Plant extracts and microbial-derived molecules, particularly oligo- and polysaccharides, are known to promote tissue repair. Triticum vulgare extract (TVE), derived from germinated whole plants and utilized in medical devices, is rich in bioactive molecules comprised of oligo- and polysaccharides with diverse biological properties beneficial for wound repair. This study investigates the biochemical mechanisms underlying TVE's proregenerative activity in vitro effects, comparing it with hyaluronic acid (HA), a gold standard in wound healing. Preliminary results showed that TVE has scavenging properties/antioxidant activity also at lower concentrations with respect to HA that has long been recognized for these properties (Galvez-Martin et al.). In vitro assays evaluated cell proliferation, migration, extracellular matrix biomarkers, and responses to stress in 2D and 3D models. TVE significantly enhanced keratinocyte migration, reaching approximately 50% wound closure in less than 20 h, whereas the untreated control required about 30 h to achieve the same level of closure. Moreover, TVE performed better than HA at low concentrations, while both treatments improved HDF proliferation and ECM biomarker expression. More specifically, COL1 expression was upregulated by 2.3-fold relative to the control, showing an effect comparable to that induced by HA (2.8-fold). Conversely, ELS expression exhibited a 2.0-fold increase in response to TVE and a 2.6-fold increase following HA treatment compared with the control. In addition, the extract significantly increased HBD-2 expression by 2-fold relative to the control, a novel finding that may be relevant, given the role of this antimicrobial peptide in innate immunity, inflammation regulation, and wound repair. Under stressful conditions, including dehydration and exposure to reactive oxygen species, TVE showed protective effects comparable to HA in both HDF and HaCaT cells. 3D skin models, including those treated with LPS, confirmed the results of 2D assays, further validating TVE's biological properties. Overall, these studies demonstrate that this extract exerts biological effects comparable to pharmaceutical HA in several in vitro skin models. These findings further support the potential application of TVE in wound management and skin regenerative approaches.

Prime editing (PE) enables precise genome modifications without double-strand breaks, yet its application in large mammals has been hindered by low efficiency and the technical bottlenecks of zygote microinjection. This study aimed to establish a scalable PE platform in sheep via the direct electroporation of a PE3 RNP formulation comprising PE2 protein, an epegRNA, and a nicking sgRNA into zygotes. We first validated functional PE2 protein and screened highly efficient enhanced pegRNAs (epegRNAs) targeting two high-value traits, including short-tail (TBXT) and prolificacy (FecB), in primary sheep fibroblasts, achieving editing efficiencies of up to 22.78%. Systematic optimization of zygote electroporation determined that 60 V combined with a PE2: pegRNA mass ratio of 3.5:1 maximized delivery efficiency, yielding gene editing efficiencies of 19.8% and 20.1% for TBXT and FecB, respectively, with no statistically significant differences detected in cleavage or blastocyst rates between electroporated and untreated zygotes. Following dual-target co-delivery, intended edits were detected at both the TBXT and FecB loci in separate preimplantation embryo subsets, with locus-specific editing rates comparable to those observed under single-target conditions. Following the transfer of 54 electroporated blastocysts, 14 live lambs were born. Targeted deep sequencing detected intended edits in five lambs (35.71%), including three at the FecB locus and two at the TBXT locus. Notably, a TBXT-edited lamb exhibiting a distinct short-tail phenotype (15 cm vs. 24.5 cm in wild-type) carried the highest allele frequency, reaching up to 50.97%, providing direct in vivo functional validation. Targeted analysis of predicted off-target sites detected no credible off-target editing, supporting the specificity of this transient RNP delivery strategy. Collectively, this RNP-electroporation framework provides a feasible and precise pathway for scalable prime editing in livestock, bypassing the limitations of conventional injection-based methods.

