搜索结果：Journal of molecular histology

Cycloartane triterpenoids (CATT), natural compounds derived from Dysoxylum malabaricum, have substantiated promising anticancer potential, specifically against breast adenocarcinoma T-47D cells. However, the underlying mechanisms of action remain largely unexplored. The present work investigated the antiproliferative and pro-apoptotic effects of CATT on human breast cancer cells both in vitro and in vivo. CATT exhibited potent cytotoxicity against T-47D cells (IC50 = 40 µg/ml) while sparing normal cells (IC50 > 80 µg/ml). CATT treatment significantly suppresses the growth of breast cancer cells through G2/M phase cell cycle arrest, which was accompanied by decreased expression of cyclin B1. Interestingly, CATT increases intracellular reactive oxygen species (ROS) levels by disrupting mitochondrial dynamics, contributing to apoptosis induction. Further mechanistic studies revealed that increased levels of cleaved PARP-1, Bax, Cytochrome c, cleaved caspase-9 and cleaved caspase-3, along with decreased levels of PCNA, mutated p53 and Bcl-2, confirm mitochondrial-mediated cell death. Intriguingly, in vivo studies on Balb/c mice bearing T-cell lymphoma further supported these findings, showing extended lifespan, reduced tumour burden and restoring normal histoarchitecture. Importantly, molecular docking results corroborated the in vitro data by demonstrating direct interactions with apoptotic proteins and support the proposed apoptotic mechanism. In conclusion, CATT induces apoptosis in breast adenocarcinoma T-47D cells through mitochondrial dysfunction and caspase activation, and demonstrates potent antitumor activity both in vitro and in vivo. These findings highlight the potential of CATT as a promising natural compound for the development of novel breast cancer therapeutics. Schematic diagram of underlying anticancer mechanism induced by CATT in breast adenocarcinoma.

Granulomatous lobular mastitis (GLM) is a chronic, refractory inflammatory condition of the breast with an unclear etiology. Emerging evidence suggests a potential role of pyroptosis in the pathogenesis of GLM. This study investigated the therapeutic effects of Tuolitounong Decoction (TLTND) on pyroptosis in both in vitro and in vivo models of GLM. An in vitro GLM model was established using MCF10A mammary epithelial cells exposed to tissue homogenates derived from GLM-affected human breast tissue. Cells were treated with TLTND at varying concentrations, with or without the pyroptosis inhibitor necrosulfonamide. Pyroptosis-associated proteins, including caspase-1, gasdermin D (GSDMD), lipopolysaccharide-binding protein (LBP), and NLR family pyrin domain containing 3 (NLRP3), were assessed using immunofluorescence and western blotting. Cell viability was evaluated using the Cell Counting Kit-8 (CCK-8) assay. For in vivo analysis, a rat GLM model was induced by injecting a combination of human GLM tissue homogenate supernatant and Freund's complete adjuvant into the third and fourth pairs of mammary glands in female Sprague Dawley rats. TLTND was administered via daily oral gavage for 21 days. Post-treatment evaluations included histopathological assessment, expression of estrogen receptor (ER) and progesterone receptor (PR), and pyroptosis-related biomarkers in mammary gland tissue. Exposure to GLM homogenate successfully induced pyroptosis-related pathological features in MCF10A cells, whereas serotonin (5-HT) inhibition showed no significant effect. TLTND treatment demonstrated dose- and time-dependent effects, with enhanced therapeutic efficacy at 10 mg/mL after 48 h (P = 0.009). Of note, TLTND significantly reduced the protein expression levels of key pyroptosis markers (cleaved caspase-1, n-GSDMD, cleaved IL-18 and cleaved IL-1β) by 52.6%, 91.0%, 78.1% and 87.1%, respectively, compared to the model group in rat mammary tissue. Co-treatment with TLTND and necrosulfonamide significantly downregulated the expression of caspase-1, GSDMD, LBP, and NLRP3, while increasing cell viability. In vivo, both prednisone acetate and TLTND ameliorated histopathological features of GLM. Notably, TLTND promoted localized abscess maturation and increased ER and PR expression, while reducing levels of interleukin-1β and GSDMD-N in mammary tissue. TLTND demonstrated therapeutic potential in reducing pyroptosis in mammary epithelial cells and modulating inflammation in granulomatous lobular mastitis, partly by downregulating the caspase-1/GSDMD-mediated pyroptosis signaling pathway. These findings support further investigation of TLTND as a complementary treatment approach for GLM and provide insight into its underlying mechanisms of action.

