Traumatic brain injury (TBI) is a major cause of morbidity, mortality, and long-term neurological impairment. Growing evidence suggests that the gut-brain axis plays a crucial role in TBI pathophysiology, with gut microbiota dysbiosis potentially inducing inflammation and aggravating brain injury. Lycopene (Lyc), a potent antioxidant derived from tomatoes with neuroprotective and anti-inflammatory properties, is considered a promising potential therapy for TBI. Sunflower oil (SO) can effectively enhance Lyc bioavailability. We established a TBI mouse model and treated it with SO-loaded Lyc (SO-Lyc). Serum and fecal samples were collected for metabolomic analysis, 16S rRNA sequencing for gut microbiota changes, ELISA for inflammatory cytokines and injury markers, H&E staining for gut/brain histopathology, and immunohistochemistry for intestinal barrier-related gene expression. Furthermore, we used 16S rRNA data from patient samples with TBI to perform association analyses. The results showed that SO-Lyc supplementation may mitigate damage to intestinal and cerebral tissues by restoring intestinal barrier function and suppressing systemic inflammation through modulation of key metabolic pathways, such as those involving linoleic acid, sphingolipids, and tryptophan. Its effects manifest as reduced serum inflammatory cytokines (TNF-α, IL-6, CRP, IL-1β), intestinal injury markers (DAO, D-LA, I-FABP, ET), and brain injury markers (NFL, GFAP, S100B, NSE, UCH-L1), while enhancing the expression of intestinal barrier genes (occludin and ZO-1). Furthermore, this supplement promotes the growth of gut microbiota with anti-inflammatory properties. Differences in inflammation and gut microbiota have been observed among TBI patients with varying degrees of severity, suggesting that Lyc containing SO may be an effective treatment for TBI patients.

Skin fibrosis is a chronic inflammatory disorder characterized by excessive extracellular matrix (ECM) deposition and aberrant fibroblast activation, leading to tissue stiffening and progressive functional impairment. Despite its clinical prevalence, no FDA-approved therapies are currently available, highlighting a significant unmet medical need. The present study investigated the anti-fibrotic potential of yohimbine using an integrated network pharmacology approach and a bleomycin (BLM)-induced murine skin fibrosis model. Network pharmacology identified 168 overlapping targets between yohimbine and skin fibrosis, while KEGG enrichment highlighted the PI3K/Akt and MAPK pathways as key antifibrotic signaling mechanisms. Subcutaneous administration of BLM (3 mg/kg) for 28 days induced marked skin fibrosis compared with sham control. Yohimbine treatment at 2.5 and 5 mg/kg attenuated bleomycin (BLM)-induced skin fibrosis, with the 5 mg/kg dose demonstrating a more consistent antifibrotic response compared with the BLM group. Molecular analyses demonstrated that yohimbine suppressed the pro-fibrotic (Ccn2, α-Sma, Vimentin, Zeb, and Snail) and inflammatory markers (Tnf-α) expression. Furthermore, yohimbine restored the expression of antifibrotic (Hgf) and antioxidant (Gstp1, Nrf2, and Hmox1) markers, while reducing ECM-associated markers, including Fibronectin (FN1) and Collagen Type III (COL3α1), at both mRNA and protein levels. Histopathological, Masson's trichrome, and Sirius red/fast green collagen staining analysis further demonstrated that yohimbine treatment significantly (p < 0.05) reduced collagen deposition and restored normal skin architecture. Collectively, these findings demonstrate that yohimbine attenuates skin fibrosis by suppressing inflammatory and fibrotic responses, potentially through modulation of PI3K/Akt-MAPK signaling, and may represent a potential lead for further investigation in fibrotic skin disorders.

