Sepsis-associated encephalopathy (SAE) is a diffuse cerebral dysfunction caused by sepsis-triggered systemic immune disorders. It has a high incidence and lacks specific diagnostic and therapeutic strategies. This study aimed to investigate whether cytidine/uridine monophosphate kinase 2 (CMPK2) contributes to SAE pathogenesis by regulating neuroinflammation. Bioinformatic analysis identified CMPK2 as a core hub gene for SAE progression, with its expression significantly upregulated in SAE mouse brain tissues and lipopolysaccharide-activated microglia. In vivo experiments showed that the CMPK2 inhibitor nordihydroguaiaretic acid (NDGA) significantly alleviated neuroinflammation and improved cognitive impairments in SAE mice, revealing NDGA’s potential therapeutic effect on SAE. In vitro, NDGA intervention or viral-mediated CMPK2 silencing markedly reduced oxidative stress and DNA damage in microglia, ameliorated mitochondrial dysfunction, and inhibited abnormal activation of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway, thereby synergistically relieving proinflammatory activation of microglia. Rescue assays revealed that silencing CMPK2 reversed the excessive inflammation and cGAS-STING hyperactivation induced by CMPK2 overexpression in microglia, confirming that CMPK2 mediates neuroinflammation via the cGAS-STING axis. Mechanistically, ATF2 was predicted as an upstream transcription factor of CMPK2. CUT&Tag and dual-luciferase assays validated that ATF2 binds to the CMPK2 promoter to enhance its transcription, thereby activating the cGAS-STING pathway and aggravating neuroinflammation. Collectively, these findings identify the ATF2-CMPK2-cGAS-STING signaling cascade as a pivotal regulatory mechanism governing microglial inflammatory activation in SAE progression, providing a novel molecular mechanism and promising therapeutic target for SAE treatment.

Efferocytosis plays a vital role in mitigating inflammation and subsequent tissue damage by clearing dying cells after intracerebral hemorrhage, but its underlying mechanism remains poorly understood. In this study, programmed cell death 4 (PDCD4) exhibited upregulated expression and localization to microglia following intracerebral hemorrhage. Targeted adenoviral-mediated Pdcd4 knockdown in vivo markedly improved neurological recovery and enhanced microglial efferocytosis in intracerebral hemorrhage model mice. These effects were further validated in Pdcd4 knockout mice. In an in vitro intracerebral hemorrhage model using oxyhemoglobin-treated microglia, Pdcd4 knockdown notably increased the expression of efferocytosis-related molecules AXL receptor tyrosine kinase (AXL) and MER proto-oncogene, tyrosine kinase (MERTK). Mechanistically, an interaction between PDCD4 and signal transducer and activator of transcription 3 (STAT3) identified under basal conditions was amplified following intracerebral hemorrhage. Pdcd4 knockdown also promoted Janus kinase 2 (JAK2)-mediated STAT3 phosphorylation and nuclear translocation, driving Mertk and Axl transcription to facilitate microglial efferocytosis. Furthermore, Pdcd4 knockdown markedly reduced proinflammatory factor expression while enhancing anti-inflammatory factor levels, thereby mitigating the neuronal apoptosis induced by conditioned medium. These findings indicate that PDCD4 blockade enhances microglial efferocytosis by activating the JAK2-STAT3-Mertk/Axl pathway and promotes post-intracerebral hemorrhage neurological recovery, identifying PDCD4 as a potential therapeutic target.