Focal cortical dysplasia (FCD) is a leading cause of drug-resistant epilepsy, whereas its molecular and cellular mechanisms remain poorly understood. This study aimed to characterize the cellular heterogeneity of FCD and investigate the function of ferroptosis in FCD pathogenesis. Single-nucleus RNA sequencing was carried out on epileptogenic cortical tissues from 18 patients with FCD and 6 perilesional control samples with normal histology. Data were analysed using uniform manifold approximation and projection for dimensionality reduction and visualization. Differentially expressed genes (DEGs) were identified and subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. UCell scoring and gene set enrichment analysis (GSEA) were applied to assess pathway activity. Expression levels of ferroptosis-related genes (FRGs) were validated by immunofluorescence, and biochemical assays quantified the levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) and lipid peroxides (LPO). A total of 170 747 nuclei were profiled, resolving five major cell types, including inhibitory neurons, excitatory neurons, astrocytes, microglia and oligodendrocytes. DEGs across these populations were significantly enriched in ferroptosis and oxidative stress-associated pathways. UCell and GSEA highlighted remarkable alterations in ferroptosis, apoptosis and oxidative stress, particularly in inhibitory neurons and astrocytes. Immunofluorescence confirmed upregulation of key FRGs, including ferritin light chain, ferritin heavy chain 1, poly rC binding protein 1, microtubule-associated protein 1 light chain 3B and prion protein-encoding gene, in FCD tissues. Concordantly, biochemical assays demonstrated reduced SOD and GSH levels, alongside elevated MDA and LPO levels, confirming the transcriptional and histological findings. The results indicated that ferroptosis may play a notable role or act as a concurrent mechanism in the pathogenesis of FCD, potentially contributing to the neuronal and glial dysfunction and epileptogenesis. Integrating transcriptomic, histological and biochemical data, this study demonstrated that targeting ferroptosis-related pathways may hold promise as a potential therapeutic strategy for FCD, providing new insights into the molecular mechanisms underlying this condition. This study pioneers the first single-nucleus transcriptomic atlas for Focal Cortical Dysplasia (FCD) types I and II, deciphering the cellular heterogeneity across five major brain cell types within the epileptogenic cortex. Through integrated multi-omics analysis, it reveals for the first time a significant association between the ferroptosis pathway and FCD pathogenesis. We identify and validate ferroptosis-related genes (e.g., FTH1, FTL, PCBP1) as potential biomarkers and therapeutic targets, supported by congruent biochemical evidence of oxidative stress in this drug-resistant epilepsy.

The introduction of molecular diagnostics has completely changed oncology, moving the classification based on histology to biomarker-driven precision medicine. Through the use of biomarkers genomic, transcriptomic, proteomic, metabolomic, and epigenetic changes the cancer field is able to gain much needed insights for detection at an early stage, prognosis that is accurate, and treatment selection that is personalized. The current paper looks at the high-tech molecular processes that make the discovery and use of biomarkers possible, including next-generation sequencing, mass spectrometry, and PCR-based techniques, which are capable of very sensitive detection of everything from single-nucleotide variants to complete proteomic profiles. The talk points out the successful clinical translations like EGFR inhibitors in lung cancer and PARP inhibitors in BRCA-mutant cancers, at the same time acknowledging the biggest hurdles in biomarker validation such as standardization, ethical issues, and making sure no one is left out in access to the benefits. Some of the most promising new technologies like single-cell sequencing, CRISPR-based diagnostics, and artificial intelligence are expected to bring about further revolution in the area by unraveling the complexities of tumor heterogeneity, facilitating super-sensitive detection, and assimilating data from multiple omics. The future of cancer biomarker analysis is in the application of diverse molecular layers to evolve dynamic and comprehensive profiles of individual tumors. This will enable real-time monitoring through liquid biopsy and point-of-care devices. Ultimately, the constant improvement and the interdisciplinary partnership in biomarker science are of utmost importance to the full realization of the personalized oncology's promise and the enhancement of the survival rate of all cancer patients.

The aim of this study is to evaluate the clinical efficacy of Fufang Zaofan Pills in individuals diagnosed with lower-risk myelodysplastic syndrome (MDS) and to assess associated changes in peripheral blood dendritic cell (DC) levels. A cohort of 60 individuals with newly diagnosed lower-risk MDS were enrolled at the Department of Hematology, Jiading Central Hospital, affiliated with Shanghai University of Medicine & Health Sciences between February 2022 and July 2023. Participants were randomly allocated into two groups: an integrated Chinese-Western medicine group (n&#x2009;=&#x2009;40) and a Western medicine group (n&#x2009;=&#x2009;20). Both groups received standard Western medical therapy as per clinical guidelines, while the integrated group received additional oral administration of Fufang Zaofan Pills. Following the treatment course, clinical outcomes were compared between groups, including Western medicine efficacy, changes in traditional Chinese medicine (TCM) syndrome scores, and peripheral blood DC levels before and after treatment. At baseline, lower-risk MDS patients exhibited significantly elevated levels of DC and mDC compared to healthy controls (p&#x2009;<&#x2009;0.05). The objective response rate (ORR) in the integrated Chinese-Western medicine group was 47.5%, which was significantly higher than the 20% observed in the Western medicine group (p&#x2009;<&#x2009;0.05). The total effective rate for TCM syndrome scores was also significantly greater in the integrated group (80% vs. 60%, p&#x2009;<&#x2009;0.05). Post-treatment analysis indicated significant changes in DC percentage, absolute count, and mDC-related indices in the integrated group compared to baseline values (p&#x2009;<&#x2009;0.05), with intergroup comparisons demonstrating statistically significant differences in favor of the integrated therapy (p&#x2009;<&#x2009;0.05). The addition of Fufang Zaofan Pills to standard treatment appears to enhance clinical efficacy in lower-risk MDS and is associated with modulation of peripheral blood DC levels. These preliminary findings contribute evidence supporting further investigation into the integration of TCM and Western medical approaches in the management of lower-risk MDS, particularly regarding their immunomodulatory potential.

