The serum levels of alanine transaminase (ALT) and aspartate transaminase (AST) were detected. Hematoxylin-eosin (H&E) staining was used to observe the pathological changes of the liver, spleen, and thymus. Immunohistochemical experiments were performed to detect the expression of inflammatory factors interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) in liver tissues. Superoxide dismutase (SOD) and malondialdehyde (MDA) contents were detected. TdT-Mediated dUTP Nick-End Labeling (TUNEL) staining was used to observe the apoptosis of liver cells. Western Blot was used to detect the expression of B-cell lymphoma-2 (Bcl-2), Caspase-3, Bcl-2-Associated X (Bax), kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1). Results:We found that hordenine alleviated liver and spleen enlargement, ameliorated pathological damage in the liver, spleen, and thymus, and reduced transaminase and inflammatory factor levels in the liver. Moreover, hordenine increased the level of SOD while decreasing the level of MDA, attenuated hepatic apoptosis, decreased the expression of caspase-3, Bax, and Keap1 and increased the expression of Bcl-2, Nrf2, and HO-1. Conclusion:These results suggest that hordenine has a protective effect against ConA-induced immune liver injury, which may be related to its modulation of the Keap1/Nrf2/HO-1 pathway and inhibition of oxidative stress and apoptosis.

ObjectiveTo elucidate the metabolic mechanisms by which acteoside (ACT) isolated from Cistanche tubulosa alleviates cancer-related fatigue (CRF) in a murine model of colon cancer with cachexia.MethodsBALB/c mice inoculated with C26 colon cancer cells were treated with paclitaxel (PTX, 10 mg/kg) and ACT (100 mg/kg) alone or in combination for 21 days. Fatigue-associated behaviors, tumor inhibition rate, and skeletal muscle morphology assessed by hematoxylin-eosin (H&E) staining and electron microscopy were evaluated. Finally, liquid chromatography-mass spectrometry (LC/MS) was employed to investigate alterations in the plasma metabolic profile of tumor-bearing mice with CRF in response to ACT treatment, and the affinity between metabolite-associated proteins and ACT was verified by Surface plasmon resonance (SPR) assay.ResultsOur study demonstrated the presence of CRF in the colon cancer mouse model, with the severity of fatigue increasing alongside tumor growth. Administration of ACT ameliorated both tumor burden and PTX-induced muscle fatigue-like behavior. LC/MS analysis identified a panel of differentially regulated metabolites, including trans-aconitine, citric acid, 3-coumaric acid, ephedrine, thymine, cytosine, indole-3-acetic acid, and pantothenol-9. These metabolites were primarily enriched in pathways associated with valine biosynthesis, tyrosine metabolism, tryptophan metabolism, and biosynthesis of pyridine alkaloids. Furthermore, several key enzymes, including CYP3A4, CYP19A1, CYP2E1, TNF, BCL-2, RYR2, and ATP2A1, were identified as potential targets underlying the anti-CRF effects of ACT.ConclusionThis study suggests that ACT derived from C. tubulosa harbors protective properties against cancer-related fatigue mediated by tumor cells.

Cancer cells evade immune detection through checkpoint molecules like PD-L1 and PD-L2 which suppress T-cell activation. While PD-L1 is well-studied, the role of PD-L2 remains unclear. Pyruvate kinase M2 (PKM2), a metabolic enzyme, influences immune checkpoint regulation, but its role in PD-L1 and PD-L2 modulation is not well defined. Here, we investigate the role of pyruvate kinase M2 (PKM2) in modulating the immune checkpoint molecules PD-L1 and PD-L2 via GATA3 in cancer cells, with insights from both human and mouse models. We find that PKM2 enhances PD-L1 expression while inhibiting PD-L2, a dual regulatory mechanism that facilitates immune evasion. Knockdown and overexpression experiments revealed GATA3 as a key mediator. PKM2 knockout reduced GATA3 level, leading to decreased PD-L1 and increased PD-L2 expression. Chromatin immunoprecipitation (ChIP)-qPCR demonstrates that GATA3 functions as a direct transcription factor capable of binding to the promoters of PD-L1 and PD-L2. In silico analyses of 81 esophageal squamous cell carcinoma (ESCC) cases from the TCGA database demonstrate that PKM2 mRNA is unrelated to PD-L1 and PD-L2 expression but is negatively correlated with CD8 + T-cell infiltration in ESCC. To further validate these findings, we establish a xenograft model using immune-competent C57/BL6N mice, where knockdown of PKM2 results in significant downregulation of both PD-L1 and PD-L2 expression. Collectively, these findings underscore the divergent roles of PKM2 in regulating immune checkpoint expression in human and mouse cancer models and suggest that targeting the PKM2-GATA3 axis could enhance cancer immunotherapy by fine-tuning PD-L1 and PD-L2 levels.