Quercetin, a kind of flavonoids, has been implicated in multiple neurological diseases. Nevertheless, the functional roles and mechanisms of quercetin in HIV-associated neurocognitive disorders (HAND) remain unclear. HIV-1-encoded transactivator of transcription (Tat) is the major pathogenic factor for the progression of HAND. In the central nervous system, microglia-mediated Tat neurotoxicity is mainly comprised of inflammatory response, nitric oxide (NO) and excessive glutamate. In this study, we demonstrated that Tat-activated NF-κB p65 directly induce the release of IL-6 and TNF-α as well as NO production, while Tat promoted glutamate release via NF-κB/SPI1 pathway. Conversely, quercetin could upregulate SIRT3 expression to reduce reactive oxygen species (ROS) generation, thereby inhibiting NF-κB/SPI1 pathway and mitigating microglia-mediated Tat neurotoxicity. In addition, we also observed that quercetin alleviated neuronal apoptosis induced by the microglia-derived conditioned media in a SIRT3/ROS/NF-κB-dependent manner. Furthermore, Tat was found to downregulate SIRT3 expression via NF-κB/SPI1 pathway, which was reversed by quercetin in microglia. Thus, our data establish that quercetin disrupts the SIRT3/ROS/NF-κB/SPI1 feedback loop to attenuate microglia-mediated Tat neurotoxicity. Finally, in murine models, we recapitulated that administration of quercetin remarkably ameliorated Tat‑induced neuropathy and cognitive decline in vivo. Taken together, our study uncovers the neuroprotective roles of quercetin in the amelioration of microglia-mediated Tat neurotoxicity and highlights its potential as a therapeutic agent for the treatment of HAND.

AIM: To investigate the role of pyruvate kinase M2 (PKM2) in high glucose (HG)-stimulated retinal endothelial cells and its underlying molecular mechanisms and signaling pathways in retinal angiogenesis. METHODS: Human retinal microvascular endothelial cells (HRMECs) were cultured and divided into the following groups: normal glucose (NG, 5.5 mmol/L), HG (30 mmol/L), HG with PKM2 knockdown (HG+shPKM2), and HG treated with the pharmacological activator TEPP-46 (HG+TEPP-46). Cellular viability, proliferation, migration, and tube-forming ability were assessed using CCK-8, EdU, wound healing/Transwell, and Matrigel assays, respectively. The expression levels of PKM2, phosphorylated PKM2 (p-PKM2, Y105), hypoxia-inducible factor-1α (HIF-1α), and vascular endothelial growth factor A (VEGFA) were detected by Western blotting. The oligomerization status of PKM2 was analyzed via native gel electrophoresis. The subcellular localization of PKM2 was examined by immunofluorescence and nuclear-cytoplasmic fractionation. RESULTS: Under HG stimulation, the expression level of PKM2 was significantly increased (P<0.05). Knockdown of PKM2 was found to markedly suppress cell viability, proliferation, migration, and tube formation in HRMECs (P<0.05). Mechanistic studies revealed that phosphorylation of PKM2 at the Y105 site was promoted by HG treatment, which induced its dissociation from a tetramer to a dimer, thereby driving its nuclear translocation. Upon entering the nucleus, PKM2 was shown to exert critical non-metabolic functions; it was physically bound to HIF-1α and acted as its co-activator, leading to significant upregulation of VEGFA expression (P<0.05). In contrast, the PKM2 activator TEPP-46 effectively prevented dimerization and nuclear translocation of PKM2 by promoting its tetramerization. Consequently, the PKM2/HIF-1α axis-mediated upregulation of VEGFA was blocked, ultimately resulting in the reversal of HG-induced angiogenesis. CONCLUSION: HG influences retinal endothelial cell function by inducing PKM2 phosphorylation, dimerization, and nuclear translocation. The shift in PKM2 phosphorylation and oligomerization status represents a key mechanism through which TEPP-46 reverses HG-induced angiogenesis.

PI3K complex consists of catalytic subunit p110s and regulatory subunit p85s. Emerging evidence indicates that p110-free p85 subunits play pivotal roles in diverse biological processes, including cancer progression. In this study, we demonstrate the underlying mechanism of p110-free p85β in hepatocellular carcinoma (HCC) development. PIK3R2/p85β is upregulated in HCC and correlates with poor patient survival. Nuclear p85β, but not its cytoplasmic counterpart, exhibits oncogenic activity. In the nucleus of HCC cells, p85β undergoes liquid-liquid phase separation (LLPS) and specifically accumulates in the fibrillar centers of nucleoli, where it drives HCC progression. Within the nucleolar compartment, p85β interacts with and stabilizes POLR1A, the catalytic core subunit of RNA polymerase I, thereby enhancing rRNA biosynthesis and maintaining HCC stemness. Furthermore, we develop an engineered circular RNA that encodes a peptide containing p110α ABD domain, which effectively suppresses HCC tumor growth by simultaneously disrupting p85β/POLR1A condensates and inhibiting PI3K/AKT signaling pathway, offering a novel RNA-based therapeutic strategy against HCC.