Sleep amount and quality are important to maintain health, light exposure transported to the pineal gland via suprachiasmatic nucleus pineal pathway, and this affect melatonin synthesis. Melatonin exerts its action by receptors called MT1, MT2 that found in peripheral tissues including pancreas, which affect its endocrine function. In this study we investigate the effect of type of sleep disturbance on the histological structure and the expression of melatonin receptor (MTNR1B) in the pancreas. A sample of 45 adult healthy male rats had free access to water and feeding, divided into 3 groups (15 in each group): the control group had normal 24 h diurnal variation, group A subjected to interruption of sleep by light exposure for 2 h at three intervals, and group B subjected to a reduction in sleep duration by 7 h. This experiment continued for 14 days, animals were scarified by euthanasia by cervical dislocation, pancreatic tissue prepared for paraffin blocks and stained by H&E, and IHC for MTNR1B. Histomorhometric analysis done by image J (V 1.54), immunohistochemical assessment done by Aprioscope image analysis software 12.4.6.5003) mean total positivity of expression was selected, statistical analysis done by SSPS (25 V) and Tukey&#xa78c;s test. Histological evaluation revealed changes in sleep disturbance groups including fat deposition, vascular dilatation, and apoptotic changes for group of cells within islets of Langerhans which showed significant reduction in their count in sleep disturbance groups compared to control, and non-significant change in their area across the groups, but significant reduction in area in sleep deprived group compared to sleep interruption. Immunohistochemical expression of MTNR1B showed it only expressed within the area of islets of Langerhans, with non-significant increase in expression in sleep disturbed groups, being more in sleep deprived group. This study showed that the pancreas is affected by sleep disturbance patterns, mainly its endocrine part (islets of Langerhans), with study expression of melatonin receptor within them. This may suggest a role of melatonin in maintaining pancreatic &#xa7b5; cells.

Retinal detachment (RD) remains an ophthalmologic emergency with high anatomical success rates after surgery but often suboptimal visual outcomes. This study aimed to identify transcriptomic signatures linked with clinical severity in human RD to uncover the molecular basis of variability in functional recovery. Full-length RNA sequencing (RNA-seq) was performed on freshly collected human retinas from patients with rhegmatogenous RD. Principal component analysis was used to derive a composite severity framework, which guided subsequent analysis (differential gene expression, protein-protein interaction, multivariable modeling, and functional enrichment) to identify potential biomarkers and pathways associated with disease severity. Transcriptomic changes were primarily driven by a core severity axis, highlighting baseline best-corrected visual acuity and macular/foveal involvement as clinically interpretable proxies of severity. Severe RD was characterized by strong upregulation of immune and inflammatory genes and pathways, along with activation of Rho-GTPase pathways and G protein-coupled receptors-signaling, suggesting an active immune microenvironment. Consistent downregulation of metabolic and photoreceptor associated pathways, reflecting mitochondrial dysfunction and bioenergetic failure, was also observed. Transcriptomic shifts seemed to occur beyond clinically relevant severity thresholds rather than along linear gradients. PTPRC, FCGR3A, and SCARB1 emerged as central hub proteins with potential biomarker value. Unexpected enrichment of sensory and olfactory receptor pathways suggested a potential contribution to post-detachment neurodegeneration. Individual variables largely recapitulated these transcriptional signatures, reinforcing their applicability in stratification. Inflammation, immune dysregulation, and metabolic impairment emerged as key molecular indicators of severe RD, supporting the development of molecular-based stratification and potential adjuvant therapies.