The ubiquitin-specific peptidase 42 (USP42) is a cysteine-dependent deubiquitinase with pro-tumorigenic roles in several cancers. In lung adenocarcinoma (LUAD), elevated USP42 expression correlates with advanced stage and poor survival, mirroring the prognostic impact of prostaglandin-endoperoxide synthase 2 (PTGS2) overexpression. However, the substrate repertoire and mechanisms by which USP42 drives malignant progression and immune evasion in LUAD remain largely undefined. Using TCGA datasets and functional experiments in LUAD cell lines, we found that USP42 is significantly overexpressed in LUAD, and its high expression is associates with advanced stages and reduced overall survival. Besides, USP42 knockdown suppressed proliferation, colony formation, migration, invasion, and epithelial-mesenchymal transition (EMT). Mechanistically, USP42 physically interacted with PTGS2, thereby stabilizing PTGS2 from proteasomal degradation. Mouse Lewis lung carcinoma (LLC) cells with USP42 knockdown were subcutaneously injected into C57BL/6 mice. Our results demonstrated that USP42 depletion attenuated tumor growth, reduced PTGS2 and PD-L1 expression, alleviated CD8[+] T cell exhaustion, and increased intratumoral CD8[+] T cell infiltration. Importantly, forced PTGS2 expression largely rescued the tumor-suppressive phenotypes induced by USP42 knockdown, confirming PTGS2 as a critical downstream effector of USP2. In conclusion, our findings identify USP42 as a novel oncogenic deubiquitinase in LUAD that promotes tumor progression and immune escape by deubiquitinating and stabilizing PTGS2, and targeting USP42 is a promising strategy against LUAD.

To investigate the effects of platelet-rich fibrin on the biological properties of rabbit adipose-derived stem cells in vitro. Rabbit adipose-derived stem cells were isolated via enzymatic digestion, characterized by flow cytometry, and their trilineage differentiation potential was confirmed. Platelet-rich fibrin was prepared from rabbit blood and characterized by scanning electron microscopy. Adipose-derived stem cells were cultured with platelet-rich fibrin, and cell proliferation and migration were assessed using CCK-8, fluorescence staining, scratch would healing, and transwell assays. Isolated adipose-derived stem cells exhibited trilineage differentiation potential. Scanning electron microscopy revealed that the platelet-rich fibrin formed a three-dimensional fibrin network with platelets entrapped within the matrix. Functionally, platelet-rich fibrin significantly enhanced adipose-derived stem cell proliferation and migration. In the proliferation assay (CCK-8), platelet-rich fibrin treatment significantly increased cell numbers from day 3 onward (P < 0.05), reaching highly significant levels at day 5 and day 7 (P < 0.01) compared to controls. Notably, the direct co-culture group showed the highest proliferative activity at day 7, significantly exceeding the indirect co-culture group (P < 0.01). In the scratch wound healing assay, platelet-rich fibrin also significantly promoted adipose-derived stem cell migration (P < 0.05), with the direct co-culture group achieving near-complete wound closure by 24 h, particularly when cells were directly embedded within the three-dimensional platelet-rich fibrin scaffold. In this in vitro setting, platelet-rich fibrin demonstrates favorable biocompatibility with rabbit adipose-derived stem cells, significantly promoting both their proliferation (time-dependently) and migration, with the three-dimensional platelet-rich fibrin scaffold providing superior support compared to soluble factors alone. These findings indicate that the platelet-rich fibrin /adipose-derived stem cell composite possesses suitable cytocompatibility properties, warranting further in vivo investigation in rabbit urethral defect models to assess its regenerative potential.

Sepsis-induced myocardial dysfunction (SIMD) and related myocardial fibrosis (MF) contribute to poor cardiovascular outcomes. Whether the role of Nppb in this process remains unknown. This study aimed to investigate the effect of Nppb on MF in SIMD. Myocardial-specific Nppb conditional knockout (CKO) mice were generated using CRISPR/Cas9, and 8-week-old male mice were divided into control, sepsis, CKO control, and CKO sepsis groups. Cardiac function, histopathology, myocardial injury markers, endothelial-to-mesenchymal transition (EndMT)-related gene expression, and Notch1 signaling were measured. Nppb CKO alleviated myocardial injury, improved left ventricular systolic function, attenuated myocardial structural damage, suppressed EndMT, and increased survival in septic mice. Myocardial expression of Notch1 and Notch Intracellular Domain (NICD) was reduced in the LPS group but restored by Nppb CKO. In conclusion, myocardial Nppb deficiency protects against SIMD by maintaining cardiac contractility and structural stability, with associated changes in the Notch1 signaling pathway. Inhibiting Nppb may represent a potential therapeutic strategy for sepsis-related myocardial injury.