Nervous necrosis virus poses a significant challenge to the aquaculture industry, resulting in substantial financial losses globally. In this study, we evaluated the anti-NNV activity of luteolin, a flavonoid natural product. The results demonstrated that luteolin, at a concentration of 27.95 mu M, significantly inhibited NNV-induced cytopathic effects. Additionally, the half-inhibitory concentration for luteolin against NNV replication was determined to be 9.87 mu M. Importantly, luteolin was shown to inhibit NNV proliferation by suppressing NNV-induced late apoptosis. Furthermore, luteolin demonstrated a preventive effect against NNV infection. To enhance the in vivo antiviral efficacy of luteolin and address the selective permeability of the blood-brain barrier (BBB), we prepared a luteolin-borneol complex. The presence of borneol significantly enhanced the in vivo antiviral activity of luteolin, likely due to its ability to improve BBB permeability, thereby enhancing luteolin's bioavailability. These findings suggested that luteolin is a potent anti-NNV agent, and that the luteolin-borneol complex represented a novel therapeutic approach with promising applications for combating NNV infections.

BackgroundThe tumor microenvironment (TME) is crucial in influencing the progression and therapeutic response of ovarian cancer.MethodThis study conducted a comprehensive bibliometric and visualization analysis of research trends and focal areas concerning the ovarian cancer microenvironment from 2005 to 2024. A total of 1,720 pertinent articles were identified from the Web of Science Core Collection (WoSCC) database.ResultsThe analysis revealed a notable increase in research activity on the TME of ovarian cancer, particularly during the period from 2019 to 2022. The United States and China were the most active countries in this field, and the University of Texas System was the scientific research institution with the largest number of articles. Cancer Research and Clinical Cancer Research were the most cited journals. Weiping Zou and Anil K Sood were recognized as the most cited researchers. The study systematically identified key research hotspots within this field, encompassing immune checkpoint inhibitors, single-cell RNA sequencing technology, the TME heterogeneity, the TGFβ signaling pathway, and the impact of PARP inhibitors on the tumor immune microenvironment (TIME).ConclusionThis study provides a valuable reference for the evolution and prospective directions of TME research in ovarian cancer, underscoring the critical importance of a comprehensive understanding of the TME to enhance treatment strategies for ovarian cancer.

We investigated the role and mechanism of hsa-HLA-DRB1 in the development and progression of diabetic foot ulcers. High-throughput sequencing was performed on three normal foot trauma tissues and diabetic foot ulcer tissues. The circRNAs with significant differences were identified. The downstream miRNAs were predicted by miRanda and RNAhybrid databases, and the mRNAs were predicted by the TargetScan database. Validation was performed with CCK8, flow cytometry, trabecular scratch assay, tubule generation assay, Western blot, dual luciferase assay, and RT-qPCR. High-throughput sequencing identified 461 significantly different circRNAs, of which 260 were up-regulated and 201 down-regulated. Compared to normal tissue, hsa-HLA-DRB1 was highly expressed in diabetic foot ulcers. The hsa-HLA-DRB1/miRNA_12118/FLT-1 axis was constructed. In vitro, we found that HLA-DRB1 overexpression inhibited cell viability, wound healing, and tubule formation, promoted apoptosis, and enhanced FLT-1 expression in HUVECs. The upregulation of hsa-HLA-DRB1 may promote diabetic foot development The upregulation of hsa - HLA - DRB1 may inhibit the biological function of endothelial cells by targeting miRNA_12118 and acting on FLT − 1. by targeting miRNA_12118 and acting on FLT-1. Therefore, our study highlights the key role of the hsa-HLA-DRB1/miRNA_12118/FLT-1 axis in diabetic foot trauma.