Prostate cancer (PCa) is among the most frequently diagnosed malignancies in aging men and remains a major public health concern, highlighting the need for effective and safe preventive strategies. Annona crassiflora Mart. seed extract (ASE) was evaluated using the transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Animals were divided into control and ASE-treated groups evaluated from 8-12 to 12-16&#xa0;weeks of age. Control groups received the vehicle (water containing 10% DMSO), whereas ASE-treated groups received 100&#xa0;mg/kg body weight of ASE. The preservation of hepatic histoarchitecture and normal body weight gain demonstrated that ASE was well tolerated and did not show hepatotoxic effects. ASE treatment increased the frequency of healthy prostatic epithelium in early prostate lesions (predominantly low-grade prostatic intraepithelial neoplasia (LGPIN)) and late prostate lesions (high-grade prostatic intraepithelial neoplasia (HGPIN) and well-differentiated adenocarcinoma (WDAC)) of PCa. Furthermore, ASE reduced HGPIN incidence in the ASE 12-16 group, showing a lower incidence of WDAC. In addition, ASE reduced androgen receptor (AR) protein levels as well as cytoplasmic and nuclear AR localization in late prostate lesions (HGPIN and WDAC) of PCa. ASE also induced apoptosis through multiple pathways, including upregulation of BID, BAD, and BAX, activation of caspase-8, caspase-9, and caspase-3, and downregulation of BCL-2, BCL-xL, and MCL-1. Thus, ASE promotes a pro-apoptotic environment unfavorable to tumor survival. Taken together, these results reinforce the potential of ASE to modulate prostatic homeostasis, indicating a promising strategy with chemopreventive effects and potential as an adjuvant therapy against PCa.

Prostate cancer (PCa) is among the most frequently occurring cancers in men. Its occurrence and progression are closely related to metabolic reprogramming, with abnormally active glycolysis being a notable feature sustaining rapid proliferation. Recent years have seen growing interest in the regulatory role of epigenetic modifications, particularly N&#x2076;-methyladenosine (m6A) RNA methylation. AlkB homolog 5 (ALKBH5), as an m6A demethylase, may influence glycolysis in PCa by regulating target genes, offering new directions for mechanistic studies and therapy. The impact of different concentrations of Icariin (ICA) on PCa cell viability was first assessed using Cell Counting Kit-8 assays. Colony formation and Transwell assays were used to evaluate the effects of ICA on cell proliferation and migration, respectively. Apoptosis was detected by Annexin V-fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) double staining. Glycolysis was evaluated by measuring the extracellular acidification rate (ECAR), oxygen consumption rate (OCR), and levels of glucose, lactate, and adenosine triphosphate (ATP). Methylated RNA immunoprecipitation-PCR was used to assess the effect of ICA on m6A methylation. The messenger RNA (mRNA) and protein expression of methyltransferase-like 3 (METTL3), METTL14, WTAP, fat mass and obesity-associated protein (FTO), and ALKBH5 were detected by quantitative real-time PCR and Western blot, respectively. Stable ALKBH5-knockdown and glutamyl-tRNA synthetase 2 (EARS2)-overexpressing PCa cell lines were constructed for functional studies. The half-life of EARS2 mRNA after ALKBH5 knockdown was determined using an actinomycin D assay to evaluate mRNA stability. RNA immunoprecipitation (RIP) experiments were performed to verify the interaction between ALKBH5 and EARS2 mRNA. In addition, a xenograft model was established by injecting PCa cells for in vivo validation. Immunohistochemistry (IHC) was used to detect protein expression in tumor tissues. ICA inhibited the proliferation (reduced cell viability, decreased colony formation), metastasis (impaired migration and invasion), and promoted apoptosis of PCa cells in a concentration-dependent manner, with no significant effect on cell viability at concentrations below 25 &#xb5;M. ICA also concentration-dependently suppressed glycolysis in PCa cells. Treatment with 25 &#xb5;M ICA reduced the global m6A methylation level and specifically upregulated the mRNA and protein expression of the demethylase ALKBH5, while the expression of other m6A-related regulatory factors was not significantly affected. Functional experiments showed that ALKBH5 knockdown or EARS2 overexpression promoted PCa cell proliferation, metastasis, and glycolysis (increased ECAR, glucose uptake, lactate production, ATP levels, and glycolysis-related protein expression; decreased OCR), and attenuated the inhibitory effects of ICA on these processes. RIP experiments confirmed that ALKBH5 interacts with EARS2 mRNA and that ALKBH5 reduces EARS2 mRNA stability via m6A demethylation, thereby downregulating its expression. In vivo, ICA effectively inhibited tumor growth in mice. IHC results showed that ICA upregulated ALKBH5 expression and downregulated the expression of hexokinase 2, lactate dehydrogenase A, and pyruvate kinase M2 in tumor tissues. These inhibitory effects of ICA were weakened by ALKBH5 knockdown. ICA suppresses the progression of PCa by inhibiting glycolysis, a process in which upregulation of ALKBH5 mediates m6A demethylation of EARS2 mRNA, leading to its destabilization and decreased expression. These findings were further validated in vivo.

