The spread of MDR-GNB producing both SBLs and MBLs has created an urgent unmet clinical need, as current therapies fail to address both enzyme-mediated resistance and infection-induced inflammation. Ebselen is a selenium-containing heterocyclic compound and has been identified as a novel covalent inhibitor of MBLs. Guided by structural optimization and SAR analysis, we introduced a boronic acid group as a covalent warhead targeting SBLs and finally obtained E3f. E3f exhibited potent inhibitory activity against NDM-1 (MBLs, IC50 = 1.8 μM) and KPC-2 (SBLs, IC50 = 2.6 μM) by forming covalent adducts with Cys221 of NDM-1 and Ser70 of KPC-2, respectively. In vitro antibacterial assays revealed that E3f exerts synergistic effects with antibiotics and restores the susceptibility of clinically isolated MDR-GNB to antibiotics. The MIC was reduced to as low as 1/128 of the original value. In the murine intra-abdominal infection model, combined administration of E3f and meropenem markedly reduced bacterial loads in the liver and spleen compared with meropenem monotherapy. Meanwhile, the serum levels of pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 were decreased, indicating excellent in vivo efficacy. Acute toxicity tests demonstrated that E3f presents low toxicity and favorable overall safety in both Galleria mellonella larvae and mice. Furthermore, E3f effectively mitigated LPS-induced acute lung injury in mice by inhibiting ferroptosis, as evidenced by elevated FTH1 and GPX4 expression and reduced MDA levels in lung tissues. Collectively, these results confirm that E3f is a highly efficacious lead compound. Ebselen derivatives, which dually inhibit β-lactamases and produce ferroptosis-mediated anti-inflammatory activity, hold great promise as therapeutic agents against infections caused by MDR-GNB.

Infected wounds remain a critical clinical challenge, predominantly driven by Staphylococcus aureus (S. aureus, especially methicillin-resistant S. aureus, MRSA) biofilm formation, which triggers a vicious cycle of persistent infection, sustained inflammation, and progressive oxidative stress. Current clinical interventions fail to achieve simultaneous bacterial eradication, modulation of inflammation, and tissue regeneration, often with biocompatibility risks, whereas natural polysaccharide-based biomaterials offer a promising engineering strategy to address this dilemma. Herein, based on the nano-cocktail synthesis concept, we developed a caffeic acid (CA)-loaded nanocomposite (CA@ZSH) via green self-assembly from natural carbohydrate polymers (sodium alginate, SA; hyaluronic acid, HA) and zein for comprehensive infected wound therapy. CA was encapsulated in a homogeneous inner matrix of zein and bioactive polysaccharide HA, enabling stable drug loading/sustained release, along with HA-mediated regulation of inflammation and pro-regeneration. This core nanocomplex was further coated with polysaccharide SA as a hydrophilic shell to enhance colloidal stability and maintain a moist wound-healing microenvironment. CA@ZSH exhibited potent anti-S. aureus/MRSA activity via biofilm disruption, plus excellent free radical-scavenging, anti-inflammatory properties, and biocompatibility. In vivo, it markedly accelerated wound healing by upregulating CD31 and VEGF, thereby promoting wound closure, collagen deposition, and angiogenesis. This study provides a facile and robust polysaccharide-based nanotherapeutic engineering strategy for the management of wounds infected with drug-resistant bacteria.