Heat shock factor 2 (HSF2) and hypoxia-inducible factor 1α are activated by angiotensin II (ANGII) in cardiomyocytes. The endoplasmic reticulum (ER) stress plays a critical role in cardiac hypertrophy. Moreover, HIF-1α is known to be regulated by HSF2 in tumour cells. In this study, we hypothesised and clarified whether HSF2 trans-activated HIF-1α through initiation of ER stress in hypertrophic cardiomyocytes. Myocardial hypertrophy was induced by the treatment of ANGII. Expression of the gene or protein was assessed by applying RT-PCR, WB, ICC and IHC. Luciferase and CHIP were applied to detect the transcription of HIF-1α by HSF2. Both in vitro and in vivo, the expression of HIF-1α, ER stress markers and HSF2 was increased in ANGII-treated hypertrophic cardiomyocytes. Blocking ER stress suppressed the expression of HSF2 and HIF-1α in ANGII-treated cardiomyocytes. Silencing HSF2 inhibited HIF-1α, thereby reducing hypertrophy but had no effect on ER stress. Similarly, silencing HIF-1α reduced hypertrophy without affecting ER stress or HSF2 expression. HSF2 transcriptionally activated HIF-1α. We concluded that ER stress induced by ANGII activates HSF2, which then trans-activates HIF-1α, promoting cardiac hypertrophy.

Background Myasthenia gravis (MG) is an autoimmune neuromuscular disorder in which immune dysregulation and altered T-cell homeostasis contribute to pathogenic autoantibody production. Jianpi Yiqi Busui Formula (JPYQBSF) is used as an adjunctive traditional Chinese medicine, but its immunometabolic mechanisms remain incompletely defined. Purpose To evaluate the clinical and preclinical effects of JPYQBSF and to examine whether gut microbiota remodeling and the regulatory T-cell (Treg) LDHA-lactate-H3K18la-PD-1 axis are associated with its immunomodulatory activity. Study design An exploratory prospective clinical cohort, an experimental autoimmune myasthenia gravis (EAMG) rat model, and multi-omics and mechanistic experiments were integrated. Methods Thirty-one anti-acetylcholine receptor antibody-positive MG patients receiving stable conventional therapy were assessed before and after 24 weeks of adjunctive JPYQBSF, with 31 age- and sex-matched healthy participants as a reference group. EAMG rats underwent functional, electrophysiological, immunological, 16S rRNA, serum metabolomic, and TMT-proteomic analyses. Fecal microbiota transplantation, human Treg-cell experiments, pharmacological LDH inhibition, LDHA knockdown, ChIP-qPCR, CUT&Tag, and reporter assays were used to interrogate candidate pathways. Results In the clinical cohort, adjunctive JPYQBSF was associated with lower AChR-Ab levels and partial normalization of inflammatory and immunoregulatory markers; the uncontrolled design precludes attribution of these changes to JPYQBSF alone. In EAMG rats, JPYQBSF improved motor and electrophysiological outcomes, reduced inflammatory mediators, and shifted the Treg/Th17 balance. Microbiome, metabolomic, and proteomic analyses identified treatment-associated community and metabolic changes, including aromatic lactic-acid derivatives and LDHA. In cultured Tregs, JPYQBSF exposure increased LDHA expression, lactate, H3K18la enrichment near the PDCD1 promoter, and PD-1 expression. Oxamate and LDHA knockdown attenuated several of these changes, whereas healthy-donor FMT partially reproduced selected immunometabolic effects. Conclusion The findings support a coordinated model in which gut microbiota remodeling and LDHA-dependent Treg immunometabolic changes contribute to the effects of JPYQBSF. They do not establish a direct linear causal pathway from specific microbial metabolites to H3K18 lactylation, PD-1 regulation, or independent clinical efficacy.