Gastric ulcer is a common gastrointestinal disorder associated with oxidative stress, inflammation, and disruption of the gastric mucosal barrier. This study investigated the gastroprotective and therapeutic effects of Phoenix dactylifera (Zaghloul) methanolic leaves extract against ethanol-induced gastric ulcer in male Wistar rats. Phenolic quantification of the extract revealed the presence of phenolic (2450.127 mg GAE/g) and flavonoid (2230.00 mg CE/g) contents. LC-ESI-MS-MS analysis led to detection of 23 compounds. Ethanol administration resulted in elevation of MDA, inflammatory modulators (TNF-α, ICAM-1 and IL-10) and reduction of TAC, GSH and cycloxygenase enzymes (COXI and COXII). Expression of E2F1, RAD21, NIBPL, and Vimentin genes was increased via ulcer induction. Treatment with the extract reversed ethanol effects by elevating TAC, GSH, COXI and COXII and reducing MDA, TNF-α, ICAM-1 and IL-10;TNF-α/IL-10 ratio was increased in the ulcerative group and reduced by treatment. Moreover, the extract modulated the expression of E2F1, RAD21, NIBPL, and Vimentin suggesting a potential association with attenuation of epithelial injury and mucosal repair. Histopathological examination demonstrated improvement of the mucosal architecture and mucin production, and reduction of fibrosis. Collectively, these findings demonstrate the impact of ulcer induction and extract treatment on the expression of the studied genes and support the gastroprotective and therapeutic effects of P. dactylifera leaves extract through antioxidant, anti-inflammatory, and mucosal healing activities. Further studies at the protein level using more dosages of the extract are needed to clarify the mechanistic significance of these gene expression changes and the appropriate dose of the extract.

Cisplatin is among the most widely used chemotherapeutic drugs for the treatment of solid tumors; nonetheless, nephrotoxicity persists as a significant dose-limiting adverse effect. The current study focuses on the protective effect of propolis against cisplatin-induced kidney damage in experimental rats. Nephrotoxicity was induced by a single intraperitoneal injection of cisplatin at a dose of 5 mg/kg, ip, followed by propolis oral administration. The chemical profile of the propolis extract was characterized using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Biochemical parameters of renal function (urea and creatinine), oxidative stress markers (MDA, SOD, CAT and GSH), proinflammatory markers (IL-6, TNF-α and IL-1ꞵ) and RT-PCR were performed along with histopathological and immunohistochemical investigations. LC-MS analysis revealed that the propolis contained several bioactive phenolic acids and flavonoids, with pyrocatechol, iso-ferulic acid, gallic acid, apigenin, kaempferol, quercetin, which may contribute to the antioxidant and nephroprotective effects of propolis. The results demonstrated that cisplatin markedly increased serum levels of urea and creatinine and elevated lipid peroxidation and inflammation, accompanied by notable damage in renal structure. Conversely, propolis treatment significantly ameliorated these adverse effects by restoring antioxidant status, reducing oxidative stress and improving histological structure. Furthermore, administration of propolis significantly suppressed inflammation by reducing proinflammatory cytokines levels and the expression of caspase-3. In addition, immunohistochemical results revealed that propolis treatment markedly increased Nrf2/HO-1 expression in kidney tissue. In conclusion, these findings suggested that propolis exerts nephroprotective effects against cisplatin-induced nephrotoxicity via its anti-inflammatory and antioxidant activities.

