Colorectal cancer (CRC) liver metastasis (LM) is a leading cause of cancer-related death, with pre-metastatic niche (PMN) formation being critical for metastasis. Insulin-like growth factor binding protein 2 (IGFBP2) is overexpressed in metastatic CRC, but its role in intrahepatic fibrotic PMN formation and CRC-LM remains unclear. Bioinformatics analyses and tissue microarray samples were conducted to assess the expression level and prognostic significance of IGFBP2 in CRC patients. After overexpression of IGFBP2, its in vivo metastatic-promoting ability was evaluated by establishing a splenic injection liver metastasis model, while its in vitro malignant properties were assessed via colony formation, proliferation assay, flow cytometry, wound-healing assay and transwell assay. Furthermore, flow cytometry, RT-qPCR and ELISA were used to evaluate the effect of IGFBP2 overexpression on the polarization of intrahepatic macrophages. In addition, Western blot and immunofluorescence were performed to detect the effect of M2-polarized macrophages on hepatic stellate cells (HSCs). Finally, the splenic injection liver metastasis model was treated with CWI1-2 (IGFBP2 inhibitor) or pirfenidone (PFD, a TGF-β signaling modulator), and the therapeutic effects were observed. We found that IGFBP2 is overexpressed in CRC tissues, correlating with poor prognosis. Meanwhile, IGFBP2 overexpression significantly promotes CRC-LM progression in vivo. Mechanistically, it drives M2 polarization of intrahepatic macrophages to boost TGF-β secretion. Subsequently, TGF-β activates HSCs, elevates the deposition of extracellular matrix (α-SMA, COL1, FN), and ultimately contributes to the development of intrahepatic fibrotic PMN. Notably, CWI-2 or PFD could suppress CRC-LM, with combined treatment showing synergistic effects both in vitro and in vivo. IGFBP2 is highly expressed in CRC and correlated with patient prognosis. CRC-derived IGFBP2 drives intrahepatic fibrotic PMN formation via intrahepatic macrophages M2 polarization and TGF-β-mediated HSCs activation, which in turn facilitates CRC-LM. Furthermore, selective targeting of the process of intrahepatic PMN formation holds promise as a viable strategy for the prevention and treatment of CRC-LM.

Osteoarthritis (OA) is a prevalent age-related disease associated with significant pain and disability. Although 5-methylcytosine (m5C) modification is implicated in age-related diseases, its role in OA remains unclear. Here, the regulatory effects and mechanisms of the m5C-related proteins NSUN2 and ALYREF on chondrocyte senescence and inflammation during OA progression were investigated. NSUN2 expression and m5C modification were upregulated in articular chondrocytes from damaged cartilage of surgically induced OA and naturally aged mice, as well as in primary human chondrocyte aging models. NSUN2 was found to promote chondrocyte senescence and NLRP3 inflammasome activation. Messenger RNA (mRNA) m5C enrichment, RIP sequencing, and transcriptomic sequencing elucidated the mechanism by which NSUN2 regulates inflammation and aging. IP3R3 was identified as a target gene of NSUN2. Rescue experiments showed that NSUN2 induced Ca2+ overload, which was mitigated by 2-aminoethoxydiphenyl borate (2-APB) or BAPTA/AM. NSUN2 knockdown or IP3R3 inhibition protected mouse articular cartilage from senescence and NLRP3 inflammasome activation, alleviating OA progression. Mechanistically, NSUN2 cooperated with the m5C reader ALYREF to stabilize and promote cytoplasmic export of IP3R3 mRNA, increasing IP3R3 expression. Thus, NSUN2 inhibition reduces chondrocyte senescence and cartilage damage via the IP3R3-Ca2+ axis, may represent a potential therapeutic target for further investigation.

