Di-(2-ethylhexyl) phthalate (DEHP), a ubiquitous environmental plasticizer, has been increasingly linked to the exacerbation of inflammatory skin conditions, especially psoriasis. However, the mechanisms and effective therapeutic strategies targeting DEHP-aggravated psoriasis remain elusive. We integrated network toxicology, network pharmacology, and molecular docking to explore the targets of DEHP-exacerbated psoriasis and the protective effects of shikonin (SH). These findings were validated in a DEHP/IMQ-induced mouse model using a novel shikonin nanoparticle (SH-NP) that overcomes SH's inherent hydrophobicity. Computational analyses revealed that SH counteracts DEHP skin toxicity through a multi-target network, identifying the p38 mitogen-activated protein kinase (p38 MAPK), TNF, IL-1β, proliferating cell nuclear antigen (PCNA), and matrix metalloproteinases 2 and 9 (MMP2/9) as core therapeutic nodes. Molecular docking verified this network, revealing robust binding between SH and key targets with binding energies ranging from -6.0 to -7.8 kcal/mol, consistently outperforming DEHP. In vivo experiments demonstrated that topical application of SH-NP significantly ameliorated macroscopic skin injury and reduced PASI scores. Mechanistically, SH-NP effectively reversed the DEHP-exacerbated inflammatory microenvironment by decreasing macrophage and mast cell infiltration in the dermis, reducing pro-inflammatory cytokine levels, and inhibiting the phosphorylation of p38 MAPK. Furthermore, SH-NP treatment successfully suppressed keratinocyte hyperproliferation, as evidenced by downregulated PCNA expression and reduced epidermal thickness. SH-NPs also markedly downregulated the elevated levels of MMP2/9 in mice, thereby mitigating collagen degradation and maintaining dermal matrix integrity. This study shows that SH-NPs alleviate DEHP-aggravated psoriasis by suppressing local inflammation, keratinocyte hyperproliferation, and collagen degradation, offering a targeted nanostrategy for environmentally exacerbated skin diseases.

Fine particulate matter (PM2.5) is an environmental factor that triggers gastrointestinal diseases. However, the effects of PM2.5 on intestinal function are not fully understood. This study established an environmental exposure cell model to explore PM2.5-induced intestinal permeability alteration and its mechanisms. Intestinal barrier permeability was evaluated via trans-epithelial electrical resistance (TEER) measurement and FITC–dextran paracellular penetration analysis, followed by detection of intercellular junction protein β-catenin and its coding gene CTNNB1. Expression of inflammatory cytokines (TNF-α, IL-6) and phosphorylation of PI3K and AKT were assessed using quantitative real-time polymerase chain reaction and Western blot, respectively. Reactive oxygen species (ROS) and malondialdehyde were measured using commercial kits to observe cellular oxidative stress. The results showed that PM2.5 impaired the intestinal barrier, as indicated by reduced TEER, increased FITC–dextran penetration, down-regulated expression of β-catenin and CTNNB1. Additionally, compared with the control, inflammatory cytokines and oxidative stress markers were significantly elevated after PM2.5 exposure. The ratio of p-PI3K/PI3K and p-AKT/AKT was also up-regulated in PM2.5-exposed Caco-2 cells. Pretreatment with PI3K inhibitor LY294002 and ROS scavenger NAC modulated β-catenin expression, reduced inflammation/ROS, and alleviated the hyperpermeability of Caco-2 cells. Thus, our results reveal that PM2.5 induces PI3K/AKT-mediated inflammation and ROS generation in Caco-2 cells, leading to intestinal barrier impairment.

BACKGROUND: Epithelioid glioblastoma (GBM) is characterized by its highly aggressive behavior and the presence of the BRAF V600E mutation. However, the impact and underlying mechanisms of the BRAF V600E mutation on GBM stemness and invasiveness are unknown. Dysregulation of N6-methyladenosine (m6A) modification is closely associated with the progression of various cancers. The role of m6A modification in BRAF V600E-mutant GBM has not been defined. METHODS: Functional assays were performed to evaluate the impact of BRAF V600E on stemness phenotypes of glioma stem-like cells (GSCs) and invasive phenotypes of GBM cells in vitro and in vivo. Mechanistic investigations involved m6A quantification, qPCR, western blotting, co-immunoprecipitation, MeRIP-seq, luciferase reporter assays, and transmission electron microscopy to elucidate the mechanism by which BRAF V600E regulates stemness and invasiveness. These findings were further supported by evidence from public GBM patient databases and tumor samples. RESULTS: We found that BRAF V600E significantly upregulated METTL3 expression via ERK signaling in GSCs and GBM cells, thereby promoting stemness and invasiveness. METTL3 established a positive feedback loop with BRAF V600E to facilitate m6A modification enrichment, thereby inducing autophagy. The BRAF V600E-driving stemness and invasiveness were autophagy-dependent. In vivo experiments showed that BRAF V600E-expressing GBM was responsive to both the METTL3 inhibitor STM2457 and the autophagy inhibitor HCQ. CONCLUSIONS: This study reveals that the BRAF V600E/MEK/ERK/METTL3 positive feedback loop promotes autophagy, driving the stemness of GSCs and the invasiveness of GBM cells via m6A modification in vitro and in vivo. Our results suggest that METTL3 and autophagy are promising therapeutic targets in BRAF V600E-mutant GBM.