Psoriasis is a chronic inflammatory skin disease characterized by excessive keratinocyte proliferation and dysregulated immune responses. The transcriptional regulator AFF3 (ALF transcription elongation factor 3) has recently been implicated in immune-related diseases; however, its role in psoriasis remains unclear. Single-cell sequencing was used to assess AFF3 expression in psoriatic keratinocytes, and immunohistochemical analysis was performed on skin tissue samples from five patients with psoriasis and five healthy controls. A psoriatic inflammation model was established by stimulating HaCaT cells and primary mouse keratinocytes with an M5 cytokine cocktail (IL-17 A, IL-22, IL-1α, TNF-α, and oncostatin M). AFF3 expression was quantified by RT-qPCR and Western blotting. AFF3 knockdown was achieved via siRNA-mediated lentiviral transduction, and functional effects were assessed using CCK-8 assay, flow cytometry, and quantification of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Downstream signaling pathways were identified by transcriptomic sequencing, and activation of the MAPK signaling pathway was evaluated by Western blotting. AFF3 expression was elevated in keratinocytes from psoriatic skin lesions compared with normal skin. In both HaCaT cells and primary mouse keratinocytes, AFF3 expression was significantly upregulated following LPS or M5 stimulation. AFF3 knockdown inhibited M5-induced keratinocyte proliferation, increased apoptosis, and reduced oxidative stress, pro-inflammatory cytokine release, and aberrant expression of KRT16, KRT17, and KRT19. Mechanistically, AFF3 knockdown led to decreased phosphorylation levels of ERK, p38, and JNK, indicating suppression of the MAPK signaling pathway. These findings indicate that AFF3 knockdown attenuates keratinocyte hyperproliferation and psoriasis-like inflammatory responses, at least in part, through inhibition of the MAPK signaling pathway.

AIM To investigate the role of pyruvate kinase M2 (PKM2) in high glucose (HG)-stimulated retinal endothelial cells and its underlying molecular mechanisms and signaling pathways in retinal angiogenesis. METHODS Human retinal microvascular endothelial cells (HRMECs) were cultured and divided into the following groups: normal glucose (NG, 5.5 mmol/L), HG (30 mmol/L), HG with PKM2 knockdown (HG+shPKM2), and HG treated with the pharmacological activator TEPPu201146 (HG+TEPPu201146). Cellular viability, proliferation, migration, and tubeu2011forming ability were assessed using CCKu20118, EdU, wound healing/Transwell, and Matrigel assays, respectively. The expression levels of PKM2, phosphorylated PKM2 (pu2011PKM2, Y105), hypoxia-inducible factor-1u03B1 (HIFu20111u03B1), and vascular endothelial growth factor A (VEGFA) were detected by Western blotting. The oligomerization status of PKM2 was analyzed via native gel electrophoresis. The subcellular localization of PKM2 was examined by immunofluorescence and nuclearu2011cytoplasmic fractionation. RESULTS Under HG stimulation, the expression level of PKM2 was significantly increased (Pu0026lt;0.05). Knockdown of PKM2 was found to markedly suppress cell viability, proliferation, migration, and tube formation in HRMECs (Pu0026lt;0.05). Mechanistic studies revealed that phosphorylation of PKM2 at the Y105 site was promoted by HG treatment, which induced its dissociation from a tetramer to a dimer, thereby driving its nuclear translocation. Upon entering the nucleus, PKM2 was shown to exert critical nonu2011metabolic functions; it was physically bound to HIFu20111u03B1 and acted as its cou2011activator, leading to significant upregulation of VEGFA expression (Pu0026lt;0.05). In contrast, the PKM2 activator TEPPu201146 effectively prevented dimerization and nuclear translocation of PKM2 by promoting its tetramerization. Consequently, the PKM2/HIFu20111u03B1 axisu2011mediated upregulation of VEGFA was blocked, ultimately resulting in the reversal of HGu2011induced angiogenesis. CONCLUSION HG influences retinal endothelial cell function by inducing PKM2 phosphorylation, dimerization, and nuclear translocation. The shift in PKM2 phosphorylation and oligomerization status represents a key mechanism through which TEPP-46 reverses HG-induced angiogenesis.