PH and Sect. 7 domain-containing protein 3 (PSD3) has been reported to be associated with some cancers, but its role in esophageal squamous cell carcinoma (ESCC) has not been thoroughly examined. The purpose of this study was to investigate the expression of PSD3 in ESCC and determine whether PSD3 is regulated by pyruvate kinase type M2 (PKM2) to affect the malignant phenotype of ESCC cells. First, we found that PSD3 was highly expressed in ESCC tissues and correlated with ESCC lymph node metastasis. In vitro, PSD3 promoted the proliferation, migration and invasion of ESCC cells. In vivo, PSD3 accelerated ESCC growth and metastasis. Next, the interaction between PSD3 and PKM2 was examined, and the results showed that PSD3 was directly regulated by PKM2. Functionally, PSD3 was regulated by PKM2 to promote the proliferation, migration and invasion of ESCC cells. Mechanistically, PSD3 was regulated by PKM2 to upregulate the expression of Vimentin and Snail and downregulate the expression of E-cadherin. Collectively, all the data we show here demonstrate that PSD3, regulated by PKM2, endows growth and metastasis advantages in ESCC by modulating epithelial-mesenchymal transition (EMT) progression.

Background The activation imbalance of M1/M2 macrophage phenotypes is crucial in diabetic nephropathy (DN). This study aimed to explore the molecular mechanisms underlying quercetin's action against DN. Methods In vitro, RAW 264.7 macrophages were incubated with high glucose (HG) with or without quercetin. Overexpression of NLRC5 was investigated to elucidate the mechanism. M1/M2 macrophage differentiation was assessed by flow cytometry using cell surface markers CD86 and CD206. In vivo, a DN mouse model was created using a high-fat diet and streptozotocin (STZ). Quercetin was administered intragastrically to DN mice at 50 mg/kg and 100 mg/kg. After euthanasia, mouse kidneys were analyzed by hematoxylin and eosin (H&E), Masson's trichrome, and immunohistochemistry (IHC) staining. ELISA assay and western blot analysis were performed to determine related molecular levels. Results In vitro, quercetin significantly reduced HG-induced expressions of CD86, iNOS, NLRC5, NLRP3, and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β), while increasing HG-induced CD206, Arg-1, and IL-10 in RAW 264.7 macrophages. However, these effects of quercetin were abolished when NLRC5 was overexpressed. In DN mice, quercetin administration ameliorated renal histopathological injury and fibrosis. Notably, there was a significant reduction in expressions of NLRC5, NLRP3, Col1a1, and α-SMA, along with decreased expressions of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β). Conclusion This study showed that quercetin improves DN by inhibiting M1-type macrophages through targeting the NLRC5/NLRP3 pathway.