Exposure to fine particulate matter (PM2.5) poses a major threat to skin health, yet effective prevention strategies remain limited. Shikonin, a naphthoquinone derived from Lithospermum erythrorhizon, exhibits potent antioxidant and anti-inflammatory activities. However, its therapeutic application is limited by low bioavailability. To address this limitation, we developed shikonin-loaded nanoparticles (SH-NPs) using an emulsion solvent evaporation method and characterized their physicochemical properties. The protective effects of SH-NPs against PM2.5-induced skin damage were evaluated in a mouse model. The SH-NPs exhibited favorable characteristics, including a mean particle size of 209.03 ± 2.45 nm, a PDI of 0.064 ± 0.03, and a zeta potential of –17.69 ± 2.06 mV. The encapsulation efficiency is 88% and the drug loading capacity is 5.5%, respectively. In vitro, SH-NPs significantly enhanced cellular uptake in HaCaT cells. In vivo, treatment with SH-NPs significantly improved skin structural disorders, epidermal thickening, and collagen fiber reduction, while downregulating the expression of MMP-2 and MMP-9. Furthermore, SH-NPs increased the expression of SOD1 and SOD2, reduced MDA levels, and decreased the expression of TNF-α, IL-1β, and NO. In conclusion, SH-NPs attenuated PM2.5-induced skin toxicity via enhanced antioxidant, anti-inflammatory, and anti-degradation mechanisms, offering a novel strategy to boost shikonin bioavailability and prevent PM2.5-related skin damage.

Epidemiological studies prove that type II diabetes, characterized by insulin resistance (IR), may be caused by fine particulate matter 2.5 (PM2.5). However, underlying mechanisms whereby PM2.5, particularly during short-term exposure, induces liver dysfunction leading to IR remains poorly understood. In the present study, HepG2 cells and the BALB/c mouse model were used to explore how PM2.5 affects insulin sensitivity. The effects of subacute PM2.5 exposure on glucose metabolism were examined using commercial kits. Oxidative stress and inflammation were detected by fluorescent staining and RT-qPCR. The phosphorylation of PI3K/AKT was examined by Western blot. Subacute PM2.5 exposure induced IR, as reflected by increased glucose levels in cell supernatants, elevated insulin levels, and the impaired intraperitoneal glucose tolerance test in mice. PM2.5 induced oxidative stress, as evidenced by increased reactive oxygen species, cytochrome P450 2E1, and malondialdehyde, along with reduced superoxide dismutase 1/2 and silent information regulator 1. IL-6 and TNF-α were found to be upregulated using RT-qPCR. Western blot showed that PM2.5 inhibited the PI3K-AKT signaling pathway, indicated by the decreased phosphorylation of PI3K/AKT in HepG2 cells. Additionally, H&E staining showed only mild hepatic injury in mice liver. These results firmly suggest that subacute PM2.5 exposure induces insulin resistance through oxidative stress, inflammation, and the inhibition of the PI3K-AKT signaling pathway.