Reduced response to targeted therapy in human epidermal growth factor receptor 2 (HER2)-positive breast cancer underscores the need for novel therapeutic strategies and molecular targets. Cluster of differentiation 24 (CD24), an oncogenic molecule closely associated with aggressive tumor phenotypes, was investigated for its expression in HER2-positive breast cancer and its potential as a candidate target for combination therapy. Herein, we explored the expression of CD24 in breast cancer (BC) and its prognostic significance. In HER2-positive BC cells, CD24 was knocked down to assess its impacts on tumor biological behaviors and synergistic effects with trastuzumab. Furthermore, the underlying mechanism of CD24 was elucidated using a series of experimental assays, including Western blotting (WB), immunohistochemistry (IHC), terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, and mRFP-GFP-LC3 dual-fluorescence autophagy assay. In vivo experiments were conducted to evaluate the therapeutic efficacy of CD24 inhibition combined with trastuzumab. CD24 was highly expressed in HER2&#x2009;+&#x2009;breast cancer and associated with poor prognosis. CD24 knockdown significantly suppressed the proliferation, migration, and invasion of HER2-positive BC cells. Moreover, CD24 depletion promoted apoptosis and autophagy in HER2-positive breast cancer, accompanied by the inactivation of the Akt/mammalian target of rapamycin (mTOR) signaling pathway. In vivo data demonstrated that the combination regimen of CD24 knockdown and trastuzumab markedly reduced tumor burden. CD24 silencing modulates autophagy and induces apoptosis via Akt/mTOR inhibition, and its combination with trastuzumab exerts potent antitumor effects, offering a promising strategy for HER2&#x2009;+&#x2009;breast cancer.

The WNK3-SPAK-NKCC1 signaling pathway has been implicated in the pathogenesis of brain injury. The aim of this study is to examine the involvement of this pathway in intracerebral hemorrhage (ICH) and evaluate the therapeutic potential of acupuncture in modulating its activity. A total of 210 Sprague Dawley rats were randomly assigned to experimental groups. ICH was induced in all groups except the sham group via autologous blood injection. Point-through-point acupuncture was administered on the Baihui (GV20) and Qubin (GB7) acupoints. Protein expression levels within the WNK3-SPAK-NKCC1 pathway, as well as inflammatory and apoptotic markers in the perihematomal region, were assessed using western blotting, immunohistochemistry, immunofluorescence co-localization, and enzyme-linked immunosorbent assay. Brain water content, hematoxylin-eosin staining, and neurological function scoring were used to evaluate histopathological changes and functional outcomes. Acupuncture significantly improved neurological function, alleviated cerebral edema, and reduced perihematomal pathological injury in ICH rats at all observed time points. Notably, the therapeutic effect was most pronounced after seven consecutive acupuncture treatments. Time-course analysis of WNK3 and cleaved caspase-3 expression, together with correlation analysis, revealed that the neuroprotective effects of acupuncture were potentially associated with WNK3-mediated apoptotic mechanisms. Based on these findings, day 7 was identified as the optimal time point for further mechanistic investigation. At this time point, continuous acupuncture treatment significantly improved neurological function and decreased brain water content. In addition, the expression levels of WNK3, phosphorylated SPAK and NKCC1, TNF-&#x3b1;, and cleaved caspase-3 were significantly reduced, accompanied by decreased NeuN/TUNEL co-localization (p&#x2009;<&#x2009;0.05), suggesting that acupuncture may exert neuroprotective effects partly through inhibition of the WNK3-SPAK-NKCC1 signaling pathway. Acupuncture may alleviate brain injury following ICH by suppressing activation of the WNK3-SPAK-NKCC1 signaling pathway, thereby decreasing cerebral edema, inflammatory responses, and neuronal apoptosis.

Gestational diabetes mellitus is a common metabolic disorder of pregnancy associated with maternal metabolic disturbances, placental dysfunction, and adverse fetal outcomes. However, data on placental histopathological and apoptotic alterations in experimental models remain limited. This study evaluated the protective effects of an ethanolic extract of propolis on metabolic, hematological, oxidative, and placental structural changes in a rat model of diabetes induced during gestation using streptozotocin. Pregnant rats were assigned to four groups: a control group, a control group treated with an ethanolic extract of propolis, a diabetic group, and a diabetic group treated with the ethanolic extract of propolis. Diabetes during pregnancy was induced by a single intraperitoneal injection of streptozotocin (35&#xa0;mg/kg). Metabolic parameters, hematological indices, oxidative stress markers, and placental histology were assessed. Diabetic rats showed pronounced hyperglycemia, reduced maternal weight gain, dyslipidemia, renal dysfunction, and hematological alterations consistent with anemia and inflammation. Placental examination revealed structural disorganization, trophoblastic degeneration, fibrosis with focal blood accumulation, and increased oxidative stress, evidenced by enhanced lipid peroxidation and decreased antioxidant defenses. Increased expression of caspase-3 indicated enhanced apoptotic activity. Treatment with the ethanolic extract of propolis significantly improved metabolic and hematological parameters, restored antioxidant status, and attenuated placental damage. It also reduced caspase-3 expression, suggesting decreased apoptosis. In conclusion, propolis exerts protective effects against placental injury associated with diabetes induced during gestation and may represent a promising natural therapeutic approach.