Diabetic encephalopathy (DE) is a serious complication of diabetes mellitus characterized by progressive cognitive dysfunction, but its underlying mechanisms remain incompletely understood. Tau hyperphosphorylation and necroptosis are key pathological events in neurodegenerative diseases, but their roles in DE and the capacity of hydrogen sulfide (H2S) to regulate these processes have not been investigated. We tested whether H2S attenuates Tau hyperphosphorylation and necroptosis through the PI3K/AKT/GSK-3β signaling pathway to improve cognitive impairment in DE. In vitro, HT22 hippocampal neurons were exposed to high glucose (85 mM), and in vivo, a streptozotocin-induced diabetic mouse model was established. NaHS served as an exogenous H2S donor, and LY294002 was employed as a PI3K-specific inhibitor. Phosphoproteomic analysis revealed that high glucose suppressed PI3K/AKT/GSK-3β signaling and concurrently elevated Tau phosphorylation (p-Tau) and necroptosis markers (p-RIPK1, p-MLKL). NaHS treatment activated PI3K/AKT/GSK-3β signaling, reduced p-Tau, p-RIPK1, and p-MLKL levels, and normalized necroptotic morphology observed by transmission electron microscopy. Flow cytometry and lactate dehydrogenase release assays confirmed that NaHS attenuated high glucose-induced cell death. In diabetic mice, NaHS improved spatial learning and memory in the Morris water maze and novel object recognition tests, restored hippocampal CA1 neuron survival, and upregulated synaptic proteins (PSD95, SYP). Co-immunofluorescence demonstrated colocalization of p-Tau and p-MLKL in the hippocampal CA1 region, which was reduced by NaHS. All protective effects of NaHS were partially reversed by LY294002, establishing a PI3K-dependent mechanism. Network pharmacology identified 76 H2S-DE overlapping targets; machine learning ranked AKT family genes as top predictors. These findings demonstrate that H2S improves DE by activating PI3K/AKT/GSK-3β signaling, attenuating Tau hyperphosphorylation and necroptosis.

Diabetic encephalopathy (DE) is a significant complication of diabetes mellitus, primarily characterized by cognitive impairment. Pyroptosis-mediated neuronal pyroptosis is a key factor in diabetes-induced cognitive dysfunction. Research indicates that using NLRP3 inhibitors to block NLRP3 inflammasome activation or employing H2S to inhibit NF-κB activation can decrease pyroptosis and improve cognitive dysfunction in DE mice. Here, our aim is to design a bifunctional compound that can simultaneously inhibit the activation of NLRP3 and NF-κB to synergistically suppress neuronal pyroptosis for the clinical treatment of DE. This study reports the novel compound CYT-1, which significantly reduces levels of pyroptosis pathway proteins (p-NF-κB, NLRP3, Caspase-1, and GSDMD-N) and inflammatory factors (IL-1β and IL-18) in vitro. CYT-1 markedly increases H2S concentrations in both the serum and hippocampus of mice with DE. Western blotting and Nissl staining demonstrated that CYT-1 notably decreased pyroptosis-related protein levels and enhanced hippocampal neuron counts in vivo. The findings demonstrate that CYT-1 synergistically inhibits neuronal pyroptosis by concurrently suppressing NF-κB activation and NLRP3 inflammasome activity. This study may provide new ideas for drug research and clinical treatment of DE.

The pathogenesis of depression is associated with synaptic impairment and dysfunction in autophagy processes. Mendelian randomization (MR) analysis revealed that six GWAS IDs revealed a significant association between Beclin-1 levels and depression risk. Besides, all SNPs had a positive effect on depression risk. Analyzing neurons from depressed individuals using single-cell RNA sequencing (scRNA-seq) uncovered decreased expression of AKT, mTOR, and genes linked to synaptic plasticity. The activation of the PI3K/AKT/mTOR signaling has been demonstrated to control autophagy and have a protective effect on the nervous system. Hydrogen sulfide (H2S) is an endogenous gasotransmitter that can potentially treat various neurological disorders by improving neuronal synaptic plasticity. However, whether H2S regulates autophagy through PI3K/AKT/mTOR signaling, improves neuronal synaptic plasticity damage, and plays an antidepressant role is unclear. Our current research revealed that the reduction in the expression of p-PI3K, p-AKT, and p-mTOR proteins increase in neuronal autophagy activity and decline synaptic plasticity in mice with depression induced by chronic unpredictable mild stress (CUMS). Treatment with the exogenous hydrogen sulfide donor NaHS for one day and continuous treatment for one week improved the depression-like behaviors in the mice. Compared with those after one day of NaHS treatment, the above protein expression levels were restored and maintained, and the antidepressant effect was more significant after one week of continuous treatment with NaHS. Moreover, the PI3K inhibitor LY294002 was used to demonstrate that NaHS suppresses autophagy through activating the PI3K/AKT/mTOR signaling and ameliorates synaptic plasticity impairments. This study provides novel insights into the antidepressant mechanisms of H2S, highlighting its antidepressant therapeutic potential.