Objective:This study aimed to investigate whether exposure to multiple ambient air pollutants (PM1, PM2.5, PM10, O₃, and NO₂) elevates the risk of spinal osteoarthritis among middle-aged and older adults in China, and to further determine if there are gender-specific differences in vulnerability. Methods:A total of 7,663 participants aged 45 years and older, drawn from the China Health and Retirement Longitudinal Study (CHARLS), were followed from 2011 to 2020. Individuals free of spinal osteoarthritis at baseline were included. Annual mean concentrations of PM1, PM2.5, PM10, O₃, and NO₂ were extracted from the China High Air Pollutants (CHAP) dataset at a 1 km resolution (10 km for NO₂ in some years). Spinal osteoarthritis was identified via self-reported, physician-diagnosed cases involving the spine. Time-varying Cox proportional hazards regression models were used to evaluate hazard ratios (HR) and 95% confidence intervals (CI) per 10 μg/m3 increase in pollutant concentrations. All analyses accounted for demographic, socioeconomic, lifestyle, and spatial/seasonal factors, and explored potential effect modification by gender. Results:During the median 7-year (IQR: 4-9 years) follow-up, 1,556 participants newly reported spinal osteoarthritis. After adjusting for confounders, each 10 μg/m3 increment of PM1, PM2.5, PM10, and NO₂ was associated with a significant rise in the incidence of spinal osteoarthritis (13.8, 6.8, 5.1, and 17.4%, respectively), while O₃ showed a weaker and non-significant effect (1.1%). Notably, stratified analyses revealed that female participants exhibited pronounced vulnerability to PM1, PM2.5, PM10, and NO₂ exposures, whereas the associations in males were not statistically significant. Conclusion:This prospective study indicates that higher concentrations of particulate matter and traffic-related pollutants may contribute to an elevated risk of spinal osteoarthritis, particularly among women. These findings underscore the importance of incorporating musculoskeletal health into air quality management and highlight the value of targeted interventions-such as reducing ambient pollution and monitoring high-risk groups-to mitigate the burden of spinal osteoarthritis in rapidly urbanizing areas.

Objective: Acteoside (ACT) has different pharmacological properties such as antioxidant, hepatoprotective and anti-inflammatory effects. Impaired mitophagy has been recognized as an important pathogenic factor in metabolic dysfunction-associated steatotic liver disease (MASLD). Nevertheless, the possible therapeutic role of ACT in MASLD and the exact effect of ACT on mitophagy regulation are not explored. This study aims to elucidate the therapeutic efficacy of ACT in a high-fat and high-sugar (HFHS) diet-induced mouse model of MASLD and to determine whether its effects are related to the activation of PINK1/Parkin-related mitophagy markers. Methods: C57BL/6J mice were randomly allocated to control, model, rosuvastatin (RSF, 3 mg/kg), and ACT (30, 60, and 120 mg/kg) groups. Following a 14-week continuous intervention, biochemical parameters, liver histology, and mitophagy-related markers were assessed. Results: ACT administration significantly improved serum lipid profiles, liver function and insulin resistance, marked by reduced levels of MDA, IL-6, TNF-α, IL-1β, LDL-C, TC, TG, AST, ALT, HOMA-IR (p < 0.05), while increasing HDL-C and enhancing hepatic GSH-Px and SOD activities (p < 0.05). Histological examination revealed a notable attenuation of hepatic steatosis and lipid accumulation. At the molecular level, ACT promoted mitophagy activation, as indicated by upregulated PINK1, LC3II/I, and Parkin expression and downregulated P62 and p-P62. Electron microscopy further validated the restoration of mitochondrial morphology and reduction in lipid droplets. Conclusions: These results demonstrate that ACT ameliorates MASLD progression by improving metabolic homeostasis, reducing inflammation and oxidative stress, and alleviating PINK1/Parkin-related mitophagy impairment to restore mitophagy homeostasis. Our study highlights the potential of ACT as a new therapeutic agent for MASLD.