BackgroundInitially recognized for its role in hematopoiesis, Interleukin-11 (IL-11) is now understood to possess a wide spectrum of biological activities. It is involved in critical physiological processes, including immune cell differentiation, inflammatory response regulation, and tissue repair. Consequently, IL-11 is strongly implicated in the pathogenesis and progression of various chronic diseases.DiscussionThis review examines the multifaceted biology of IL-11, detailing its discovery, molecular structure, and signaling mechanisms. We also explore its expression patterns across different tissues. A primary focus is placed on elucidating the critical role of IL-11 in the pathogenesis and progression of various chronic diseases. Furthermore, we discuss the emerging therapeutic potential of targeting the IL-11 pathway, evaluating evidence from both experimental models and clinical studies.ConclusionBy synthesizing current knowledge on the biological characteristics and disease associations of IL-11, this review aims to provide a comprehensive theoretical foundation for future research into its role in chronic diseases and its potential as a therapeutic target.

Hepatocellular carcinoma (HCC) remains a major global health burden with limited therapeutic options and poor prognosis. PDRG1 is upregulated in several malignancies, yet its clinical relevance and mechanistic role in HCC are not fully understood. Here, we investigated the contribution of PDRG1 to HCC progression and delineated the underlying molecular mechanism. Using public datasets, patient specimens, in vitro functional assays, and subcutaneous xenograft models, we evaluated PDRG1 expression, biological functions, and downstream pathways. Transcriptome profiling, pathway enrichment analysis, rescue experiments, co-immunoprecipitation, and ChIP-qPCR were performed to define the PDRG1-EZH2-p21 axis. PDRG1 was significantly upregulated in HCC tumor tissues compared with adjacent non-tumor liver tissues and was associated with worse patient survival. Functionally, PDRG1 enhanced HCC cell proliferation, migration, invasion, colony formation, and tumor growth in vivo. RNA-seq and enrichment analyses identified cellular senescence as a prominent downstream program regulated by PDRG1. Mechanistically, PDRG1 directly interacted with EZH2, increased H3K27me3 enrichment at the p21 promoter, and suppressed p21 transcription. Restoration of p21 expression attenuated the oncogenic effects of PDRG1, whereas EZH2 overexpression rescued the impaired malignant phenotypes caused by PDRG1 knockdown. Domain-mapping further indicated that the N-terminal residues 36-70 of PDRG1 contribute to its interaction with EZH2. Collectively, our findings identify PDRG1 as a clinically relevant oncogene in HCC and reveal an epigenetic mechanism by which PDRG1 cooperates with EZH2 to repress p21 and bypass senescence. The PDRG1-EZH2-p21 axis may represent a potential biomarker and therapeutic target for HCC.

Diabetic foot ulcers (DFUs) are a major complication associated with diabetes and frequently result in serious health consequences. The cGAS-STING pathway has been linked to various disorders through its modulation of inflammation and ferroptosis, yet its specific function in the context of DFUs remains poorly understood. This work sought to explore the impact of targeting cGAS to inhibit STING activation on DFU healing, as well as its regulatory role in angiogenesis and endothelial cell ferroptosis. We observed elevated levels of cGAS and STING in wound samples from DFU subjects and animal models. Knockdown of cGAS significantly accelerated wound healing in DFU rat models, improved histopathological changes, increased collagen synthesis, and enhanced angiogenesis. Furthermore, DFU tissues in rats with cGAS knockdown exhibited notably lower levels of STING, p-TBK1, p-IRF3, p-P65, and pro-inflammatory mediators. In high glucose (HG)-stimulated vascular endothelial cells, cGAS and STING expression was upregulated. Silencing cGAS effectively inhibited HG-induced STING activation, while enhancing cell viability, migration, and tube formation. Furthermore, HG-induced ferroptosis in vascular endothelial cells, characterized by increased lipid peroxidation and altered ferroptosis-related protein expression, was significantly reversed by cGAS knockdown, indicating its role in inhibiting ferroptosis. Notably, the addition of a ferroptosis inducer in cGAS-knockdown cells partially reversed the improvement in angiogenesis. In conclusion, cGAS knockdown effectively accelerates DFU wound healing by inhibiting STING activation and modulating ferroptosis in endothelial cells, underscoring the pivotal role of the cGAS-STING-ferroptosis axis in DFU progression and healing, with potential therapeutic implications.