Surgical intervention and chemotherapy have enhanced the survival rates of patients with advanced gastric cancer. However, the development of chemotherapy resistance remains a major obstacle to effective treatment, leading to poor prognosis in these patients. Increasing evidence suggests that metabolic alterations play crucial roles in mediating chemoresistance. Here we investigate the effects of methylmalonic acid (MMA), a mitochondrial-derived metabolite, on GC progression and oxaliplatin resistance. Analysis of clinical samples revealed that elevated serum MMA levels correlated with chemoresistance and advanced tumor stage in GC patients. Functional assays identified that MMA promoted GC cell proliferation, migration, and resistance to oxaliplatin by attenuating DNA damage. Mechanistically, MMA suppressed oxaliplatin-induced γH2AX expression while upregulating ATAD5 and activating the ATM/ATR-mediated homologous recombination (HR) repair pathway. In vivo studies further confirmed that MMA impaired the antitumor efficacy of oxaliplatin, while silencing PCCA, a key enzyme in MMA synthesis, or supplementing vitamin B12, a regulator of MMA metabolism, reversed MMA-induced chemoresistance. These findings highlight MMA as a potential indicator for oxaliplatin resistance and a therapeutic target in GC. Therefore, targeting MMA through vitamin B12 supplementation could serve as a promising therapeutic strategy to overcome chemoresistance in GC patients with elevated MMA levels.

Piper aduncum L. is a medicinal plant whose essential oil is rich in bioactive constituents, particularly dilapiol, and has attracted interest because of its diverse pharmacological properties. However, its anti-inflammatory, antipyretic, and antinociceptive activities and the mechanisms potentially underlying these effects remain insufficiently characterized. Therefore, this study investigated the pharmacological effects of P. aduncum leaf essential oil (EOPa) and explored potential mechanisms involved in its antinociceptive activity. EOPa was obtained by hydrodistillation and chemically characterized by GC-MS and GC-FID. Its pharmacological effects were evaluated in male Swiss mice using brewer's yeast-induced pyrexia, carrageenan-induced paw edema, acetic acid-induced writhing, and the formalin test. Potential mechanisms of antinociceptive activity were investigated using naloxone, atropine, L-NAME, and glibenclamide as pharmacological antagonists/inhibitors. Chemical characterization identified dilapiol as the major constituent of EOPa (69.65 ± 2.12%). EOPa showed dose-dependent anti-inflammatory, antipyretic, and antinociceptive effects, with the most pronounced activity observed at 200 mg/kg. The essential oil reduced inflammatory edema, fever, and nociceptive responses in the experimental models. EOPa at 200 mg/kg reduced yeast-induced fever by up to 76.1% (p < 0.01), carrageenan-induced paw edema by up to 26.48% (p < 0.0001), and acetic acid-induced writhing by 65.90% (p < 0.001). In the formalin test, EOPa reduced nociceptive behavior by 39.89% during the neurogenic phase and by 70.67% during the inflammatory phase (p < 0.0001). Pretreatment with atropine and glibenclamide markedly attenuated the antinociceptive effect of EOPa, whereas naloxone and L-NAME did not significantly modify the response. These findings provide preclinical evidence that P. aduncum leaf essential oil possesses anti-inflammatory, antipyretic, and antinociceptive activities and suggest that the antinociceptive effect of EOPa involves activation of muscarinic receptors, consistent with a muscarinic agonist-like effect, and stimulation of ATP-sensitive potassium (K+ATP) channels, consistent with a K+ATP-channel-stimulating effect. Further pharmacological, toxicological, and mechanistic studies are warranted to clarify the molecular basis, active constituents, and safety profile of these effects.