Background Pancreatic cancer is one of the most aggressive and deadly malignancies of the digestive system, with an extremely low five-year survival rate, making treatment highly challenging. Liquid-liquid phase separation (LLPS) is a mechanism that regulates the dynamic aggregation of intracellular proteins and nucleic acids. In cancer, tumor-associated proteins and gene mutations can regulate the formation of LLPS, influencing the quantity and functionality of intracellular condensates, thereby promoting abnormal cellular behavior and contributing to tumorigenesis and cancer progression. Although previous studies have suggested that LLPS may be involved in cancer progression by affecting mechanisms such as stress response and protein aggregation, its specific role in pancreatic cancer remains unclear. Therefore, exploring the molecular mechanisms of LLPS in pancreatic cancer could uncover new biomarkers and potential therapeutic targets, facilitating the development of personalized treatment strategies.Methods LLPS-related signatures were obtained from the TCGA database and Gene Cards. Over 100 machine learning methods were applied to screen for prognostic gene sets closely related to survival, followed by the construction of a risk model. External validation of the model was performed using the GEO database. Single-cell RNA sequencing analysis was conducted on the GSE155698 dataset to assess gene expression profiles and risk scores.Result A total of 55 LLPS-related prognostic genes were identified in pancreatic cancer. Through multiple machine learning methods, 11 key prognostic genes (CKB, PRSS3, KRT6A, DLGAP5, EPHA2, FAM83B, FOXM1, IGF2BP3, KRT16, CASP14, and TOP2A.) were selected, demonstrating high sensitivity and specificity as biomarkers for the diagnosis and prognosis of PAAD patients. The risk score signature constructed based on machine learning was found to be an independent factor associated with high mortality, advanced clinicopathological features, and chemotherapy resistance. Furthermore, this signature was closely related to lipid biosynthesis pathways, cell cycle-related pathways, and KRAS/TP53 mutation profiles, suggesting its potential role in enhancing the number and functionality of abnormal lipid-based "droplets" and accelerating cell cycle progression, thereby driving pancreatic cancer development. Additionally, the signature was strongly associated with immune-suppressive cell infiltration and immune checkpoint expression, indicating its role as a key immunosuppressive factor in the tumor microenvironment. Single-cell RNA sequencing revealed that PKP3, EPHA2, and KRT16 were specifically expressed in pancreatic ductal epithelial cells.Conclusions This study provides insights into the potential links between LLPS-related molecular characteristics and clinical features, the tumor microenvironment, and clinical drug response. It highlights the crucial role of LLPS in pancreatic cancer progression and treatment resistance, offering valuable new perspectives for the prognosis and treatment response prediction in PAAD.

Background: To analyze the current status, hotspots, and cutting-edge trends of PM2.5 health effects of PM2.5, using CiteSpace. Methods: The Web of Science Core Collection Science Citation Index Expanded was searched for relevant articles from January 1, 2013, to December 31, 2023. Network maps identifying authors, institutions, countries, keywords, co-cited authors, journals, references, and research trends were then created using CiteSpace. Results: A total of 2174 articles on the health effects of PM2.5 were identified between 2013 and 2023, with an increasing trend in annual publications. The United States had the highest number of articles on this topic, followed by China. The Chinese Academy of Sciences (CAS) is the leading institute in terms of paper production. Hoek was the most prolific author, focusing on the health consequences of air pollution. Among referenced journals, ENVIRON HEALTH PERSP ranked first, while Pope Ca was the most cited author. Current research focuses on reactive oxygen species (ROS), inflammation, oxidative stress, cardiovascular and respiratory diseases, PM2.5 sources, and chemical composition. The field is currently experiencing a phase of rapid expansion. Conclusion: The findings of this bibliometric analysis offer insight into the status and direction of research on PM2.5 and its health impacts, which aid scientists in coming up with new directions for their investigation.

Exposure to airborne fine particulate matter (PM 2.5 ) is strongly associated with poor fertility and ovarian damage. However, the mechanism underlying this remains largely unclear. Here, we found that PM 2.5 markedly impaired murine ovarian reserve, decreased hormone levels, and aggravated ovarian inflammation. Circulating interleukin-6 (IL-6) was elevated in PM 2.5 -exposed mice and was further confirmed to mediate this damage by IL-6 recombinant protein intervention. PM 2.5 exposure led to increased alveolar macrophage infiltration in the lungs. However, alveolar macrophage clearance with clodronate liposomes could not fully reverse the elevated IL-6 levels and ovarian injury, suggesting that alveolar macrophages were probably not the only source of circulating IL-6. Further experiments indicated that IL-6 mainly targeted ovarian theca–interstitial cells and impaired testosterone synthesis via suppressing the peroxisome proliferator-activated receptor γ (PPARγ) pathway. In addition, apoptosis of granulosa cells and restriction of follicular growth were observed in co-cultures with IL-6-treated theca–interstitial cells, which could be further reversed by the PPARγ agonist. Moreover, IL-6-neutralizing antibodies ameliorated PM 2.5 -induced ovarian damage. Notably, increased levels of circulating IL-6 were observed in premature ovarian aging patients and were inversely associated with their ovarian function. In summary, our findings offer a mechanistic explanation for PM 2.5 -induced ovarian dysfunction and verify IL-6 as a biomarker and potential therapeutic target.