Background Multidrug resistance (MDR) poses a significant obstacle to developing chemotherapeutic treatments. In previous studies using a traditional model of adriamycin resistance (ADR) with K562 cells, we demonstrated that N-acetylglucosaminyltransferase III (GnT-III) expression negatively regulates chemoresistance. Additionally, we observed that fucosylation levels were increased in the ADR cells. Method Fucosylation levels were determined using lectin blot, western blot, and flow cytometry. Gene expression levels were analyzed via qPCR. We generated a FUT4 knockout (KO) ADR cell line using CRISPR/Cas9 technology. Cytotoxicity and drug efflux assays were conducted to evaluate chemotherapy tolerance. Results The expression levels of FUT4 and its products, the LeX antigens, were significantly upregulated in the ADR cells compared to the parental K562 cells. The FUT4 KO reduced the elevated levels of P-glycoprotein (P-gp) found in ADR cells and exhibited increased sensitivity to chemotherapeutic drugs. Furthermore, restoring FUT4 expression in the KO cells effectively reversed P-gp expression, drug efflux, and chemoresistance. Given the critical role of the NF-κB pathway in P-gp expression, we investigated NF-κB signaling and found that the phosphorylation levels of p65 were significantly increased in the ADR cells but were downregulated in the FUT4 KO cells. Furthermore, the restoration of FUT4 rescued the phosphorylation levels of p65. Conclusions FUT4 specifically upregulates P-gp expression related to chemoresistance through the NF-κB signaling pathway. General significance This study highlights the importance of FUT4 in chemoresistance and suggests it may serve as a promising target for combating MDR.

OBJECTIVE:The present study aims to explore the role of nafamostat mesylate (NM) in ischemia-reperfusion (I/R)-induced acute kidney injury (AKI). METHODS:Twenty-one male rats were randomly divided into three groups: sham, I/R, and I/R + NM groups. Acute renal I/R injury models were created in the I/R and I/R + NM groups by right nephrectomy and clamping the left renal pedicle for 45 min. Rats in the I/R + NM group were intraperitoneally injected with 0.75 mg/kg of NM before modeling. Blood and kidney specimens were collected at 24 h after model establishment. RNA sequencing (RNA-seq), KEGG and GO enrichment analyses, and single-cell sequencing were carried out to investigate the related mechanisms. RESULTS:The pretreatment with NM improved renal function, reduced mitochondrial damage, suppressed the accumulation of reactive oxygen species, and inhibited ferroptosis. The RNA-seq indicated that NM induced protection by upregulating fatty acids, inhibiting inflammation, and promoting DNA repair. Proximal tubular cells were mainly affected. CONCLUSION:NM protects against AKI by reducing reactive oxygen species, upregulating fatty acids, and regulating ferroptosis signaling pathways in proximal tubular cells.

Microglial pyroptosis and neuroinflammation have been implicated in the pathogenesis of sepsis-associated encephalopathy (SAE). OGT-mediated O-GlcNAcylation is involved in neurodevelopment and injury. However, its regulatory function in microglial pyroptosis and involvement in SAE remains unclear. In this study, we demonstrated that OGT deficiency augmented microglial pyroptosis and exacerbated secondary neuronal injury. Furthermore, OGT inhibition impaired cognitive function in healthy mice and accelerated the progression in SAE mice. Mechanistically, OGT-mediated O-GlcNAcylation of ATF2 at Ser44 inhibited its phosphorylation and nuclear translocation, thereby amplifying NLRP3 inflammasome activation and promoting inflammatory cytokine production in microglia in response to LPS/Nigericin stimulation. In conclusion, this study uncovers the critical role of OGT-mediated O-GlcNAcylation in modulating microglial activity through the regulation of ATF2 and thus protects against SAE progression.

