Folia histochemica et cytobiologica最新文献

Introduction: Oxidative damage-induced retinal pigment epithelial (RPE) cell apoptosis and optic nerve inflammation and demyelination are closely related to the pathogenesis of optic neuritis (ON). STAT1 has been found to be activated in the retina and optic nerve of ON rats. This study aimed to determine whether STAT1 depletion exerts neuroprotective effects against ON in both cellular and animal models.

Material and methods: ARPE-19 cells were stimulated by H₂O₂ to induce oxidative stress, followed by STAT1 and IFI30 silencing. CCK-8 and flow cytometry assays assessed ARPE-19 cell viability and apoptosis. RT-qPCR, Western blotting, DCFH-DA staining, and commercial kits detected the levels of STAT1, IFI30, apoptosis markers, and antioxidant/oxidative markers. CHIP and luciferase reporter assays validated the binding between STAT1 and IFI30 promoter. Female C57BL/6 mice were immunised with myelin oligodendrocyte glycoprotein (MOG) peptide (MOG35-55) to induce experimental autoimmune encephalomyelitis, an animal model of ON. Optic nerve inflammation, demyelination, axonal loss, and retinal ganglion cell (RGC) apoptosis in encephalomyelitis (EAE) mice after STAT1 knockdown were evaluated via haematoxylin and eosin, luxol fast blue, immunofluorescence, and Brn3a-TUNEL double staining.

Results: STAT1 silencing reversed the H₂O₂-induced increase of cell apoptosis and oxidative stress and the decrease in cell viability in ARPE-19 cells. STAT1 bound with the IFI30 promoter region and negatively regulated its expression. IFI30 knockdown antagonised the protection of STAT1 silencing against H₂O₂-induced oxidative stress and apoptosis in ARPE-19 cells. STAT1 depletion alleviated optic nerve inflammation, demyelination, axonal loss, and RGC apoptosis in EAE mice.

Conclusions: STAT1 silencing exhibits neuroprotective effects against ON by upregulating IFI30.

Introduction: O-GlcNAcylation is a post-translational modification in which a single N-Acetyl-D-Glucosamine (GlcNAc) molecule is added to Ser or Thr residues of proteins. The O-N-acetylglucosaminyl transferase (OGT) enzyme is responsible for adding GlcNAc to the target proteins and N-acetyl-β-D-glucosaminidase (OGA) that removes the GlcNAc residue. O-GlcNAcylation has been described in the pathophysiology of several diseases; however, little has been studied in dental tissue. The aim of the present work is to characterise the product of O-GlcNAcylation and its enzymes at the tissue level in the dental pulp, as well as its expression in dental pulp stem cells (DPSCs) both in situ and in vitro. This enables the recognition of the behaviour of O-GlcNAcylation in pulp tissue without pathology.

Material and methods: Pulp tissue was obtained from 10 healthy donors, and the expression of O-GlcNAc, OGT, and OGA was analysed using immunofluorescence with specific antibodies in different regions of the dental pulp. DPSCs were isolated, cultured, and identified with anti-STRO1 (antibody specific for human CD34+ cells, useful for DPSC identification). The expression of O-GlcNAc in DPSCs was confirmed in vitro through Western blot.

Results: Different regions of the dental pulp and DPSCs express O-GlcNAc and the enzymes OGT and OGA. O-GlcNAc and OGT expression was more prominent in the odontoblastic layer, cell-rich zone, and in the central core. OGA was distributed throughout the pulp tissue with lower immunoreactivity compared to OGT.

Conclusions: Our results suggest that O-GlcNAcylation may play a relevant role in human dental pulp homeostasis and in physiology of DPSCs.

Introduction: Bronchopulmonary dysplasia (BPD) is a common chronic respiratory disease in premature infants. Hyperoxia is the main pathogenic factor of BPD. Nintedanib is a small-molecule tyrosine kinase inhibitor that has been confirmed to affect several cellular processes in different diseases. The aim of this study was to explore the function of nintedanib in the treatment of BPD.

Material and methods: Newborn Sprague-Dawley rats (postnatal day 1) were used to establish an in vivo BPD model by hyperoxia induction. Nintedanib was intraperitoneally injected into rats. Haematoxylin and eosin (H&E) staining was applied to detect lung injury in BPD rats. Cell apoptosis was determined by TUNEL (TdT-mediated dUTP nick end labelling) assay. Western blotting was applied to detect level changes of inflammatory factors IL-1β (interleukin-1 beta), CXCL-1 (C-X-C motif chemokine ligand 1), MCP-1 (monocyte chemotactic protein-1), as well as the phosphorylation of IkB (NF kappa B inhibitor) and NF-kB (nuclear factor kappa-B) in lung samples.