Kidney transplantation remains the gold standard for end-stage renal disease, but ischemia-reperfusion injury and delayed graft function continue to hinder outcomes. Growth differentiation factor 15 (GDF15), a stress-responsive cytokine from the transforming growth factor-β superfamily, is upregulated in response to cellular injury and hypoxia. Although GDF15 has been studied in acute kidney injury and sepsis, its role in kidney transplantation remains unclear. In this study, we combined transcriptomic analysis of human kidney allografts with murine models to investigate GDF15's role in transplant injury. GDF15 was upregulated in renal tubular epithelial cells, particularly in delayed graft function, and its levels in urine correlated with serum creatinine levels, indicating a link to graft dysfunction. In syngeneic and allogeneic murine transplant models, GDF15 deficiency worsened tubular injury and inflammation, while recombinant GDF15 protected against injury and promoted an anti-inflammatory M2 macrophage phenotype. We also identified activating transcription factor 4 as a key regulator of GDF15 in renal stress, with its knockout reducing GDF15 expression and worsening transplant injury. Macrophage depletion confirmed that macrophage-mediated inflammation was a major factor in GDF15-deficient graft injury. In conclusion, GDF15 regulates kidney transplant injury by modulating macrophage polarization, making it a potential therapeutic target for improving transplant outcomes.

BACKGROUND:Myocardial ischemia-reperfusion (MIR) injury compromises therapeutic effects of revascularization and leads to functional impairment and exacerbation of structural damage in the heart. Limiting the damage caused by MIR is crucial but is still an unmet clinical need because of the complexity of the underlying mechanisms. Increasing evidence suggests that lysosomal autophagy plays a significant regulatory role in MIR injury. The specific mechanisms involved remain to be fully understood. METHODS:We here systematically analyzed the murine MIR model database to screen the potentially protective lysosome-localized proteins against MIR injury. The positive hits were further functionally screened and validated for their capability on autophagy and hypoxia/reoxygenation insults of cardiomyocytes. After exploring the detailed molecular mechanism underlying the protective effects of the target protein, we generated target gene cardiac-specific knockout mice and overexpression mice to verify its function in mouse MIR injury models. RESULTS:LAPTM4A (lysosome-associated protein transmembrane 4 alpha) stood out as a significant protective lysosome-localized protein from the screening. LAPTM4A deficiency significantly heightened the inflammatory response and cell death both in primary cardiomyocytes and in a MIR-induced mouse model. Conversely, LAPTM4A overexpression exerted protective effects on cell viability and myocardial damage. Mechanistically, LAPTM4A interacts with Rubicon (Run domain Beclin1-interacting and cysteine-rich domain-containing protein), hindering its engagement within the Beclin1 complex, resulting in a robust augmentation of autophagic flux and thereby mitigating cardiac damage during reperfusion. It is important to note that Rubicon knockdown markedly reversed the aggravated injury induced by LAPTM4A knockdown, further verifying the effects of LAPTM4A depend on Rubicon. CONCLUSIONS:Our findings screened out and validated that LAPTM4A is a lysosome-localized protein exerting protective effects against MIR injury by facilitating autophagic flux. Targeting LAPTM4A represents a promising therapeutic strategy for mitigating MIR injury.