Ovarian cancer is among the most lethal gynecologic malignancies worldwide, with high mortality rates largely attributable to late diagnosis and the development of therapeutic resistance. Cellular proteostasis in cancer cells is tightly regulated by the ubiquitin-proteasome system (UPS) and autophagy, two major protein quality control pathways. Key proteins involved in these pathways, including p97/valosin-containing protein (p97/VCP), ubiquitin (Ub), p62/SQSTM1, and LC3B, have been implicated in cancer progression; however, their coordinated regulation in ovarian cancer remains incompletely understood. In this study, we investigated the expression and regulation of UPS- and autophagy-related proteins in the human ovarian cancer cell line MDAH-2774 under pharmacological modulation of autophagy. Cells were treated with rapamycin (10&#xa0;&#xb5;M), an autophagy inducer, or chloroquine (50&#xa0;&#xb5;M), an autophagy flux inhibitor, for defined incubation periods. Protein expression levels and cellular localization of p97/VCP, Ub, p62, and LC3-II were analyzed using Western blotting, immunofluorescence staining, and siRNA-mediated p97/VCP silencing. Our results demonstrated that p97/VCP and ubiquitin were expressed at relatively high levels in ovarian cancer cells, whereas autophagy markers p62 and LC3B showed reduced basal expression, suggesting dysregulated autophagic activity. Rapamycin treatment markedly increased p97/VCP expression, supporting its involvement in autophagy induction. In contrast, chloroquine treatment significantly reduced p97/VCP levels while inducing a pronounced accumulation of Ub, p62, and LC3-II (p&#x2009;<&#x2009;0.05), consistent with impaired autophagic flux and disrupted proteasomal degradation. Furthermore, siRNA-mediated suppression of p97/VCP significantly decreased the expression of p97/VCP, LC3-II, ubiquitin, and the autophagy-modulating responses to rapamycin and chloroquine compared with control siRNA (p&#x2009;<&#x2009;0.05). p97/VCP suppression was associated with alterations in endoplasmic reticulum-associated degradation (ERAD) and a concomitant reduction in unfolded protein response (UPR)-related protein levels, indicating increased cellular stress. Collectively, these findings highlight p97/VCP as a central regulator of proteostasis in ovarian cancer through its coordinated roles in both the UPS and autophagy pathways. Dysregulation or pharmacological inhibition of p97/VCP may exacerbate proteotoxic stress by disrupting the balance between autophagy and UPR signaling, potentially contributing to tumor progression and therapy resistance. Further mechanistic studies are warranted to elucidate the underlying signaling networks and to evaluate p97/VCP-mediated proteostasis pathways as potential therapeutic targets in ovarian cancer.

Hypertrophic scars are prevalent fibrotic disorders that arise following skin wound healing, often resulting in both functional impairments and significant cosmetic concerns for affected individuals. Current therapeutic options, with pressure therapy being standard, frequently encounter challenges related to patient compliance and inconsistent efficacy. Given these limitations, there is a compelling need for innovative approaches to improve scar management in clinical settings. This study aimed to systematically evaluate the safety and therapeutic efficacy of a novel pressure-enhanced scar patch designed to address the aforementioned clinical challenges. A rabbit ear hypertrophic scar model was utilized, established through the perichondrium-preservation method, which demonstrated a 100% modeling success rate. The experimental subjects were stratified into pressure-treated and control groups, with multidimensional assessments conducted at 10, 20 and 30&#xa0;days post-operation to gauge the outcomes. Safety evaluations indicated a cumulative irritation index of 0.33 from skin irritation tests (score&#x2009;<&#x2009;0.5), confirming non-irritancy. Infrared thermal imaging revealed no significant alterations in skin temperature compared to baseline (P&#x2009;>&#x2009;0.05), affirming that local blood supply remained unaffected. Histological examinations, including Hematoxylin and Eosin (HE) and PTAH staining, corroborated the absence of epidermal damage, tissue edema, and inflammatory cell infiltration. Efficacy assessments showed a significant reduction in the scar elevation index (SEI) from 3.5&#x2009;&#xb1;&#x2009;0.31 at day 10 to 1.9&#x2009;&#xb1;&#x2009;0.44 at day 30 in the treatment group, alongside marked declines in microvessel density (MVD) and collagen volume fraction (CVF), with all parameters achieving statistical significance (P&#x2009;<&#x2009;0.05). The findings of this study substantiate the high safety profile and significant therapeutic efficacy of the pressure-enhanced scar patch. It effectively mitigates scar pathology by inhibiting fibroblast activation, reducing angiogenesis, and collagen deposition while promoting the reconstruction of elastic fibers. These results provide a reliable foundation for the clinical management of hypertrophic scars and pave the way for further research into optimized scar treatment strategies.