Nanoparticles (NPs) exhibit the potential to enhance plant tolerance to organic pollutant stress, but how they drive endogenous molecules to detoxify contaminants remains to be further investigated. This study clarified the modulatory mechanisms by which foliar or root application of biosynthesized titanium oxide NPs (g-nTiO2) alleviated atrazine (ATZ) toxicity to Lactuca sativa L. Compared with the ATZ-alone group, 10 mg/L g-nTiO2 intensified light-harvesting, photoelectron transfer, and reduced oxidative damage, thereby improving growth and inducing metabolic reprogramming. Specifically, g-nTiO2 activated pathways related to energy supply and defense detoxification, while stabilizing membrane lipid and nitrogen metabolism. Furthermore, the modulation of biomarkers involved in balancing cellular homeostasis and stimulating growth by g-nTiO2 ultimately boosted lettuce resistance to ATZ and physiological performance. Molecular docking analysis revealed that g-nTiO2 enhanced the Phase II metabolism of ATZ by glutathione and amino acids through increasing detoxification enzyme activities by 23-44%, which confirmed the driving role of NPs in alleviating ATZ phytotoxicity to lettuce. Collectively, these findings provide a prospective nanoenabled strategy for mitigating crop sensitivity to pesticide residues for safe and sustainable agricultural production.

Although advances in nanomedicine using nanoparticles (NPs) derived from natural compounds have provided us with much insight into how to recover from wound infections, exploring the wound healing pathway remains a new perspective that has attracted significant interest in addressing wound complications challenges. Here, we harnessed the therapeutic potential of novel selenium nanoparticles (SeNPs) derived from Purple sweet potato (PSp) extracts to promote wound healing through the regulation of pyroptosis-related pathways. PSp-SeNPs, with an average particle size of 80-100 nm, demonstrated significant antibacterial activity against S. aureus and MRSA clinical pathogens. The mechanism involves impairment of bacterial growth, biofilm formation, and metabolic processes through ATP depletion. Moreover, PSp-SeNPs impairs NLRP3-mediated pyroptosis including p-IκBα, p-NF-κB, IL-18, and IL-1β. This regulatory effect decreases inflammatory cytokines IL-6 and TNF-α while promoting angiogenesis and collagen formation through increased expression levels of TGF-β, VEGFA, and CD31, thus accelerating wound healing. In vivo assessments confirmed that PSp-SeNPs significantly enhanced wound healing without adverse effects, indicating their high biocompatibility and bioavailability. This groundbreaking study elucidates the therapeutic potential of PSp-based selenium nanoparticles, facilitating the development of precise and efficient treatment strategies for wound healing and diverse medical applications.