Rationale:During ischemia‒reperfusion injury (IRI), BMAL1 has been shown to alleviate inflammation and kidney damage. However, the function of the tubular epithelium-macrophage interaction mediated by BMAL1 in IRI-induced renal fibrosis is still unclear. Methods:A mouse model of kidney-specific BMAL1 overexpression was developed to study how BMAL1 affects renal fibrosis, exosome production, and the macrophage-to-myofibroblast transition (MMT). The role of exosomes in the MMT and renal fibrosis was examined in both in vitro and in vivo studies using exosomes extracted from TCMK-1 cells. Exosomes from BMAL1-overexpressing TCMK-1 cells subjected to hypoxia-reoxygenation (H/R) were isolated and subjected to miRNA sequencing to identify key exosomal components. Exosomal miR-27a-3p regulation by BMAL1 and its downstream effects on TGFBR1/smad3 in macrophages were investigated using a variety of experimental methods. To assess the effect of exosomal miR-27a-3p on MMT and renal fibrosis, additional in vitro and in vivo investigations were conducted. Results:Renal IRI increased exosome secretion, promoted MMT, and exacerbated renal fibrosis, whereas BMAL1 overexpression or Rab27a knockout significantly attenuated IRI-induced MMT and fibrotic progression. Exosomes derived from H/R-treated tubular epithelial cells further exacerbated MMT and renal fibrosis in an IRI model. Notably, tubular-specific overexpression of BMAL1, elevation of exosomal miR-27a-3p levels, or inhibition of exosome secretion significantly attenuated the progression of both MMT and fibrosis. Mechanistic studies demonstrated that BMAL1 binds directly to the miR-27a-3p promoter region, enhancing transcription. Exosomal miR-27a-3p subsequently targets TGFBR1 mRNA in macrophages, thereby suppressing the TGFBR1/smad3 signaling pathway and ultimately attenuating MMT and renal fibrosis. Conclusions:BMAL1 expression was suppressed in IRI, which promoted MMT and renal fibrosis via the exosomal miR-27a-3p-TGFBR1/smad3 pathway. Targeting this signaling pathway may offer a potential therapeutic strategy for alleviating IRI-induced renal fibrosis.

Background: Osteoarthritis (OA) is a prevalent degenerative joint disease marked by the continuous deterioration of cartilage and functional limitations, impacting millions, with few disease-modifying treatments available. The extracellular matrix protein hyaluronan and proteoglycan link protein 1 (HAPLN1) stabilises aggrecan-hyaluronan complexes in cartilage; nonetheless, its function in osteoarthritis pathogenesis is inadequately comprehended. This research examined HAPLN1 expression in osteoarthritis models and its impact on chondrocyte senescence. Methods: Primary articular mouse chondrocytes were extracted from neonatal C57BL/6 mice and grown in DMEM/F12 conditions. OA-like conditions were produced by TNF-α (10 ng/ml) or tert-butyl hydroperoxide (TBHP, 50 μM). HAPLN1 knockdown was accomplished using lentiviral shRNA transfection. Recombinant mouse HAPLN1 (rmHAPLN1, 2.5 ng/ml) was employed for rescue. SA-β-Gal staining, p16/p21 expression, and telomerase activity were used to measure senescence. Matrix breakdown indicators (MMP-13, Col2a1) and inflammatory cytokines (CXCL10, IL-1β, MCP-1) were evaluated by qPCR, Western blot, and ELISA. Oxidative stress was measured by ROS levels, γ-H2AX, SOD/CAT activity, and Foxo3a phosphorylation/localization. In vivo , post-traumatic OA was generated by destabilisation of the medial meniscus (DMM) in 8-week-old male mice, with intra-articular lentiviral HAPLN1 administration at weeks 1 and 4. Krenn’s approach was used to assess synovitis, whereas OARSI grading was used to score cartilage integrity. Results: TNF-α and TBHP-stimulated chondrocytes and cartilage from DMM-operated animals showed significantly lower levels of HAPLN1 mRNA and protein. Silencing HAPLN1 dramatically accelerated extracellular matrix breakdown, inflammatory cytokine release, cellular senescence, and oxidative stress, which was mediated by increased Foxo3a phosphorylation at Ser253/Thr32 and cytoplasmic retention. In contrast, supplementation with rmHAPLN1 or intra-articular lentiviral-mediated HAPLN1 overexpression improved these changes both in vitro and in vivo , considerably decreasing OARSI cartilage damage and synovitis scores when compared to untreated OA controls. Conclusion: Our results show that HAPLN1 protects against OA by reducing Foxo3a-induced chondrocyte senescence and oxidative stress, making it a suitable therapeutic target.