Vitiligo is a pigmentary disorder characterized by the loss of functional melanocytes, leading to depigmented skin patches and significant psychological burden. Platelet-rich plasma (PRP) and its derived exosomes have recently gained attention as potential regenerative therapies due to their immunomodulatory and antioxidant effects.To evaluate the therapeutic effects of platelet-rich plasma (PRP) versus its derived exosomes in a hydrogen peroxide-induced depigmentation model in guinea pigs.Forty-two adult male guinea pigs were divided into control, depigmentation, PRP-treated, and PRP-exosome-treated groups. Depigmentation was induced using topical H2O2. A recovery group was included to assess spontaneous repigmentation after discontinuation of H2O2. Histological, immunohistochemical, histomorphometric, and biochemical analyses were performed.The depigmentation group showed progressive spongiosis in the epidermis with significant increase in the epidermal thickness. Significant decrease in melanin pigment and significant decrease in the number of melanocytes were noticed in the epidermis and hair follicles. Administration of PRP and PRP-Exosomes reversed these histological alterations with significant reduction in epidermal thickness and negative CD3 reaction in the dermis. More improvement was observed in PRP-exosome group compared to PRP group, as evidenced by significant increased amount of melanin granules and number of melanocytes in exosome group.Within the limitations of this oxidative stress-induced depigmentation model, both PRP and its derived exosomes were associated with histological improvement, with greater changes observed in the exosome-treated group. However, further studies are needed to confirm these findings in models more representative of human vitiligo.

Neuropathic pain (NPP) is a chronic and refractory condition resulting from injury or disease of the nervous system. This study aims to elucidate the molecular mechanism by which TGF-&#x3b2;1 alleviates neuropathic pain through modulation of the CXCL1/CXCR2 signaling pathway. A rat model of neuropathic pain was established via spinal nerve ligation (SNL). Mechanical allodynia and thermal hyperalgesia were assessed using electronic von Frey filaments and the plantar test, respectively. Histopathological changes, cellular apoptosis, and levels of pro-inflammatory cytokines (IL-1&#x3b2;, IL-6, TNF-&#x3b1;) in spinal cord tissues were evaluated using hematoxylin and eosin (H&E) staining, TUNEL assay, and enzyme-linked immunosorbent assay (ELISA). Bioinformatics analysis (based on the GSE24982 dataset), quantitative real-time PCR (qRT-PCR), Western blotting, immunofluorescence staining, and dual-luciferase reporter assays were employed to investigate the regulatory role of TGF-&#x3b2;1 in the CXCL1/CXCR2 pathway. The direct inhibitory effect of TGF-&#x3b2;1 on CXCL1 expression was confirmed using primary astrocyte cultures, while intrathecal administration of a CXCL1 neutralizing antibody or the CXCR2 antagonist SB225002 was used to assess the functional impact of pathway blockade. SNL induced significant mechanical allodynia and thermal hyperalgesia, accompanied by spinal cord tissue damage, increased neuronal apoptosis, and elevated levels of pro-inflammatory cytokines. Expression of CXCL1 and CXCR2 was markedly upregulated following SNL. Double-immunofluorescence staining and quantitative co-localization analysis showed that CXCL1 signals were associated with GFAP-positive regions, whereas CXCR2 signals were associated with NeuN-positive regions, suggesting possible involvement of the CXCL1/CXCR2 axis in glia-neuron signaling. In primary astrocytes, TGF-&#x3b2;1 treatment was associated with SMAD2/3 signaling and reduced CXCL1 promoter activity and expression. In vivo, TGF-&#x3b2;1 attenuated pain-related behaviors, apoptosis, and neuroinflammation, whereas CXCL1 administration partially reversed these protective effects. TGF-&#x3b2;1 attenuates SNL-induced neuropathic pain at least partly by suppressing CXCL1 expression and modulating a CXCL1/CXCR2 axis that may be involved in glia-neuron signaling.

Both platelet-rich plasma (PRP) and exosomes derived from bone marrow mesenchymal stem cells (BMSCs) exhibit therapeutic potential for spinal cord injury (SCI). However, the combined application of PRP and BMSCs-derived exosome (PRP-primed BMSCs-EXO, PRP-BMSCs-EXO) has not been reported. This study aimed to investigate whether the combined use confers superior therapeutic effects and to explore the underlying molecular mechanisms. First, we examined the effects of PRP priming on the proliferation and exosome secretion of BMSCs. High-throughput sequencing and RT-qPCR were used to analyze the differential miRNA expression profiles between exosomes from PRP- primed BMSCs (PRP-BMSCs-EXO) and naive BMSCs (BMSCs-EXO). Subsequently, a series of in vitro SCI models were established to evaluate the therapeutic effects of PRP-BMSCs-EXO on SCI. TargetScan, RNAhybrid, GO, KEGG, dual-luciferase reporter assay, RT-qPCR, western blotting (WB), and rescue experiments were performed to explore the roles of miRNAs in PRP-BMSCs-EXO and the associated signaling regulatory mechanisms in SCI. Finally, a series of in vivo SCI models were used to verify the therapeutic efficacy and mechanisms of PRP-BMSCs-EXO in vivo. PRP priming promoted the proliferation and exosome secretion of BMSCs. High-throughput sequencing and RT-qPCR revealed that multiple miRNAs were highly expressed in PRP-BMSCs-EXO compared with BMSCs-EXO, among which miR-29a-3p was the most abundant. In in vitro SCI models, PRP-BMSCs-EXO exerted better therapeutic effects than BMSCs-EXO and PRP. Mechanistically, PRP-BMSCs-EXO may enhance the viability and migration and inhibit the apoptosis and autophagy of pheochromocytoma 12 (PC12) cells by delivering miR-29a-3p, thereby promoting nerve regeneration and SCI repair. Bioinformatic analysis and luciferase assays indicated that PTEN may be a downstream target gene of miR-29a-3p. Rescue experiments confirmed that the miR-29a-3p/PTEN/PI3K/Akt/mTOR axis may mediate the regulatory effects of PRP-BMSCs-EXO on neuronal apoptosis, autophagy, and nerve regeneration in SCI. These findings were further validated in in vivo SCI models. Compared with PRP and BMSC-derived exosomes alone, PRP-primed BMSC-derived exosomes provide superior neuroprotective effects against SCI. PRP-BMSCs-EXO inhibits neuronal apoptosis and autophagy and promotes nerve regeneration via the miR-29a-3p/PTEN/PI3K/Akt/mTOR axis, representing a promising and effective therapeutic strategy for SCI.