Background Diabetic cardiomyopathy (DCM) represents an important concern associated with diabetes, inhibiting pyroptosis has shown promising results in alleviating DCM symptoms. The objective of this work is to investigate the role and underlying mechanism of hydrogen sulfide (H2S) in the suppression of pyroptosis in the context of diabetic myocardial fibrosis (MF). Methods The effect of H2S on pyroptosis was detected using CCK-8, ELISA, and flow cytometry. The expression of Sestrin2 and its DNA methylation modification, as well as the quantification of pyroptosis-related proteins and the activation of the TLR4/MyD88/NF-κB signaling pathway, were measured using Western blot, ChIP, Immunofluorescence, and methylation-specific quantitative PCR. To establish a type 2 diabetes rat model, a high-fat diet was administered, followed by injection of streptozotocin (HFD/STZ). After five weeks, the rats received H2S treatment for four weeks, either with or without sh-Sestrin2. The effects of H2S treatment on myocardial function, tissue structure, and myocardial cell apoptosis were assessed. Furthermore, the CCK-8 assay was used to detect the cell viability induced by TGF-β. Cellular oxidative stress levels were measured by the ELISA method. Western blotting was applied to determine the protein expression levels of Collagen I, Fibronectin, α-SMA, NLRP3, caspase-1, GSDMD-N, and Sestrin2. Results H2S ameliorates HG-induced fibrosis, injury, pyroptosis, and inflammatory response of cardiac fibroblasts (CFs). H2S inhibits Sestrin2 methylation and up-regulates its expression through DNMT3a. H2S ameliorates HG-induced CFs pyroptosis and fibrosis through Sestrin2. Sestrin2 regulates HG-induced CFs pyroptosis and fibrosis through the TLR4/MyD88/NF-κB pathway. The myocardial tissue injury was improved after H2S treatment, and the results of the H2S treatment were reversed after knockdown of Sestrin2. In addition, TGF-β1 can induce an increase in the activity of cardiac fibroblasts and enhance cellular oxidative stress. Meanwhile, it significantly upregulates the expression of Collagen I, Fibronectin, α-SMA, as well as NLRP3, caspase-1, and GSDMD-N, and remarkably downregulates the expression of Sestrin2. However, H2S intervention can reverse the aforementioned phenomena. Conclusion H2S can suppress pyroptosis and ameliorate diabetic MF by inhibiting DNMT3a-mediated DNA methylation of Sestrin2 promoter through the TLR4/MyD88/NF-κB pathway.

Addressing the limitations of silver nanoparticles (AgNPs) and purple sweet potato anthocyanin extract (PSPAE) in infected wound management, this study developed a multifunctional nanoplatform (AgNPs-Cs-PSPAE) via green synthesis. Chitosan (Cs) served as a biocompatible carrier to co-anchor AgNPs and PSPAE, leveraging its cationic charge for enhanced biofilm penetration. In vitro characterization confirmed successful synthesis of spherical nanoparticles (similar to 173 nm) with high stability (Zeta potential +36.4 mV) and efficient PSPAE encapsulation (EE: 45.6 %, LC: 21.99 %). AgNPs-Cs-PSPAE demonstrated superior in vitro antibacterial efficacy against S. aureus (MIC: 128 mu g/mL), E. coli (MIC: 128 mu g/mL), and MRSA (MIC: 512 mu g/mL), significantly eradicating 83-90 % biofilms at 512 mu g/mL and scavenging ROS with 87 % H2O2 decomposition at 1 mg/mL. In vivo, treatment of S. aureus-infected murine wounds with AgNPs-Cs-PSPAE accelerated healing, achieving 76 % closure by day 7, reduced the bacterial burden by 4.8-fold, suppressed pro-inflammatory cytokines (IL-6, IL-1 beta, and TNF-alpha), and enhanced collagen deposition and angiogenesis, while upregulating TGF-beta, VEGF, and CD31. Safety assessments confirmed minimal toxicity. This Cs-assisted nanoplatform synergistically combines targeted antimicrobial action with antioxidant/anti-inflammatory activity, offering a promising strategy for wound healing in infected wounds.