BACKGROUND:Sepsis, driven by dysregulated host inflammation, remains a leading cause of global mortality and lacks sufficiently effective therapies. Phillyrin (PHN), a lignan glycoside from Forsythia suspensa (Thunb.) Vahl (Oleaceae), exhibits anti-inflammatory and antimicrobial properties. However, its molecular mechanism in sepsis remains poorly understood. OBJECTIVE:To delineate the molecular pathways by which PHN mitigates sepsis. METHODS:Reverse transcription-quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, cytotoxicity assessments, and calcein acetoxymethyl ester/propidium iodide staining were employed to evaluate the protective effects of PHN against lipopolysaccharide (LPS)-mediated pyroptosis in human pulmonary microvascular endothelial cells (HPMECs). Network pharmacology and integrative bioinformatics were used to identify candidate regulatory axes. Findings were then validated across multiple models: HPMEC-differentiated human promyelocytic acute leukemia cell co-cultures systems, Tg(mpx:GFP) zebrafish, and BALB/c mice. Validation techniques comprised luciferase reporters, pharmacological modulators, and morpholino knockdown. RESULTS:PHN significantly attenuated LPS-induced inflammatory pyroptosis in HPMECs. Evidence included reduced proinflammatory cytokine production, decreased lactate dehydrogenase leakage, and fewer pyroptotic cells. Computational analyses identified the microRNA (miR)-203a-caspase-4 (CASP4) axis as a primary mediator of PHN's anti-septic activity. In co-culture systems, PHN suppressed cytoplasmic LPS-triggered pyroptosis through miR-203a-dependent CASP4 downregulation, thereby reducing inflammatory cytokine secretion and neutrophil recruitment. In LPS-challenged zebrafish, PHN upregulated miR-203a to suppress caspase-B (CASPB), diminishing cytokine expression and neutrophil migration while improving survival. In murine sepsis models induced by LPS or cecal ligation and puncture, PHN modulated the miR-203a-caspase-11 (CASP11) axis to confer multiple therapeutic benefits. These included improved survival rates, stabilized body temperature, reduced bronchoalveolar lavage protein levels and neutrophil infiltration, attenuated multi-organ injury, and decreased systemic cytokine levels. CONCLUSION:These data identify the miR-203a-CASP4/11/B axis as a critical mediator of endothelial pyroptosis in sepsis. PHN attenuates sepsis by upregulating miR-203a to inhibit CASP4/11/B-dependent pyroptosis. Therefore, PHN warrants further investigation as a potential therapeutic agent for sepsis.

Cervical cancer is a common gynecological malignancy, with a 5-year survival rate of only 17% for recurrent or metastatic cases. Increased extracellular matrix stiffness, a key change in the tumor mechanical microenvironment, promotes tumor metastasis via mechanotransduction. Piezo1, a mechanosensitive cation channel, senses matrix stiffness and converts mechanical signals into intracellular chemical signals. Neutrophil extracellular traps (NETs) are overformed in tumors, but the mechanism by which matrix stiffness regulates NETs in cervical cancer remains unclear. We detected matrix stiffness and related protein expression in cervical cancer tissues using atomic force microscopy and histochemical staining. Polyacrylamide gel models were used to culture HeLa/SiHa cells, with transcriptome sequencing and ELISA to analyze IL-8 expression. NETs were induced from human peripheral blood neutrophils, and their effect on lymphatic endothelial cells was evaluated. A TC-1 mouse model was used to verify in vivo effects, and Western blot/ELISA explored the Piezo1/NF-κB pathway. Higher Young’s modulus, increased α-SMA/Collagen I expression and collagen content in metastatic cervical cancer tissues. High matrix stiffness activated Piezo1/NF-κB, upregulated IL-8, induced NETs, and enhanced lymphatic endothelial cell tube formation/migration. BAPN reduced tumor stiffness, inhibited metastasis, and decreased NETs in mice. Knocking down Piezo1 blocked NF-κB activation and IL-8 upregulation. High matrix stiffness activates Piezo1/NF-κB to promote IL-8 secretion and NETs formation, enhancing lymphangiogenesis and cervical cancer metastasis, providing a new target for advanced cervical cancer treatment.