Neuropathic pain is a chronic condition characterized by damage to and dysfunction of the peripheral or central nervous system. There are currently no effective treatment options available for neuropathic pain, and existing drugs often provide only temporary relief with potential side effects. Multilineage-differentiating stress-enduring (Muse) cells are characterized by high expansion potential, a stable phenotype and strong immunosuppression. These properties make them attractive candidates for therapeutics for neuropathic pain management. In this study, we conducted a series of experiments to evaluate the effect of Muse cells on neuropathic pain. Muse cells from different species demonstrated analgesic potential by reversing CCI-induced neuropathic pain. Protein profiling revealed a high degree of similarity between Muse cells and BMSCs. The intrathecal injection of Muse cells effectively reduced neuropathic pain in various mouse models, resulting in better analgesic effects than the administration of equivalent low doses of BMSCs. Immunohistochemical analysis and qPCR revealed the ability of Muse cells to inhibit spinal cord neuroinflammation caused by SNI. In addition, Transwell and ELISA revealed that Muse cells migrated through the injured dorsal root ganglion (DRG) via the CCR7-CCL21 chemotactic axis. In addition, the secretion of TGF-b and IL-10 by Muse cells was identified as the mechanism underlying the analgesic effect of Muse cells. The capacity of Muse cells to mitigate neuroinflammation and produce analgesic effects via the modulation of TGF-b and IL-10 underscores their potential as promising therapeutic approaches for the treatment of neuropathic pain.

Intracerebral hemorrhage (ICH), a common neurological disorder with a high rate of disability, involves complex immunoinflammatory mechanisms, particularly those related to secondary inflammatory injury. Neutrophils, as the earliest subtype of leukocytes recruited after stroke, play a pivotal role in secondary brain injury. Traditionally, neutrophils were thought to mediate tissue damage primarily via phagocytosis, chemotaxis, and degranulation. However, recent studies have shown that neutrophils also contribute to the pathogenesis of intracerebral hemorrhage by releasing neutrophil extracellular traps (NETs), which exacerbate blood-brain barrier disruption, amplify local inflammy -30ation, and promote neuronal injury. This review systematically examines the interactions between the central and peripheral immune systems following ICH. It focuses on the bidirectional regulatory relationship between microglia and neutrophils, and their coordinated roles in inflammation, blood-brain barrier disruption, neurological dysfunction, and cognitive impairment. In addition, this review summarizes recent potential therapeutic strategies targeting the formation and clearance of NETs, including peptidylarginine deiminase 4 inhibitors, reactive oxygen species inhibitors, histone inhibitors, and DNases. These interventions may offer theoretical insights into novel therapeutic targets for mitigating secondary injury following ICH.

To determine the impact of sodium butyrate on the activation of the reactive oxygen species (ROS)/nuclear factor kappa B (NF-κB)/NLR family pyrin domain containing 3 (NLRP3) signaling pathway and angiogenesis in human retinal microvascular endothelial cells (HRMECs) caused by high glucose (HG). HRMECs were grown for 24 h or 72 h in HG solution (30 mmol/L D-glucose) with 5 mM NaB. Using Cell Counting Kit-8, the effects of HG and NaB levels on the viability of HRMECs were examined. Using various kits, intracellular ROS levels, lactate dehydrogenase (LDH), and Malondialdehyde (MDA) in cell supernatants were measured. Western blot, Immunofluorescence, and Real-time quantitative polymerase chain reaction were employed to quantify protein and messenger RNA expression. Using wound-healing and tube formation tests, the migratory proficiency and angiogenesis of HRMECs were evaluated. NaB demonstrated a reduction in ROS production, as well as the release of LDH and MDA in HG-induced HRMECs. Additionally, NaB led to a decrease in protein expression of phosphorylation of NF-κB, NLRP3, Caspase 1, interleukin, vascular cell adhesion molecule-1 and intercellular adhesion molecule-1. The impact of HG on zonula occluden-1, a tight junction protein, was attenuated by NaB. Furthermore, NaB inhibited the migration and tube formation of HRMECs partly by ROS/NF-κB/NLRP3 pathway. NaB suppresses the activation of ROS/NF-κB/NLRP3 signaling pathway and angiogenesis in HRMECs induced by HG.