ObjectiveEndothelial dysfunction is a central contributor to the vascular complications observed in individuals with diabetes. cAMP response element-binding protein (CREB) plays a crucial role in mediating hyperglycemia-induced endothelial dysfunction. Phosphatase and tensin homolog (PTEN) has been implicated in the regulation of endothelial inflammation, yet the precise mechanism by which CREB modulates PTEN to protect endothelial cells under high glucose conditions remains unknown. This study aims to elucidate this potential mechanism.MethodsHuman umbilical vein endothelial cells (HUVECs) were exposed to high glucose (30 mM) or normal glucose (5.5 mM) for 6 days. Cell viability and apoptosis were assessed via the Cell Counting Kit-8 and flow cytometry. To evaluate oxidative stress, the levels of reactive oxygen species (ROS), lactate dehydrogenase (LDH), and malondialdehyde (MDA) were measured via commercial assay kits. The interaction between CREB and endothelial specific molecule 1 (ESE-1) was assessed via coimmunoprecipitation. Chromatin immunoprecipitation and luciferase reporter assays were used to investigate the transcriptional regulation of PTEN by ESE-1 and CREB. Western blotting was performed to analyze the expression of intercellular adhesion molecule-1 and E-selectin. The adhesion of HUVECs was evaluated via monocyte-endothelial cell adhesion assays.ResultsOur findings revealed a direct interaction between CREB and ESE-1, which together regulate PTEN expression to activate the phosphoinositide 3-kinase/protein kinase B pathway. Under high-glucose conditions, we observed significant increases in oxidative stress, inflammatory responses, and adhesion in HUVECs. ESE-1 knockdown reversed these effects, restoring endothelial cell function. Moreover, the overexpression of PTEN in high glucose-treated HUVECs rescued the endothelial injury induced by ESE-1 knockdown, suggesting that PTEN plays a pivotal role in mediating the protective effects.ConclusionESE-1, through the regulation of CREB-mediated PTEN expression, activates the PI3K/AKT pathway and modulates key processes such as oxidative stress, inflammation, and adhesion in endothelial cells under high-glucose stress.

Ethnopharmacological relevance: "Snow-white waterlily" (Nymphaea candida) dried flower possesses various efficacy in Uighur medicine such as reducing fever and nourishing the liver, anti-inflammatory and cough relieving, moistening the throat and quenching thirst. Aim of the study: Polyphenols are characteristic component of N. candida as well as its quality markers, and the purpose of this study was to conduct investigations into anti-inflammatory, antitussive, antipyretic, and analgesic activities of the polyphenol-enriched fraction from N. candida (NCTP) in order to validate the traditional efficacy of this plant. Materials and methods: The polyphenols in NCTP were analyzed by HPLC, and an acute oral toxicity study was conducted for NCTP. The anti-inflammatory activities of NCTP were evaluated using xylene induced ear edema, capillary permeability, cotton pellet granuloma, and carrageenan-induced rat paw edema, of which multiple biochemical indices were measured in carrageenan-induced rat paw edema such as prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2),5-lipoxygenase (5-LOX), interleukin-6 (IL-6), interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) activities; the analgesic activities were investigated using acetic acid writhing, hot plate test, and formalin test; the anti-tussive and antipyretic effects were tested by ammonia induced cough in mice and yeast-induced fever respectively. Results: NCTP with LD50 of 5222 mg/kg was low toxicity and safety. NCTP (200 mg/kg) could significantly reduce ear swelling and capillary permeability by 30.63% and 31.37%, respectively. NCTP revealed 15.76% inhibiting activities in cotton pellet granuloma in mice at a dosage of 200 mg/kg. Furthermore, NCTP (50, 100, and 200 mg/kg) substantially decreased carrageenin-induced paw edema in rats between 1 and 5 h, and NCTP could decrease PGE2, 5-LOX, COX-2 levels as well as IL-6, IL-1 beta, TNF-alpha activities compared with the control group; NCTP could decrease MDA contents in carrageenin-induced rise, and increase SOD and GSH activities. Furthermore, the dose-dependent inhibition effect of NCTP on pain was revealed in the hot plate experiment. In addition to reducing the amount of writhes brought on by acetic acid, NCTP (50, 100, and 200 mg/kg) significantly inhibited pain latency against both stages of the formalin test. Moreover, NCTP (50, 100, 200 mg/kg) showed the better antitussive activities in mice in a dose-dependent manner. In the yeast-induced pyrexia test, dosages of 50, 100, and 200 mg/kg resulted in a statistically significant drop in rectal temperature. Conclusion: The experimental results proved the analgesic, anti-inflammatory, anti-tussive and antipyretic activities of the polyphenol-enriched fraction from N. candida, and supported the traditional use of this plant as well.