Microglia-mediated neuroinflammation plays a key role in the process of ischemic stroke. Lymphocyte adaptor protein (LNK) negatively regulates cytokine signaling in peripheral immune cells, but its roles in neuroinflammation and ischemic stroke remain unknown. Here, we demonstrated that LNK expression was decreased in peripheral blood mononuclear cells and microglia after stroke in humans and mice, respectively. Microglia-specific knockout of LNK in mice promoted neuroinflammation and exacerbated ischemia‒reperfusion (I/R) injury in a mouse model of ischemic stroke. Mechanistically, LNK and pyruvate kinase M2 (PKM2) can competitively bind to the OTU deubiquitinase, ubiquitin aldehyde binding 1 (OTUB1), and promote the ubiquitination and degradation of PKM2. In the case of LNK deficiency, OTUB1 can bind to PKM2 and increase its stability, which promotes microglial glycolysis and lactate secretion, enhances the expression of H4K12la and proinflammatory cytokines, and ultimately, worsens brain I/R injury. On the basis of these findings, a PKM2 inhibitor was shown to reduce brain I/R injury exacerbated by LNK deficiency. Moreover, adenovirus-mediated LNK overexpression significantly protected against brain I/R injury in a mouse model of ischemic stroke. Therefore, our study reveals a new function of LNK, which acts as a competitive inhibitory molecule to exert negative regulation, thereby determining stroke outcomes.

Enveloped virus invasion relies on spike glycoprotein-mediated membrane fusion. Cholesterol that serves crucial roles in modulating protein conformations and membrane properties, plays an essential role in the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cell entry. However, the precise regulatory mechanism of cholesterol in SARS-CoV-2 fusion remains unknown. Here, using an in vitro vesicle-vesicle content mixing assay, we demonstrated that the addition of cholesterol enhanced SARS-CoV-2 spike-mediated vesicle-vesicle fusion, with this enhancement being dependent on the C-terminal cytoplasmic domain of spike. Further single-vesicle analyses demonstrate this enhancement primarily stems from increased docking probability, with cholesterol exerting mild effect on fusion probabilities. In the cell-based membrane fusion assay, cholesterol depletion from spike containing membrane significantly reduces syncytia formation and SARS-CoV-2 pseudovirus infection, indicating its modulatory role in this process. Using structured illumination microscopy (SIM) based super-resolution imaging and single-molecule photobleaching microscopy, we demonstrated that spike proteins tended to form into an oligomeric cluster in the presence of cholesterol, likely through the interaction between cholesterol and palmitoylated cysteine rich region (CRR) in the C-terminus of spike. Last, substitution of residues of CRR with alanine in the C-terminus of spike abolished both the cholesterol-induced spike clustering and the cholesterol-dependent enhancement of vesicle docking. Taken together, our results suggest that cholesterol may induce the oligomerization of spike through specific interactions with its CRR, with this structural clustering critically mediating viral docking to host cell membranes, thereby promoting the subsequent membrane fusion and viral entry processes.

Ovulatory dysfunction is a typical symptom of polycystic ovary syndrome (PCOS) and is widely recognized as a major obstacle to fertility. Numerous studies have reported a close association between the programmed cell death (PCD) of granulosa cells (GCs) and ovulatory dysfunction in PCOS. Recently, a novel type of PCD exhibiting features of pyroptosis, apoptosis, and necroptosis was identified and termed PANoptosis. However, whether the PANoptosis occurs in PCOS GCs and plays a role in ovulatory dysfunction remains unclear. In this study, we found that the expression levels of PANoptosis-related molecules (NLRP3, CASP1, IL-1β, CASP3, MLKL), the PANoptosis regulator AIM2, IFN-γ, the IFN-responsive transcription factor 1 (IRF1), and HAS2 were significantly increased in GCs collected from PCOS patients, DHT-treated KGN cells, and the ovaries of DHEA-induced PCOS mouse models. Most importantly, KGN cells treated with a combination IFN-γ and lipopolysaccharide (LPS) exhibited similar effects. While, blocking IRF1 (using RNA interference-based knockdown), HAS2 (using 4-MU), JAK2 (using AZD-1480), or STAT3 (using SH-4-54) completely abolished these effects. Mechanistically, we revealed that elevated IFN-γ promotes AIM2 expression and AIM2-PANoptosome assembly through the JAK2/STAT3/IRF1/HAS2 axis. Further in vivo studies showed that inhibiting HAS2 (using 4-MU) and AIM2 (using A151) increased the number of oocytes retrieved from DHEA-induced PCOS mice. Taken together, our results suggest that AIM2-PANoptosis occurs in the GCs of PCOS and is triggered by the IFN-γ/IRF1/HAS2 signalling pathway, thereby resulting in ovulatory dysfunction. This study therefore provides valuable potential therapeutic targets for the treatment of this typical PCOS symptom.