Background: The hypothalamic feeding circuit is highly vulnerable to obesity-inducing diets, as observed in diet-induced obesity (DIO) models. Programmed cell death factor 4 (PDCD4) is widely expressed in various tissues and organs. Depending on the context, it exhibits both pro-inflammatory and anti-inflammatory properties. This study aimed to analyze serum PDCD4 levels and investigate its correlation with hypothalamic inflammation in obesity. Methods: A total of 195 participants were separated into two groups according to their body mass index (BMI): normal weight group (18.5 kg/m(2) <= BMI < 24 kg/m(2)) and obesity group (BMI >= 28 kg/m(2)). Serum levels of the following were measured using enzyme-linked immunosorbent assay (ELISA): PDCD4, neuropeptide Y (NPY), Ionized calcium-binding adapter molecule 1 (Iba1), and NOD-like receptor thermal protein domain-associated protein 3 (NLRP3). Other biochemical indicators were analyzed. Statistical analyses were performed to evaluate the association between serum PDCD4 levels and other biochemical indicators. Results: A significant increase in serum PDCD4 level was evident in the obesity group compared with the control group. A binary logistic regression analysis revealed a statistically significant relationship between PDCD4 and obesity (p < 0.05). Based on Spearman correlation analysis, a positive correlation was found between the serum PDCD4 level and BMI, and the serum PDCD4 level was positively correlated with NPY, Iba1 and NLRP3 levels (p < 0.05). Furthermore, serum PDCD4 level was found to be independently associated with Iba1 and NLRP3, indicating the role of PDCD4 in regulating hypothalamic inflammation in the obesity context. Conclusion: In addition to a significant elevation in obese patients, serum PDCD4 level is independently correlated with Iba1 and NLRP3 levels, suggesting a mediating role of PDCD4 in the activation of Iba1 and NLRP3, which is crucial for facilitating peripheral-to-central inflammatory crosstalk and ultimately the occurrence of hypothalamic inflammation.

Cutaneous squamous cell carcinoma (cSCC) is a malignant tumor originating from epidermal or appendageal keratinocytes, with a rising incidence in recent years. Understanding the molecular mechanism driving its development is crucial. This study aims to investigate whether miR-34a-5p is involved in the pathogenesis of cSCC by targeting Sirtuin 6 (SIRT6).The expression levels of miR-34a-5p and SIRT6 were determined in 15 cSCC tissue specimens, 15 normal tissue specimens and cultured cells via real-time polymerase chain reaction (RT-qPCR). Pearson's correlation analysis was conducted to evaluate the relationship between miR-34a-5p and SIRT6 expression levels in cSCC tissues. A431 and SCL-1 cells were transfected with miR-34a-5p mimic, negative control or miR-34a-5p mimic together with recombinant plasmids containing SIRT6 gene. Cell counting kit-8, clone formation assay, wound healing assay, and flow cytometry were employed to assess the effects of these transfections on proliferation, migration, and apoptosis, respectively. The interaction between miR-34a-5p and SIRT6 was characterized using a dual-luciferase reporter assay.MiR-34a-5p expression was down-regulated in cSCC tissues significantly, while the SIRT6 expression was the opposite. A negative correlation was observed between the expression of miR-34a-5p and SIRT6 in cSCC tissues. Furthermore, overexpression of miR-34a-5p led to a significant reduction in the proliferation and migration abilities of A431 and SCL-1 cells, accompanied by an increase in apoptosis levels and a decrease in SIRT6 expression levels. MiR-34a-5p was identified as a direct target of SIRT6. Importantly, overexpression of SIRT6 effectively counteracted the inhibitory effect mediated by miR-34a-5p in cSCC cells.Our findings suggest that miR-34a-5p functions as a tumor suppressor in cSCC cells by targeting SIRT6.