Results: Hyperoxia resulted in lung injury in neonatal rats, while nintedanib treatment relieved the injury. Furthermore, nintedanib alleviated hyperoxia-induced apoptosis in rat lungs. It was further observed that an inflammatory response caused by hyperoxia in lung samples was attenuated by nintedanib administration. Additionally, nintedanib inactivated the NF-κB pathway in BPD rats.

Conclusions: Nintedanib alleviates hyperoxia-induced lung injury via targeting the NF-κB signalling pathway.

Introduction: . The adrenal glands are central regulators of endocrine homeostasis and stress adaptation. Despite decades of research, the molecular mechanisms governing adrenal zonation and cortical renewal remain incompletely understood.

Material and methods: . Here, the 10x Genomics Visium CytAssist platform was applied to generate a high-resolution atlas of the adult CD1 IGS male mouse adrenal gland. The study employed a range of analytical approaches, including unsupervised clustering, differential gene expression analysis, pathway and Gene Ontology enrichment, cell-type deconvolution, and pseudotime trajectory inference. These approaches collectively facilitated the comprehensive characterization of transcriptional signatures across the primary adrenal cortical zones and the medulla.

Results: . Herein transcriptional signatures of distinct compartments were identified, including zona glomerulosa/adrenal capsule (ZG/capsule), outer and inner zona fasciculata (oZF, iZF), medulla, connective tissue, and surrounding brown and white adipose depots. Deconvolution analysis further delineated contributions from stromal, immune, endothelial, and erythrocyte populations, highlighting the complexity of adrenal and periadrenal tissues. Pseudotime trajectory analysis validated the centripetal differentiation model of the adrenal cortex, demonstrating a continuum of transcriptional states from capsule and ZG/capsule through oZF to iZF, consistent with progressive acquisition of steroidogenic functions. Functional enrichment confirmed zone-specific pathways, including WNT and steroid biosynthesis in the cortex and neuroendocrine pathways in the medulla.

Conclusions: . Together, this study provides the first spatially resolved transcriptomic reference map of the adult male mouse adrenal gland, revealing the cellular and molecular framework underlying zonation, renewal, and inter-compartment interactions. This atlas constitutes a valuable resource for advancing the understanding of adrenal physiology and pathology.

Papillary thyroid carcinoma (PTC) constitutes the predominant subtype among thyroid malignancies. Despite its generally favorable prognosis, certain aggressive subtypes, along with recurrent and metastatic manifestations, substantially affect patient survival outcomes. Recent advancementsin the diagnostic and therapeutic strategies for PTC have ushered in a new era, characterized by the integration of molecular mechanisms and imaging-based evaluations. This review offers an integrated perspective of the clinicopathological features, molecular genetic characteristics, epigenetic regulation, and the contribution of the immune microenvironment to the aggressiveness of PTC. Primary inve stigation targets include BRAF/RAS/RET-related molecular mechanisms and the functional significance of non-coding RNAs [especially long non-coding RNAs (lncRNAs) and microRNAs (miRNAs)] in molecular regulation. Additionally, the impact of clinical factors such as age, sex, obesity, and comorbidity with Hashimoto's thyroiditis on the aggressiveness of PTC is thoroughly examined. Furthermore, this review systematically synthesizes the clinical advances in the early detection and risk assessment of aggressive PTC by emerging imaging modalities such as conventional ultrasound, interventional ultrasound, ultrasound elastography, contrast-enhanced ultrasound, and artificial intelligence-assisted analysis. Looking ahead, multidisciplinary collaborations integrating pathology, genomics, and imaging are anticipated to enhance the precise evaluation of PTC aggressiveness and facilitate the development of individualized treatment strategies. This review serves as a comprehensive reference for mechanistic exploration and clinical translation in the study of PTC aggressiveness, and provides guidance for the progression of precision medicine and management models for PTC patients.

Introduction: Sepsis, a life-threatening condition caused by a host response that goes out of control, leads to severe organ dysfunction. MicroRNA 939 (miRNA-939) plays a pivotal role in this process by post-transcriptionally regulating the mRNA expression of key enzymes in the NO/cGMP pathway. This pathway is crucial in the development of refractory hypotension and organ failure in sepsis. The aim of the study was to explore the effects of miRNA-939 on a caecum ligation puncture (CLP) rat model of sepsis in kidneys and lungs in order to elucidate its role in modulating the NO/cGMP pathway and related organ protection mechanisms.

Material and methods: To explore the effects of miRNA-939 on sepsis, three groups of rats were established: a sham surgery group, a sepsis model group, and an intervention group treated with LNA-antimiR targeting miRNA-939, which was administered via tail vein injection (the locked nucleic acid (LNA) group). miRNA-939 levels in serum, kidneys, and lungs were assessed using reverse transcription-polymerase chain reaction (RT-PCR). The protein levels of inducible nitric oxide synthase (iNOS) and soluble guanylate cyclase (sGC) were quantified using Western blotting. Serum nitric oxide (NO) levels were determined using the nitrate reductase method.