A major obstacle in knocking down oncogenes for tumor therapy is the efficient delivery of siRNA into the cytosolic spaces of cancer cells. Here, we genetically bioengineer biomimetic nanovesicles with tumor-recognition and enzyme-controlled membrane fusion functions for efficiently delivering small interfering RNA into cancer cells towards gene silencing tumor therapy. The siRNA@eS-BNVs are formulated by encapsulating siRNA inside the core and coating with genetically engineered HEK293TACE2- cell membranes encoded with functional S protein, which can recognize cancer cells and initiate membrane fusion when triggered by the enzyme. The siRNA@eS-BNVs demonstrate better efficacy for cytosolic siRNA delivery and RNA interference than conventional formulations. By intravenous injection, siRNA@eS-BNVs are highly accumulated in tumors and potently inhibited tumor and lung metastasis by simultaneously silencing the epidermal growth factor receptor gene in vivo. The cancer cell-targeting and enzyme-activatable nanovesicles provide a valuable strategy for effective and precise drug delivery.

AimThis study aims to investigate the benefits of gypensapogenin I (GI) on myocardial ischemia–reperfusion injury (MIRI) and the underlying mechanisms.MethodsAn MIRI model was established by ligating the anterior descending coronary artery (LAD) followed by blood flow restoration in mice. Cardiac dysfunction and myocardial infarction size were evaluated by echocardiography and triphenyltetrazolium chloride (TTC) staining. PANoptosis, ferroptosis, and mitochondrial redox state were examined by immunofluorescence, Western blotting, and an ELISA kit. In addition, molecular and biochemical methods were applied to illustrate the exact mechanisms of GI on MIRI.ResultsGI pretreatment alleviated cellular oxidative stress, inhibited PANoptosis and ferroptosis, reduced myocardial infarction area, and improved cardiac function during MIRI. Further results revealed that mitochondrial biogenesis and the anti-oxidative system were impaired in mice suffering from MIRI, and these effects were significantly alleviated by GI treatment via downregulation of the NADPH oxidase 2 (NOX2) level. Moreover, NOX2 promoted mitochondrial dysfunction by suppressing the AMP-activated protein kinase (AMPK)–PGC-1α–Sirt3 signaling pathway. In addition, the NOX2 activator exacerbated oxidative damage and offset all the beneficial effects of GI on mitochondrial function, PANoptosis, and ferroptosis. Meanwhile, reinforced AMPK phosphorylation by GI or AMPK activator (5-aminoimidazole-4-carboxamide ribonucleotide, AICAR) maintained the mitochondrial redox state and biogenesis and suppressed PANoptosis and ferroptosis.ConclusionGI pretreatment protected the cardiomyocytes from MIRI-induced PANoptosis and ferroptosis by maintaining the mitochondrial redox state and biogenesis through the modulation of the NOX2/AMPK signaling pathway. Our findings indicate that GI pretreatment could be a promising therapeutic agent for MIRI treatment.