Depression, characterized by a persistent low mood and apathy, is classified as a mental illness. Lipopolysaccharide (LPS), an inflammatory inducer, reduces plasma concentrations of hydrogen sulfide (H2S) and gamma-aminobutyric acid (GABA) in mice, resulting in depressive-like behaviors. H2S, an endogenous gaseous signaling molecule, is crucial for maintaining normal physiological functions of the central nervous system. GABA, an inhibitory neurotransmitter, has been demonstrated to mitigate depression-like behaviors in mice subjected to chronic stress. Sodium hydrosulfide (NaHS), an H2S donor, alleviates LPS-induced depressive-like behaviors in mice; however, its rapid release of H2S may lead to accumulation and potential toxicity. This study aimed to mimic the body's natural slow production of H2S and GABA. To this end, three novel multifunctional donors─BGS, BGF, and BGA─were designed and synthesized. Among them, BGS showed reduced toxicity to HT-22 cells and a sustained release profile in vitro. Furthermore, BGS increased plasma levels of H2S and GABA in mice, ameliorated LPS-induced depressive-like behaviors, enhanced neuronal count in the hippocampal CA1 subregion, decreased p-NF-κB levels, and upregulated the expression of synaptic proteins SYN and PSD-95. These results suggest that BGS not only elevates plasma H2S and GABA levels but also inhibits NF-κB activation, enhances synaptic protein expression, and improves synaptic plasticity, thus exerting a multifaceted antidepressant effect.

Abstract Diabetic encephalopathy (DE) is a severe complication of the central nervous system associated with diabetes. This research aims to investigate the regulatory role of mammalian target of rapamycin (mTOR) on nuclear factor kappa-B (NF-κB) in mice with DE, and the neuroprotective effect and therapeutic mechanisms of luteolin, a natural flavonoid compound with anti-inflammatory, antioxidant, and neuroprotective properties. The results indicated that treatment with luteolin improved the degree of cognitive impairment in mice with DE. It also decreased the levels of p-mTOR, p-NF-κB and histone deacetylase 2 (HDAC2) and increased the expression of brain-derived neurotrophic factor (BDNF) and synaptic-related proteins. Furthermore, protein-protein interaction (PPI) and the Gene Ontology (GO) analysis revealed that luteolin was involved in the regulatory network of HDAC2 expression through the mTOR/NF-κB signaling cascade. Our bioinformatics and molecular docking results indicated that luteolin may also directly target HDAC2, as an HDAC2 inhibitors, to alleviate DE, complementing mTOR/NF-κB signaling inhibition. Analysis of luteolin's target genes and their interactions suggested effect on HDAC2 and cognition. In conclusion, HDAC2 and tau hyperphosphorylation are regulated by the mTOR/NF-κB signaling cascade in DE, and luteolin is found to reverse these effects, demonstrating its protective role in DE. Article Highlights • Luteolin's neuroprotective effect on diabetic encephalopathy, targeting mTOR/NF-κB signaling. • Demonstrating decreases in p-mTOR, p-NF-κB, and HDAC2, and increases in BDNF and synaptic proteins. • Highlighting Luteolin's potential direct targeting to HDAC2, in order to provide a new approach for DE treatment.

The ecological safety of copper oxide nanoparticles (CuO NPs) in the environment determines the advancement of nano-agriculture owing to breakthroughs in nanotechnology; however, the release of Cu2+ is an uncontrollable factor. Currently, the trade-off mechanisms of CuO NPs and Cu2+ dominating the potential hazards of plant-nano systems remain unclear. This study proposed the trade-off strategy for reconstructing physiological responses and metabolic profiles and deciphered the differential regulation of dominant CuO NPs and Cu2+ in plants. The results showed that 100 and 500 mg/kg CuO NPs promoted root fresh weight but reduced shoot fresh weight, while 1000 mg/kg Cu2+ demonstrated the strongest inhibition on both roots and shoots. The net photosynthetic perturbation in photosynthetic disorders is accompanied by superoxide anion and hydrogen peroxide accumulation, which are severe under 1000 mg/kg CuO NPs and Cu2+ stress. Metabolomics revealed that CuO NPs significantly altered coumaric acid and derivatives, for example, down-regulating coumaroyl hexoside (isomers of 690 and 691) by 40.79 %. Additionally, Cu2+ treatment severely interfered with the dominant metabolic response, activating plant hormone signal transduction and α-linolenic acid metabolism. The trade-off strategies of galactose metabolism, amino sugar and nucleotide sugar metabolism, pantothenate and coenzyme A (CoA) biosynthesis, and β-alanine metabolism as differential metabolism were confirmed by comparing the CuO NPs and Cu2+ exposure. Protein secondary structure analysis revealed specific regulation of protein conformation upon exposure to CuO NPs and Cu2+. These findings provide new insights into differential metabolism and environmental effects in plant-nano systems.