Chronic elevated free fatty acids (FFAs) impair pancreatic β cells, but the mechanisms remain elusive. In this study, palmitic acid (PA) impaired viability and glucose-stimulated insulin secretion of INS-1 cells. Microarray analysis showed that PA markedly altered the expression of 277 probe sets of genes with 232 upregulated and 45 downregulated (fold change ≥ 2.0 or ≤ -2.0; P < 0.05). Gene Ontology analysis displayed a series of the biological process of the differentially expressed genes, such as intrinsic apoptotic signaling pathway in response to endoplasmic reticulum (ER) stress and oxidative stress, inflammatory response, positive regulation of macroautophagy, regulation of insulin secretion, cell proliferation and cycle, fatty acid metabolic process, glucose metabolic process and so on. Kyoto Encyclopedia of Genes and Genomes analysis demonstrated molecular pathways with which the differentially expressed genes associated, including NOD-like receptor, NF-κB and PI3K-Akt signaling pathways, apoptosis, adipocytokine signaling pathway, ferroptosis, protein processing in ER, fatty acid biosynthesis and cell cycle. Moreover, PA promoted protein expression of CHOP, cleaved caspase-3, microtubule-associated proteins light chain 3 (LC3)-II, NOD-like receptor pyrin domain containing 3 (NLRP3), cleaved IL-1β and Lcn2, increased reactive oxygen species, apoptosis and the ratio of LC3-II/I, and reduced p62 protein expression, intracellular glutathione peroxidase and catalase levels, suggesting activation of ER stress, oxidative stress, autophagy and NLRP3 inflammasome. The results indicate the impaired role of PA and the global gene expression profile of INS-1 cells following PA intervention, providing new insights into the mechanisms involving the damage of pancreatic β cells by FFAs.

Liver fibrosis has the potential to progress into liver cirrhosis, liver failure, and even death. Hepatic stellate cells (HSCs) activation play a central role in liver fibrosis, and persistently damaged hepatocytes secrete soluble factors that activate transdifferentiation of HSCs into myofibroblasts. Our previous studies indicated that fine particulate matter (PM2.5) can activate HSCs by stimulating hepatocytes to secrete TGF-β1. However, whether PM2.5 activates HSCs by regulating oxidative stress in hepatocytes remains uncertain. Oleanolic acid (OA) has been widely used in the clinic for hepatoprotection in Chinese medicine. In the present study, OA-loaded nanoparticles (OA-NP) with high solubility were used to attenuate the activation of HSCs induced by PM2.5-treated hepatocytes, and further studies were performed to explore the mechanism in which OA-NP plays a vital part. Our results showed that consistently PM2.5 treatment induced oxidative stress in hepatocytes. Moreover, the activation of HSCs induced by PM2.5-treated hepatocytes was reversed by antioxidant N-acetylcysteine treatment. Hence, PM2.5 may participate in the activation of HSCs by regulating oxidative stress in hepatocytes. Using a co-cultivation system, our results proved pretreatment with OA-NP significantly attenuates the activation of HSCs induced by PM2.5-exposed hepatocytes. In addition, the TGF-β1 expression and oxidative stress in hepatocytes with PM2.5 treated were reduced by the incubation with OA-NP. These observations demonstrated that OA-NP protects against the activation of HSCs by decreasing the TGF-β1 level and oxidative stress in PM2.5-exposed hepatocytes.

The present study was to investigate the inhibitory effect and underlying mechanism of Tormentic acid (TA) on hepatic stellate cells (HSCs). HSC-T6 cells were stimulated with Platelet-derived growth factor-BB (PDGF-BB) and TA, and then cell proliferation, apoptosis, inflammatory factor, and collagen-related indicators were detected. In order to elucidate the potential mechanism, the PI3K/Akt/mTOR and NF-kappa B signalling pathways were also detected. The results showed that TA treatment markedly inhibited PDGF-BB-stimulated HSC-T6 cell activation, as evidenced by the inhibition of cell proliferation, migration and colony formation, as well as the decreased expression of TGF-beta and alpha-SMA. TA treatment caused a significant increase in the activity of lactate dehydrogenase and significantly promoted cell apoptosis. TA treatment significantly reduced aspartate aminotransferase, alanine aminotransferase and total bilirubin activity. Importantly, TA inhibited the expression of collagen type I and III, alleviating the excessive deposition of extracellular matrix (ECM). Further experiments showed that TA administration significantly inhibited the phosphorylation of PI3K, Akt, FAK and mTOR and the protein expression of P70S6K, indicating the inhibition of the PI3K/Akt/mTOR pathway. Moreover, treatment with TA markedly decreased the phosphorylation of I kappa B alpha, NF-kappa B p65 and IKK alpha/beta, thereby blocking the NF-kappa B signal transduction. In summary, this study demonstrates that TA significantly inhibits HSC activation and promotes cell apoptosis via the inhibition of the PI3K/Akt/mTOR and NF-kappa B signalling pathways. Significance of the study Tormentic acid (TA) could inhibit HSC activation and alleviate collagen-based ECM deposition, suggesting that TA exerted anti-hepatic fibrosis. Further mechanism research revealed that the inhibition of TA on HSC activation might be through blocking the PI3K/Akt/mTOR and NF-kappa B signalling pathways. These findings provided a new cue to understand the protective effect of TA against liver fibrosis, which may provide a potential nature medicine for the treatment of liver fibrosis.