Background:There is limited research about the pregnancy outcomes of POSEIDON group 2 patients. This study aims to evaluate the factors related to the clinical outcomes of in vitro fertilization (IVF) and/or intracytoplasmic sperm injection (ICSI) in POSEIDON group 2 patients, so as to provide clinical guidance to improve clinical outcomes. Methods:A total of 1,249 IVF/ICSI cycles of POSEIDON group 2 patients were retrospectively analyzed from July 2019 to September 2024. The patients were divided into different groups according to live birth, maternal age, and endometrial thickness (EMT). The clinical outcomes were compared between groups. Results:Compared with the non-live birth group, maternal age, gonadotropin (Gn) dosage, and Gn total dosage were significantly lower, while EMT was significantly higher in the live birth group (p < 0.05). The type of infertility showed statistically significant differences (p = 0.013). Multivariate logistic regression analysis indicated that maternal age (OR = 1.195, 95% CI: 1.077-1.327; p = 0.001) and EMT on transfer day (OR = 0.887, 95% CI: 0.820-0.959; p = 0.003) were independent influencing factors for live birth in POSEIDON group 2 patients. According to the receiver operating characteristic (ROC) curve predictive cutoff value, the patients were divided into group A (age < 38 years, n = 249) and group B (age ≥ 38 years, n = 155). Group A had a higher human chorionic gonadotropin (hCG)-positive rate (55.8% vs. 34.2%, p < 0.001), embryo implantation rate (35.2% vs. 19.2%, p < 0.001), clinical pregnancy rate (47.8% vs. 29.7%, p < 0.001), and live birth rate (37.3% vs. 20.0%, p < 0.001) and a lower early miscarriage rate (12.6% vs. 28.3%, p = 0.016) than group B. According to the ROC predictive cutoff value of EMT, group A patients were further divided into group A1 (EMT < 11.65 mm, n = 136) and group A2 (EMT ≥ 11.65 mm, n = 113). Group B patients were divided into group B1 (EMT < 11.65 mm, n = 93) and group B2 (EMT ≥ 11.65 mm, n = 62). Compared with group A1 and group A2, group B1 had a lower hCG-positive rate (28.0% vs. 49.3%, p = 0.001; 28.0% vs. 63.7%, p < 0.001), embryo implantation rate (14.7% vs. 32.2%, p < 0.001; 14.7% vs. 40.2%, p < 0.001), clinical pregnancy rate (23.7% vs. 41.2%, p = 0.006; 23.7% vs. 55.8%, p < 0.001), and live birth rate (16.1% vs. 33.8%, p = 0.003; 16.1% vs. 41.6%, p < 0.001). Conclusions:Maternal age and EMT on transfer day are independent factors affecting IVF/ICSI clinical outcomes in POSEIDON group 2 patients. Among these, maternal age <38 years or EMT ≥ 11.65 mm indicated better clinical outcomes. Further stratification by age and EMT on transfer day may optimize clinical outcomes in POSEIDON group 2 patients.

Renal ischemia–reperfusion injury triggers substantial inflammatory reactions, with renal tubular epithelial cells (TECs) and macrophages playing crucial roles. Extracellular vesicles (EVs) are key mediators of intercellular signaling. However, the precise roles and mechanisms by which TEC-derived EVs influence macrophage functions remain unclear. This study investigated how miR-491-3p within EVs from TECs modulates macrophage polarization, thus worsening renal inflammation and damage. In models that are in vitro and in vivo, EVs enriched with miR-491-3p from TECs subjected to hypoxia/reoxygenation were shown to promote M1 polarization in macrophages, enhancing inflammatory responses and inducing TEC apoptosis. Mechanistically, miR-491-3p directly targets sirtuin 1 (SIRT1) in macrophages, inhibiting SIRT1-mediated Notch intracellular domain (NICD) deacetylation, thereby regulating NICD stability. This regulation blocks F-box and WD repeat domain-containing 7 (FBXW7)–NICD binding, reduces NICD ubiquitination, and activates the Notch/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway. Furthermore, Rab27a knockout, which limits EVs’ release, substantially reduces M1 macrophage polarization and renal tissue damage. These findings indicate that miR-491-3p from TEC-derived EVs targets SIRT1, inhibiting the deacetylation of NICD mediated by SIRT1, which subsequently prevents ubiquitin-mediated NICD degradation. This mechanism modulates the inflammatory phenotype of macrophages and promotes the inflammatory response, thereby worsening renal injury induced by ischemia–reperfusion injury and highlighting a potential therapeutic target.

Acute lung injury(ALI)is characterized by a sudden decline in pulmonary gas exchange function due to various pathological fac-tors,with severe cases progressing to acute respiratory distress syndrome(ARDS).ARDS affects approximately 3 million patients annually,accounting for 10％of intensive care unit admissions[1].No specific drugs have been developed for this condition yet.