Alterations in anion balance potential, along with the involvement of cation-chloride cotransporters, play pivotal roles in the development of hyperalgesia after peripheral nerve injury. Chloride voltage-gated channel seven (CLCN7) is the predominant member of the CLC protein family. Investigations on CLCN7 have focused primarily on its involvement in osteosclerosis and lysosomal storage disorders; nevertheless, its contribution to neuropathic pain has not been determined. In this investigation, we noted high expression of CLCN7 in neurons situated within the spinal dorsal horns and dorsal root ganglions (DRGs). Immunofluorescence analysis revealed that CLCN7 was predominantly distributed among IB4-positive and CGRP-positive neurons. Furthermore, the expression of CLCN7 was observed to be mainly reduced in neurons within the spinal dorsal horns and in small- and medium-sized neurons located in the DRGs of spared nerve injury mice. Knockdown of CLCN7 via siRNA in the DRGs resulted in increased mechanical and thermal hyperalgesia in naïve mice. Furthermore, the excitability of cultured DRG neurons in vitro was augmented upon treatment with CLCN7 siRNA. These findings suggested that CLCN7 downregulation following SNI was crucial for the manifestation of mechanical and thermal hyperalgesia, highlighting potential targeting strategies for treating neuropathic pain.

IntroductionDiabetic macular edema (DME) is a major cause of vision loss in the sick with diabetic retinopathy. The occurrence of DME is closely related to the breakdown of neurovascular coupling; however, its underlying mechanism has not been fully elucidated. The aim of this study was to investigate the diagnostic biomarkers and potential molecular mechanisms associated with neurovascular coupling in DME.MethodsThe differential expression analysis, STEM, and WGCNA were performed from GSE160306 to identify hub genes. The gene expression was validated by RT-qPCR. The relevant mechanisms of action were investigated through GO, KEGG, and GSEA analyses, as well as co-expression networks. Additionally, the LASSO regression analysis and a nomogram were used to demonstrate the diagnostic effectiveness of the model. Finally, the GenDoma platform was utilized to identify drugs with potential therapeutic effects on DME.ResultsNeurotrophic factor receptor (NGFR) was identified as a hub gene related to neurovascular coupling and DME. The expression of NGFR was verified by RT-qPCR in vitro cells. GSEA analysis indicated that high expression of NGFR may affect immunity and inflammatory pathway, thereby regulating neurovascular coupling and mediating the development of DME. The NGFR co-expression network was constructed, which exhibited the correlation with the neurotrophin signaling pathway. Moreover, a diagnostic model for DME based on NGFR and PREX1 demonstrated relatively good diagnostic performance using LASSO regression analysis and the nomogram. And then the GenDoma platform identified drugs with potential therapeutic effects on DME.ConclusionThe high expression of NGFR may lead to abnormal neurovascular coupling and participate in the occurrence of DME by regulating the immunity, inflammatory and neurotrophin signaling pathway. Detection of NGFR and related expression genes may be beneficial for monitoring the occurrence and development of DME.

BackgroundCutaneous squamous cell carcinoma (CSCC) is one of the most common types of skin cancer worldwide. Therefore, the identification of biomarkers associated with CSCC progression could aid in the early detection of high-risk squamous cell carcinoma and the development of novel therapeutic strategies.ObjectiveThis study aimed to investigate the expression patterns of silent mating type Information Regulation 2 homolog 6 (SIRT6) in CSCC and its clinical significance.MethodsThe protein expression level of SIRT6 in tissues was detected by immunohistochemistry, and the correlation between SIRT6 expression and clinicopathological parameters in CSCC patients was analyzed. The relative expression of SIRT6 in CSCC cell lineage and tissue specimens was determined by western blotting and PCR. The effect of SIRT6 silencing on cell proliferation was evaluated using cell counting kit 8. Wound healing, transwell method, and flow cytometry were used to investigate the migration, invasion, and cell cycle distribution/apoptosis of CSCC cells after SIRT6 silencing, respectively. Western blot was used to detect the expression of EMT (Epithelial-Mesenchymal Transition), cycle, apoptosis, and other related proteins.ResultsThe high expression of SIRT6 was correlated with the location of cancer tissue and Broder staging in CSCC patients. Knockdown of SIRT6 inhibited the proliferation, migration, invasion and EMT of CSCC cells, and promoted their apoptosis, with cells blocked in G1 phase.Study limitationsNo animal experiments were conducted to further verify the results.ConclusionDecreased expression of SIRT6 can inhibit the occurrence and development of CSCC.