Background: IL-35 and IL-35 induced IL-35-producing regulatory T cells (iTr35) have been reported to inhibit Th2 response in allergic rhinitis (AR). However, the effect of IL-35 and iTr35 on type II innate lymphoid cells (ILC2) in AR is not well characterized.Objective: We aimed to explore the effect of IL-35 and iTr35 on type II innate lymphoid cells (ILC2) in AR.Methods: Twenty-five AR patients and 20 healthy controls were recruited. The proteint expression of IL-35, the proportion of ILC2, IL-4+ILC2, IL-5+ILC2, IL-13+ILC2 and IL-35+CD4+CD25+ cells were detected by enzyme-linked immunosorbnent assay (ELISA) and flow cytometry. The expression and regulation of IL-35 receptor in ILC2 were analyzed by real-time PCR. The effect of IL-35 on ILC2 differentiation and cytokine production were analyzed by real-time PCR and ELISA. The iTr35 were cocultured with ILC2 in Transwell or non-Transwell system to explore the regulation of the iTr35 on ILC2 differentiation and function using RT-PCR and ELISA. AR mice models were also established to confirm the role of IL-35 in the regulation of ILC2 in vivo.Results: AR patients had decreased IL-35 expression and iTr35 proportion and increased proportion of ILC2 and type II cytokines compared with controls (P<0.05). The recombinant IL-35 inhibited ILC2 differentiation and type II cytokine production through GATA3/RORα pathway by regulation of IL-12Rβ2 and gp130. IL-35 promoted the inducible costimulatory molecule (ICOS) expression by iTr35 and ICOS ligand (ICOSL) expression by ILC2. iTr35 inhibited the function of ILC2s through ICOS:ICOS ligand. IL-35 treated AR mice presented as inhibited ILC2 inflammation compared with control mice.Conclusions: IL-35 inhibited ILC2 responses directly or through mutual contaction between iTr35 and ILC2 in AR, suggesting that IL-35 may be used as potential treatment target in AR.Funding: This study was supported by grants from the National Natural Science Grant of China (No.81600785, No. 81700892, No.81970861), the Pearl River S&T Nova Program of Guangzhou (No.201710010085), Key Clinical Speciality of Guangzhou Women and Children’s Medical Center, Grant of Institute of Pediatrics of Guangzhou Women and Children’s Medical Center (YIP-2016-022, Pre-NSFC-2018-005).Declaration of Interest: The authors declare that they have no relevant conflicts of interest.Ethical Approval: This study protocols were approved by local ethical committee boards and written informed consent was obtained. All animal care and experimental protocols were approved by local ethics committee boards.

Cellular damage such as oxidation and lipid peroxidation, and DNA damage induced by free-radicals like reactive oxygen species, has been implicated in several diseases. Radicals generated by 2,2-azobis (2-amidino-propane) dihydrochloride (AAPH) are similar to physiologically active ones. In this study we found that polydatin, a resveratrol natural precursor derived from many sources, has the capacity of free radical scavenging and antioxidative damage. Using free radical scavenging assays, the IC50 values of polydatin were 19.25 and 5.29 μg/ml with the DPPH and the ABTS assay, respectively, and 0.125 mg ferrous sulfate/1 mg polydatin with the FRAP assay. With the AAPH-induced oxidative injury cell model assay, polydatin showed a strong protective effect against the human liver tumor HepG2 cell oxidative stress damage. These results indicate that the antioxidant properties of polydatin have great potential for use as an alternative to more toxic synthetic antioxidants as an additive in food, cosmetics and pharmaceutical preparations for the treatment of oxidative diseases.