Damaged skin is susceptible to invasion by harmful microorganisms, especially Staphylococcus aureus and Escherichia coli, which can delay healing. Epigallocatechin-3-gallate (EGCG) is a natural compound known for effectively promoting wound healing and its potent anti-inflammatory effects. However, its application is limited due to its susceptibility to oxidation and isomerization, which alter its structure. The use of zeolitic imidazolate framework-8 (ZIF-8) can effectively tackle these issues. This study introduces an oxygen (O-2) and hydrogen peroxide (H2O2) self-supplying ZIF-8 nanoplatform designed to enhance the bioavailability of EGCG, combining photodynamic therapy (PDT) and chemodynamic therapy (CDT) to improve antibacterial properties and ultimately accelerate wound healing. For this purpose, EGCG and indocyanine green (ICG), a photosensitizer, were successively integrated into a ZIF-8, and coated with bovine serum albumin (BSA) to enhance biocompatibility. The outer layer of this construct was further modified with manganese dioxide (MnO2) to promote CDT and calcium peroxide (CaO2) to supply H2O2 and O-2, resulting in the final nanoplatform EGCG-ICG@ZIF-8/BSA-MnO2/CaO2 (EIZBMC). In in vitro experiments under 808 nm laser, EIZBMC exhibited synergistic antibacterial effects through PDT and CDT. This combination effectively released reactive oxygen species (ROS), which mediated oxidative stress to inhibit the bacteria. Subsequently, in a murine model of wound infection, EIZBMC not only exerted antibacterial effects through PDT and CDT but also alleviated the inflammatory condition and promoted the regeneration of collagen fibers, which led to accelerated wound healing. Overall, this research presents a promising approach to enhancing the therapeutic efficacy of EGCG by leveraging the synergistic antibacterial effects of PDT and CDT. This multifunctional nanoplatform maximizes EGCG's anti-inflammatory properties, offering a potent solution for promoting infected wound healing.

In recent years, contamination of aquatic systems with Avermectin (AVM) has emerged as a significant concern. This contamination poses substantial challenges to freshwater aquaculture. Plant-derived Quercetin (QUE), known for its anti-inflammatory, antioxidant, and ferroptosis-inhibiting properties, is commonly employed as a supplement in animal feed. However, its protective role against chronic renal injury in freshwater carp induced by AVM remains unclear. This study assesses the influence of dietary supplementation with QUE on the consequences of chronic AVM exposure on carp renal function. The carp were subjected to a 30-day exposure to AVM and were provided with a diet containing 400 mg/kg of QUE. Pathological observations indicated that QUE alleviated renal tissue structural damage caused by AVM. RT-QPCR study revealed that QUE effectively reduced the increased expression levels of pro-inflammatory factors mRNA produced by AVM exposure, by concurrently raising the mRNA expression level of the anti-inflammatory factor. Quantitative analysis using DHE tests and biochemical analysis demonstrated that QUE effectively reduced the buildup of ROS in the renal tissues of carp, activity of antioxidant enzymes CAT, SOD, and GSH-px, which were inhibited by AVM, and increased the content of GSH, which was induced by prolonged exposure to AVM. QUE also reduced the levels of MDA, a marker of oxidative damage. Furthermore, assays for ferroptosis markers indicated that QUE increased the mRNA expression levels of gpx4 and slc7a11, which were reduced due to AVM induction, and it caused a reduction in the mRNA expression levels of ftl, ncoa4, and cox2, along with a drop in the Fe2+ concentration. In summary, QUE mitigates chronic AVM exposure-induced renal inflammation in carp by inhibiting the transcription of pro-inflammatory cytokines. By blocking ROS accumulation, renal redox homeostasis is restored, thereby inhibiting renal inflammation and ferroptosis. This provides a theoretical basis for the development of freshwater carp feed formula.