Venous thromboembolic disease (VTE), encompassing deep vein thrombosis (DVT) and pulmonary embolism (PE), is associated with substantial morbidity, mortality, and recurrence risk. However, immune-related transcriptomic heterogeneity and recurrence-associated molecular features in VTE remain incompletely defined. This study aimed to explore immune-related transcriptomic clusters associated with VTE recurrence and to prioritize recurrence-associated candidate genes. Public GEO datasets were analyzed, with GSE19151 used as the discovery cohort and GSE48000 used for external validation. Consensus clustering, WGCNA, differential expression analysis, GO/KEGG enrichment, and random forest-based exploratory gene prioritization were performed. The functional relevance of PPP2R1A was further evaluated using a mouse inferior vena cava thrombosis model and endothelial cell experiments, including western blotting, qRT-PCR, CCK-8, Transwell, flow-cytometry, and endothelial-immune co-culture assays. Two exploratory immune-related clusters were identified among VTE patients. Compared with subtype I, subtype II showed stronger IFN-gamma, monocyte, and IL-6 enrichment and a higher recurrence proportion (61.4% vs. 41.7%). Random forest analysis prioritized 23 recurrence-associated candidate genes, including PPP2R1A, although external validation showed modest discriminatory performance (AUC = 0.615). In experimental systems, PPP2R1A expression was associated with thrombus burden, endothelial cell proliferation, migration, apoptosis, inflammatory activation, and altered p-MEK/p-ERK signaling. These findings identify exploratory immune-related clusters in VTE and support PPP2R1A as a recurrence-associated candidate gene linked to endothelial MEK/ERK signaling. Larger prospective cohorts and cell-specific mechanistic studies are needed before these findings can be translated into clinical biomarkers or therapeutic targets.

Myocardial remodeling, an essential pathological development in cardiovascular disease, is largely determined by disruptions in redox homeostasis and mitochondrial bioenergetic function. Iristectorigenin A (ITG), a naturally occurring isoflavone, exhibits a variety of pharmacological effects, including antioxidant and anti-inflammatory activities. While so many protective effects, its specific effect on cardiac remodeling has not been reported previously. In this study, we aims to investigate the role of ITG in pressure overload-induced cardiac remodeling with a particular focus on mitochondrial function and oxidative stress. We revealed that ITG mitigated TAC-induced cardiac remodeling and improved cardiac function, with the 20 mg/kg dose showing optimal efficacy; further escalation to 40 mg/kg provided no additional benefit. ITG attenuated cardiomyocyte hypertrophy and inhibited activation of fibroblasts in vitro. Additionally, ITG treatment effectively suppressed oxidative stress and inflammation in the hearts of TAC mice, while improving mitochondrial morphology and the integrity of inner membrane structure. Notably, surface plasmon resonance (SPR) analysis indicated a direct interaction between SIRT6 and ITG with micromolar affinity. Besides, sustained TAC- or PE-stimulation impaired cardiac sirtuin 6 (SIRT6) protein levels, which were partially reversed by ITG. More importantly, silencing SIRT6 and SIRT6 knockdown separately inhibited the protective effects of ITG in vitro and in vivo. Mechanistically, ITG promotes SIRT6 levels by inhibiting its ubiquitination degradation. Furthermore, the cardioprotective effect of ITG was concomitant with SIRT6 upregulation, deacetylation and dephosphorylation of forkhead box protein O1 (FOXO1) and the subsequent increased transcription of antioxidant genes. Collectively, we firstly discover that ITG inhibits oxidative stress and safeguards mitochondrial morphofunctional homeostasis in response to pathological stress through activating SIRT6/FOXO1 axis, suggesting the therapeutic potential of ITG in the treatment of cardiac hypertrophy and remodeling, providing a promising avenue for future research and potential clinical application. Schematic illustration of ITG's potential protective mechanism against cardiac hypertrophy.

Lung cancer surgery has evolved substantially, but safety alone does not define quality. Using the Avedis Donabedian model, quality encompasses structure, process, and outcomes. Structural factors include surgeon specialization, institutional volume, and multidisciplinary care. Process indicators involve guideline-concordant staging, systematic lymphadenectomy, and complete resection; inadequate nodal assessment or uncertain resection negatively affects survival. Outcomes extend beyond mortality to oncologic results and patient-centered measures. Composite metrics such as textbook outcome and failure-to-rescue support benchmarking, while society-driven databases enable risk-adjusted comparisons. Current frameworks emphasize perioperative and technical metrics, but underrepresent patient-reported outcomes, long-term survival, and multimodality care.