Periodic fasting is known to improve metabolic health, but its impact on pancreatic islet plasticity remains unclear. We investigated the effects of intermittent fasting-mimicking diet (FMD) cycles on islet architecture and function in mice by performing immunohistochemical, ultrastructural, and metabolic analyses after fasting and after refeeding separately. Twelve-week-old female C57BL/6J mice were randomized to fasting (n&#x2009;=&#x2009;9), refeeding (n&#x2009;=&#x2009;10), or control group (n&#x2009;=&#x2009;10). FMD was supplied weekly for 3&#x2009;days (50%, 10%, 10% of daily caloric intake) followed by 4&#x2009;days of chow food ad&#xa0;libitum (except for pre-IGTT food withdrawal and the duration of the IGTT). Intraperitoneal glucose tolerance tests (IGTTs) were performed at day 11 (fasting group), day 14 (refeeding group), and day 13 (control group). Mice were sacrificed 7&#x2009;days after IGTT, and pancreata were subjected to fluorescence immunohistochemistry or scanning electron microscopy (STEM). Bodyweight, blood glucose, proinsulin, and IGF-1 concentrations were significantly decreased after fasting but rebounded after refeeding. Pancreatic insulin+glucagon+, BRN4+, and PDX1+BRN4+ cells increased significantly after fasting and tended to remain high after refeeding, thereby indicating increased pancreatic islet plasticity after fasting. In STEM images, the insulin granule core-to-halo ratio increased significantly after fasting. The fasting but not the refeeding group showed impaired glucose tolerance. The more crystallized mature &#x3b2;-cell granules indicate increased insulin secretory capacity, and the reduced proinsulin-to-insulin ratio suggests reduced endoplasmic reticulum stress in &#xdf;-cells after fasting. We propose that this observed plasticity may provide a basis for novel concepts of in&#xa0;vivo &#x3b2;-cell regeneration. However, further studies to investigate molecular mechanisms of fasting/refeeding in murine type 1 diabetes to evaluate its therapeutic potential are needed.

Vaginal self-sampling is increasingly integrated into cervical cancer screening programs, requiring evidence of accurate HPV testing on self-collected samples. This study evaluated the clinical accuracy of Allplex HPV HR Detection (Allplex) within VALHUDES, a framework established to assess the clinical performance of HPV assays on self-collected vaginal versus clinician-collected cervical samples. A total of 523 women, recruited at Belgian colposcopy clinics, provided vaginal self-samples using either the Evalyn Brush or Qvintip whereas gynaecologists subsequently collected cervical samples with a Cervex Brush. Both vaginal and cervical specimens were resuspended in 20&#x2009;mL of ThinPrep medium. Paired specimens from 499 women were tested with Allplex. The relative clinical sensitivity of vaginal self-samples collected with Evalyn Brush and Qvintip for high-grade cervical intraepithelial neoplasia (CIN2+) versus clinician-collected cervical samples was 0.96 (95% CI: 0.91-1.02) and 0.91 (95% CI: 0.82-1.01), respectively. Both self-sampling devices demonstrated higher specificity for < CIN2 compared to clinician-collected cervical samples, with relative specificity of 1.13 (95% CI: 1.02-1.25) on vaginal self-samples collected with Evalyn Brush and 1.09 (95% CI: 0.99-1.19) for those collected with Qvintip. Median Ct-values for overall hrHPV, HPV16, 31, 45, 52, 58 and human Beta-globin gene were significantly lower in clinician-collected cervical compared to vaginal self-samples. In conclusion, Allplex demonstrated comparable clinical sensitivity and higher specificity on vaginal self- versus clinician-collected cervical samples, without the need for cut-off optimization.