Recent studies have increasingly focused on PIK3CA mutations in esophageal squamous cell carcinoma (ESCC); however, the clinicopathological significance of these mutations within the tumor microenvironment remains underexplored. This study aimed to evaluate and compare the clinicopathological significance of mutated PIK3CA in ESCC using in silico analyses of the ESCC dataset from the TCGA database. We assessed prognosis, differential expression, correlation with immune cell infiltration and immune checkpoint expression, heterogeneity, and drug sensitivity in comparison with wild-type PIK3CA. Our findings revealed that PIK3CA mutation is associated with increased tumor mutation burden and significantly correlated with the infiltration of CD4 naive and effector memory CD4 T cells. Additionally, ESCC cells harboring PIK3CA mutations exhibited reduced sensitivity to p38/JNK MAPK inhibitors compared to those with wild-type PIK3CA. Collectively, our in silico analysis suggests that mutational PIK3CA plays a role in resistance to p38 and JNK MAPK inhibitors in ESCC.

This study aimed to investigate the effect of phenylethanol glycoside from Cistanche tubulosa (CPhGs) on the prevention of bovine serum albumin (BSA)-induced hepatic fibrosis in rats. Investigation of the mechanisms of the anti-hepatic fibrosis effect was focused on CPhGs’ influence on the “gut–liver” regulation, including the gut microbiota, intestinal barrier, systemic lipopolysaccharide (LPS) concentration, and LPS-related signaling pathway. The results show that CPhGs restored the diversity of gut microbiota, increased the relative abundance of Bacteroidetes, and decreased the relative abundance of Firmicutes and Proteobacteria in the fibrotic rats. In addition, CPhGs promoted the enrichment of probiotics such as Blautia, Oscillospira, Ruminococcus, Odoribacter, Bacteroides, and Parabacteroides in intestines of these rats. Furthermore, CPhGs reduced histopathological injury in the intestine and restored the tight junctions of the intestine by increasing the expression of ZO-1, occludin, and E-cadherin. CPhGs efficiently reduced serum LPS and liver lipopolysaccharide-binding protein (LBP) levels and inhibited the LPS-TLR4/MyD88/NF-κB pathway, which is related to protein expression in the liver. Correlation analysis confirmed that these beneficial bacteria were negatively associated with pathological damage, while LPS and harmful bacteria were positively associated with liver injury. Our fecal microbiota transplantation (FMT) experiment confirmed that gut microbiota is an important part of disease progression and that CPhGs is useful for the prevention and treatment of hepatic fibrosis. Our data demonstrate that the anti-hepatic fibrosis mechanism of CPhGs was mediated by regulation of the “gut–liver” axis. These results can stimulate consideration for its use in clinical practices.

Short for pyruvate kinase M2 subtype, PKM2 can be said of all‐round player that is notoriously known for its metabolic involvement in glycolysis. Holding a dural role as a metabolic or non‐metabolic (kinase) enzyme, PKM2 has drawn extensive attention over its biological roles implicated in tumor cells, including proliferation, migration, invasion, metabolism, and so on. wandering PKM2 can be transboundary both intracellularly and extracellularly. Specifically, PKM2 can be nuclear, cytoplasmic, mitochondrial, exosomal, or even circulate within the body. Importantly, PKM2 can function as an RNA‐binding protein (RBP) to self‐support its metabolic function. Despite extensive investigations or reviews available surrounding the biological roles of PKM2 from different angles in tumor cells, little has been described regarding some novel role of PKM2 that has been recently found, including, for example, acting as RNA‐binding protein, protection of Golgi apparatus, and remodeling of microenvironment, and so forth. Given these findings, in this review, we summarize the recent advancements made in PKM2 research, mainly from non‐metabolic respects. By the way, PKM1, another paralog of PKM2, seems to have been overlooked or under‐investigated since its discovery. Some recent discoveries made about PKM1 are also preliminarily mentioned and discussed.