Statins, anti-hyperlipidemic drugs, are associated with skeletal muscle myopathy. New strategies to preserve muscle strength in statin-induced myopathy (SIM) are needed. This study assessed the effect of quercetin on simvastatin-induced myopathy in adult male rats, focusing on the modulation of autophagy-apoptosis crosstalk. Forty adult albino male rats were randomly divided into four groups. The control group (C) received distilled water, the quercetin group (Q) received quercetin (50 mg/kg/day, orally), the simvastatin (S) group received simvastatin (80 mg/kg/day, orally), and the quercetin + simvastatin (Q + S) group received both quercetin (50 mg/kg/day) and simvastatin (80 mg/kg/day, orally), all for 30 days. Gastrocnemius muscle contractility, muscle weight/tibial length ratio, histopathological examination, serum creatine kinase (CK) levels, oxidative stress markers, lipid profile, autophagy-related gene expression (LC3 and LAMP2), and apoptosis markers (BAX and caspase-3) were evaluated. Simvastatin induced muscle atrophy and significantly elevated serum CK levels. These changes were associated with increased oxidative stress, altered expression of autophagy-related genes (increased LC3 and decreased LAMP2), and increased expression of the apoptotic markers BAX and Caspase-3. Quercetin ameliorated these alterations by improving muscle contractility, reducing CK levels and oxidative stress, attenuating apoptosis, and modulating autophagy-related gene expression. Quercetin alleviated simvastatin-induced myopathy through its antioxidant and antiapoptotic effects and was associated with modulation of autophagy-related gene expression. These findings suggest that quercetin may represent a promising adjuvant strategy for mitigating statin-associated myopathy. Further studies evaluating autophagic flux at the protein level are warranted to clarify the underlying mechanisms.

Parkinson's disease (PD) is associated with systemic inflammatory and metabolic alterations that may extend beyond the central nervous system. Although 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) reproduces several pathological features of Parkinsonian neurodegeneration, its effects on pancreatic tissue remain poorly characterized. This study investigated MPTP-associated pancreatic histopathological and immunohistochemical alterations and evaluated the potential attenuating effects of Hexarelin. Fifty male BALB/c mice were randomly assigned to five groups (n = 10/group): Sham, MPTP, Hexarelin, PreHexarelin, and PostHexarelin. Pancreatic tissues were examined using hematoxylin and eosin staining and immunohistochemistry for amylin, β-amyloid, caspase-3, glucagon, insulin, CD11b, CD68, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). MPTP administration was associated with marked pancreatic injury characterized by acinar degeneration, cytoplasmic vacuolization, vascular congestion, interstitial edema, inflammatory cell infiltration, and degeneration of the islets of Langerhans. Immunohistochemical analysis demonstrated increased inflammatory (CD11b, CD68, IL-1β, and TNF-α) and apoptosis-associated (caspase-3) marker immunoreactivity, together with increased β-amyloid immunoreactivity and decreased insulin and amylin immunoreactivities. Hexarelin treatment attenuated these histopathological and immunohistochemical alterations in both treatment groups, although the magnitude of improvement varied among individual markers. These findings suggest that MPTP exposure is associated with pancreatic inflammation, apoptosis-associated changes, and alterations in endocrine-marker immunoreactivity. Hexarelin treatment was associated with attenuation of these pathological changes and partial preservation of islet morphology and endocrine-marker immunoreactivity; however, the PostHexarelin findings should be interpreted cautiously because the absence of a sequence- and time-matched post-MPTP vehicle control prevents complete separation of Hexarelin-associated effects from temporal changes after MPTP administration.

Ionizing radiation is widely used in the treatment of head and neck malignancies. However, its subacute effects on adjacent endocrine tissues have not yet been fully elucidated. This study aimed to investigate the structural changes that develop in the thyroid and parathyroid glands following irradiation and to evaluate HSP-90 immunoexpression in these tissues. The researchers initially randomized eighteen male Wistar-Hannover rats into control and irradiation groups (n = 9 per group). Rats in the irradiation group underwent 6-MV photon irradiation of the head and neck target region using a nominal prescribed single-fraction dose of 18 Gy with a Varian Clinac iX linear accelerator. One of the irradiated animals died on day 8; consequently, the day-21 analyses included nine control and eight irradiated animals. On day-21, the researchers performed histopathological examination and immunohistochemical evaluation of HSP-90 immunoexpression in thyroid and parathyroid tissues. Irradiation significantly increased vascular congestion (p = 0.031), inflammatory cell infiltration (p = 0.040), and epithelial desquamation (p = 0.006) in the thyroid tissue compared to the control group. It also significantly reduced HSP-90 immunoexpression in the thyroid follicular cells (p = 0.011). In contrast, irradiation did not cause marked disruption in the tissue architecture of the parathyroid glands. HSP-90 immunoexpression also showed no significant difference between the control and irradiation groups (p = 0.914). A single dose of high-energy photon irradiation induced subacute microstructural damage in the thyroid gland and reduced HSP-90 immunoexpression. The parathyroid gland, however, largely preserved its overall morphology and HSP-90 immunoexpression at day-21. These results indicate that thyroid and parathyroid tissues exhibit organ-specific subacute responses to irradiation.