It is unclear whether the parent Saxagliptin (SAX) in vivo is the same as that in vitro, which is twice that of 5-hydroxy Saxagliptin (5-OH SAX). This study is to construct a Pharmacokinetic-Pharmacodynamic (PK-PD) link model to evaluate the genuine relationship between the concentration of parent SAX in vivo and the effect. First, we established a reliable Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS/MS) method and DPP-4 inhibition ratio determination method. Then, the T2DM rats were randomly divided into four groups, intravenous injection of 5-OH SAX (0.5 mg/kg) and saline group, intragastric administration of SAX (10 mg/kg) and Sodium carboxymethyl cellulose (CMC-Na) group. Plasma samples were collected at different time points for subsequent testing. Finally, we used the measured concentrations and inhibition ratios to construct a PK-PD link model for 5-OH SAX and parent SAX. A two-compartment with additive model showed the pharmacokinetic process of SAX and 5-OH SAX, the concentration-effect relationship was represented by a sigmoidal Emax model and sigmoidal Emax with E0 model for SAX and 5-OH SAX, respectively. Fitting parameters showed SAX was rapidly absorbed after administration (Tmax=0.11 h, t1/2, ka=0.07 h), widely distributed in the body (V ≈ 20 L/kg), plasma exposure reached 3282.06 ng*h/mL, and the elimination half-life was 6.13 h. The maximum plasma dipeptidyl peptidase IV (DPP-4) inhibition ratio of parent SAX was 71.47

Parkinson’s disease (PD) is a severe neurodegenerative disease associated with the loss of dopaminergic (DA) neurons in the substantia nigra (SN). Although its pathogenesis remains unclear, microglia-mediated neuroinflammation significantly contributes to the development of PD. Here we showed that the sine oculis homeobox (SIX) homologue family transcription factors SIX2 exerted significant effects on neuroinflammation. The SIX2 protein, which is silenced during development, was reactivated in lipopolysaccharide (LPS)-treated microglia. The reactivated SIX2 in microglia mitigated the LPS induced inflammatory effects, and then reduced the toxic effect of conditioned media (CM) of microglia on co-cultured MES23.5 DA cells. Using the LPS-stimulated Cx3cr1-CreERT2 mouse model, we also demonstrated that the highly-expressed SIX2 in microglia obviously attenuated neuroinflammation and protected the DA neurons in SN. Further RNA-Seq analysis on the inflammatory activated microglia revealed that the SIX2 exerted these effects via up-regulating the FXYD domain containing ion transport regulator 2 (FXYD2). Taken together, our study demonstrated that SIX2 was an endogenous anti-inflammatory factor in microglia, and it exerted anti-neuroinflammatory effects by regulating the expression of FXYD2, which provides new ideas for anti-neuroinflammation in PD.

Hydrogen sulfide (H2S) is a crucial endogenous gas signaling molecule in the central nervous system, playing a significant role in anti-inflammation, anti-oxidation, and neuroprotection. Our studies have shown that the level of H2S is closely related to hypothalamic inflammation-induced obesity. Compared to neurons, astrocytes, exhibit a preferential activation in response to high-fat diet-induced hypothalamic inflammation. Whether the reduction of hydrogen sulfide in astrocytes is the cause of neuronal damage leading to hypothalamic inflammation is worth exploring. The key to discussing the role of astrocytes is to accurately and quickly detect the levels of H2S in the complex microenvironment of the brain. In this work, we designed and synthesized a H2S near-infrared fluorescence probe (NIR-HS) based on dicyanideisophorone. NIR-HS selectively responds to H2S at 668 nm, with an effective linear range (0-20 mu M), high sensitivity (35 nM). This probe has been successfully applied to detect hydrogen sulfide levels in astrocytes and to perform H2S fluorescence imaging on the slices of the ventromedial nucleus of the hypothalamus. Additionally, the effect of the H2S donor drug allicin on the levels of H2S in the hypothalamus and its therapeutic effect on hypothalamic inflammation were confirmed using NIR-HS and a series of signal pathway studies. Ultimately, we revealed the mechanism of that H2S inhibits hypothalamic neuronal inflammation by suppressing the expression of the pro-inflammatory factor IL-1 beta in astrocytes.