Mitochondrial dysfunction and endoplasmic reticulum stress (ERS) play critical roles in the development of osteoarthritis (OA). Sirtuin 3 (SIRT3), a mitochondrial deacetylase, has regulatory effects on OA, but whether it can influence OA progression by modulating the crosstalk between mitochondrial dysfunction and ERS remains unclear. Therefore, the aim of this study was to investigate the effects of SIRT3 on mitochondrial dysfunction and ERS in OA. Anterior cruciate ligament transection was performed to establish an OA rat model, and C28/I2 chondrocytes were treated with IL-1β to establish an in vitro OA cell model for experimental investigation. Hematoxylin and eosin (H&E) staining, Safranin O/Fast Green staining, MitoTracker Red staining, and JC-1 staining were used to assess OA progression in rats and C28/I2 cell damage. RT-qPCR and Western blotting were used to measure the expression levels of target genes and proteins. Coimmunoprecipitation was utilized to examine protein interactions and acetylation levels. The result shows that SIRT3 expression was downregulated in OA rats and IL-1β-induced C28/I2 cells. SIRT3 overexpression significantly alleviated pathological damage to rat articular cartilage tissues, reduced chondrocyte loss, improved matrix metabolism in articular cartilage tissues, and ultimately mitigated OA progression in rats. Moreover, SIRT3 overexpression inhibited IL-1β-induced ERS in C28/I2 cells, reduced the activation of the unfolded protein response, and improved mitochondrial function. Additionally, the mitochondrial Ca2+ transport inhibitor Ru360 suppressed mitochondrial Ca2+ overload caused by disrupted endoplasmic reticulum calcium homeostasis and improved both mitochondrial function and ERS. DRP1 was upregulated in OA rats and IL-1β-stimulated cells, and changes in DRP1 acetylation signals together with decreased DRP1 protein abundance were observed following SIRT3 manipulation. Overall, SIRT3 manipulation was associated with acetylation signals of DRP1 and reduced DRP1 protein abundance, indicating that DRP1 is a potential mediator involved in the SIRT3‑linked restoration of mitochondrial function, attenuation of ERS, and alleviation of OA progression.

Mycoplasma ovipneumoniae (Mo) is a highly contagious pathogen that causes pneumonia in sheep, seriously threatening the sheep industry. Heat Shock Factor 1 (HSF1) is known to be important for host defense against lung infections, but its role in Mo infection was not understood. This study aimed to clarify HSF1 function during Mo infection. We created an in vivo infection model using Argali Hybrid Sheep and an in vitro model using alveolar macrophages. Immunohistochemistry was applied to check HSF1 expression and location in lung tissues. ELISA tests measured IL-6 and CXCL1 levels in serum and bronchoalveolar lavage fluid (BALF), while Western blot and RT-qPCR assessed HSF1, HSP70, and TLR2 expression in infected lungs and macrophages. Additionally, alveolar macrophages with overexpressed HSF1 were infected with Mo to study HSF1 regulation. Findings showed higher IL-6 and CXCL1 in BALF from infected sheep. Infected lung tissues and macrophages had lower HSF1 and HSP70 expression but higher TLR2 expression. Immunohistochemistry revealed less HSF1 in cell nuclei. Overexpressing HSF1 partly brought back HSP70 levels, reduced TLR2 increase, and improved Mo clearance. HSF1 overexpression enhances the immune response to Mo by controlling the HSP70/TLR2 pathway, offering a scientific foundation for strategies to prevent and control Mo infection.

Methotrexate (MTX) is a chemotherapeutic drug extensively used for the treatment of malignancies and autoimmune disorders. However, its clinical use is limited by cytotoxic effects on normal tissues, mediated by oxidative stress-induced depletion of antioxidant defenses and activation of apoptotic pathways. Rosmarinus officinalis (rosemary) contains bioactive compounds with established antioxidant and anti-apoptotic properties, indicating potential therapeutic effects against MTX-induced tissue injury. This study aimed to investigate the potential therapeutic effect of Rosmarinus officinalis in attenuating the cytotoxic effects of MTX on the submandibular salivary gland (SMSG) of male albino rats. This experimental animal study included twenty-eight adult male albino rats (aged 4-6 months), randomly allocated into four equal groups (n = 7): the control group received saline; the rosemary group received Rosmarinus officinalis methanolic extract (RME) (200 mg/kg, i.p.) once daily for 10 days; the MTX group received a single dose of MTX (40 mg/kg, i.p.) on day 1 only; and the MTX + rosemary group received MTX (40 mg/kg i.p.) on day 1, followed by RME (200 mg/kg i.p.) for 10 days. On day 11, the submandibular salivary glands were dissected and rinsed in 0.9% saline, and the right glands were divided into two parts: one preserved fresh for biochemical assessment of reduced glutathione (GSH), and the other fixed for histological and immunohistochemical analysis of cleaved caspase-3 and nuclear factor erythroid 2-related factor 2 (Nrf2). The control and rosemary groups exhibited normal glandular architecture, with the rosemary group showing the highest GSH concentration and Nrf2 expression. In contrast, the MTX group showed pronounced degenerative and atrophic changes in the acinar and ductal structures, coinciding with strong caspase-3 immunoreactivity, significantly reduced Nrf2 expression, and depleted GSH concentration (P ≤ 0.05). In the MTX + rosemary group, the glandular architecture showed partial improvement compared with the MTX group, associated with reduced caspase-3 immunoreactivity, recovered Nrf2 expression, and increased GSH concentration relative to the MTX group (P ≤ 0.05). Rosmarinus officinalis treatment significantly attenuated MTX-induced SMSG injury, as evidenced by enhanced Nrf2 immunoexpression, increased GSH concentration, and reduced cleaved caspase-3 expression. These findings suggest that rosemary exerts antioxidant and anti-apoptotic effects, contributing to the preservation of glandular integrity against MTX toxicity. Taken together, Rosmarinus officinalis may represent a promising adjunctive therapeutic strategy for attenuating MTX-induced glandular damage, warranting further investigation.

Bone transport-induced remote microinjury may promote diabetic foot ulcer healing but requires osteotomy and may cause fracture or osteomyelitis. This study evaluated whether osteotomy-free remote periosteal distraction (RPD) could promote repair in a rabbit ischemic diabetic foot wound model. Seventy-two diabetic male rabbits were randomized into RPD, Sham, and Control groups, and ischemic foot wounds were created. In the RPD group, a custom-made plate was implanted beneath the tibial periosteum and connected to an external distractor. The periosteum was distracted outward for 7 days and returned inward for another 7 days. Sham animals underwent the same implantation procedure without device adjustment, whereas Control animals received wound creation alone. Wound closure, histological changes, vascular parameters, and angiogenesis-related markers were evaluated. On days 8 and 16, RPD resulted in a smaller residual wound area than the Control and Sham groups. RPD increased epithelial thickness and collagen deposition compared with the Control and Sham groups. RPD was associated with increased serum VEGF levels, higher VEGF mRNA expression, and larger CD31-, VEGF-, and VEGFR2-positive areas in wound tissues. On day 16, ex vivo micro-CT angiography demonstrated higher vascular volume in the wound region after RPD treatment. Osteotomy-free RPD promoted wound repair in an ischemic diabetic wound model. These effects were associated with enhanced neovascularization and increased VEGF and VEGFR2 expression at the wound site. RPD may represent a potentially less invasive alternative to osteotomy-based tibial distraction for improving ischemic diabetic ulcer healing, but further mechanistic, safety, and clinical studies are required.