PURPOSE:Clear cell renal cell carcinomas (ccRCCs) are the most common form of renal cancer in the world. The loss of extracellular matrix (ECM) stimulates cell apoptosis, known as anoikis. A resistance to anoikis in cancer cells is believed to contribute to tumor malignancy, particularly metastasis; however, the potential influence of anoikis on the prognosis of ccRCC patients is not fully understood. METHODS:In this study, anoikis-related genes (ARGs) with discrepant expression were selected from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. The anoikis-related gene signature (ARS) was built using a combination of the univariate Cox and least absolute shrinkage and selection operator (LASSO) analyses. ARS was also evaluated for their prognostic value. We explored the tumor microenvironment and enrichment pathways between different clusters of ccRCC. We also examined differences in clinical characteristics, immune cell infiltration and drug sensitivity between the high- and low-risk sets. In addition, we utilized three external databases and quantitative real-time polymerase chain reaction (qRT-PCR) to validate the expression and prognosis of ARGs. RESULTS:Eight ARGs (PLAUR, HMCN1, CDKN2A, BID, GLI2, PLG, PRKCQ and IRF6) were identified as anoikis-related prognostic factors. According to Kaplan-Meier (KM) analysis, ccRCC patients with high-risk ARGs have a worse prognosis. The risk score was found to be a significant independent prognostic indicator. According to tumor microenvironment (TME) scores, stromal score, immune score, and estimated score of the high-risk group were superior to those of the low-risk group. There were significant differences between the two groups regarding the amount of infiltrated immune cells, immune checkpoint expression as well as drug sensitivity. A nomogram was constructed using ccRCC clinical features and risk scores. The signature and the nomogram both performed well in predicting overall survival (OS) for ccRCC patients. According to a decision curve analysis (DCA), clinical treatment options for patients with ccRCC could be improved using this model. CONCLUSION:The results of validation from external databases and qRT-PCR were basically agreement with findings in TCGA and GEO databases. The ARS serving as biomarkers may provide an important reference for individual therapy of ccRCC patients.

Background and Purpose：Hypoxia, as a risk factor for pulmonary hypertension (PH), was an inducing factor for pulmonary artery endothelial cells (PAECs) injury and inflammation. Pyroptosis induced cell death along with the maturation and secretion of the inflammatory mediators. However, the correlation among pyroptosis, PAECs injury and inflammation remain unknown. Here, we explored, in detail, the effect of hypoxia on pyroptosis of PAECs. Experimental approach：Using RNA-seq sequencing method, we screened differentially expressed genes in pulmonary artery in SU5416-induced hypoxia PH model. Next, we verified the role and mechanism of the differentially expressed gene GPR146 in PAECs by immunohistochemistry, immunofluorescence, CCK8, western blotting, realtime PCR, and LDH release experiments. Key Results: Our results showed that GPR146 was highly expressed in PH human lung tissue and Sugen5416/hypoxia (SuHx) induced rat PAH lung tissues. Meanwhile, our data suggested the expression of pyroptosis-related proteins was remarkably increased under hypoxia both in vivo and in vitro, which were inhibited by silencing GPR146. Moreover, inhibiting NLRP3 or caspase-1 effectively suppressed cleavage of caspase-1, production of interleukin (IL)-1β and IL-18 in PAECs by hypoxia and overexpression of GPR146. Conclusion and Implications: Our results indicated that GPR146 induced pyroptosis and inflammatory responses through the NLRP3/caspase-1 signaling axis, triggering endothelial injury and vascular remodeling. Hypoxia could promote PAECs pyroptosis through upregulation of GPR146 to affect the progression of PH, which might provide novel targets for treatment of PH.