Results: Compared to the sham group, the rats in the sepsis model group exhibited significantly elevated levels of miRNA-939 in serum, with distinct peaks at 12, 24, and 36 h post-sepsis induction. Similarly, the protein levels of iNOS and sGC (sGCα1 and sGCβ1 subunits) in kidneys and lungs were markedly higher in the sepsis group. Serum NO levels were also significantly elevated in the sepsis group compared to the sham group. Importantly, inhibition of miRNA-939 in the LNA group led to a significant reduction in the tissue levels of iNOS, sGCα1, sGCβ1, and serum NO within the NO/cGMP pathway, alongside mitigating inflammatory damage to kidneys and lungs.

Conclusions: Inhibiting miRNA-939 significantly reduces the expression of iNOS, sGCα1, sGCβ1, and NO in the NO/cGMP pathway in CLP-exposed rats, thereby alleviating inflammatory organ damage in sepsis. These findings highlight the therapeutic potential of targeting miRNA-939 in the management of sepsis-induced organ dysfunction.

Introduction: This study investigated the therapeutic mechanisms of electroacupuncture (EA) in chronic unpredictable stress (CUS)-induced depression rat model. Since SIRT1 plays in oligodendrocyte differentiation and neuroprotection, we hypothesize that it may mediate the effect of electroacupuncture (EA) on myelin regeneration in depression.

Material and methods: Sixty adult male Sprague-Dawley rats were divided into control, CUS, EA + CUS and CUS + EA + SIRT1-specific inhibitor - EX527 (EX) groups. We established a CUS-induced depression rat model by subjecting rats to 4-week CUS paradigm. Four weeks post-modeling, EA treatment and intraperitoneal administration of EX527 were applied. Behavioral tests including open field test, forced swim test and sucrose preference test were performed to assess the depressive-like state. Immunohistochemistry and stereological analysis for quantification of oligodendroglial cell populations was performed. Immunohistochemical staining and transmission electron microscope were performed for evaluation of myelination. Western blot and qRT-PCR analyses were performed to detect the mRNA and protein expression of SIRT1 in the prefrontal cortex (PFC) of rats in each group.

Results: Four weeks of EA intervention significantly alleviated depressive-like behaviors in CUS rats, as evidenced by increased sucrose preference (P < 0.05), enhanced exploratory activity (P < 0.01), and reduced immobility time (P < 0.05) compared to the CUS group. Histopathological and ultrastructural analyses demonstrated that EA restored myelin integrity in the PFC, with myelin basic protein immunoreactivity significantly higher in the EA+CUS group than in untreated CUS rats (P < 0.05). Electroacupuncture promoted oligodendrocyte differentiation, reversing the chronic stress-induced reduction in CC1+/Olig2+/BrdU+ progenitor cells (31.5 ± 3.1% vs. 3.23 ± 1.4% in CUS + EA + EX group, P < 0.01). Mechanistically, EA upregulated SIRT1 mRNA and protein expression in the PFC (P < 0.05 vs. CUS), while pharmacological inhibition of Sirt1 with EX-527 abolished these effects, reducing Sirt1 mRNA and SIRT1 protein expression (P < 0.01, P < 0.01, respectively) and Olig2 expression (P < 0.05). EX-527 treatment also blocked EA-induced behavioral improvements and myelin regeneration, confirming the critical role of the Sirt1.

Conclusions: The findings indicate that EA ameliorates depression-like behavior by enhancing oligodendrocyte maturation and myelin repair via activating SIRT1 signaling. These findings provide novel mechanistic insights into non-pharmacological interventions for depression.

Introduction: Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy, encompassing distinct histological variants and a wide spectrum of clinical behaviors. Advances in molecular diagnostics have identified key genetic alterations - particularly BRAFV600E, RAS mutations, RET/PTC fusions, and TERT promoter mutations - that are strongly linked to tumor aggressiveness and prognosis. This study aimed to determine the prevalence of these alterations and evaluate their clinicopathological significance in a Saudi Arabian patient cohort.

Materials and methods: A retrospective analysis was conducted on 114 formalin-fixed paraffin-embedded (FFPE) PTC samples diagnosed between 2019 and 2023. Targeted next-generation sequencing (NGS) was used to detect BRAF, NRAS, KRAS, HRAS, RET/PTC fusions, and TERT promoter mutations. Immunohistochemistry (IHC) for BRAFV600E, TTF-1, CK19, HBME-1, and galectin-3 was performed using automated staining systems. Associations between genetic alterations and clinicopathological parameters - including tumor size, histological subtype, lymph node metastasis, and extrathyroidal extension - were analyzed statistically.

Results: BRAFV600E mutations were identified in 39.5% of cases and were significantly associated with larger tumor size (P = 0.01), extrathyroidal extension (P = 0.04), and lymph node metastasis (P = 0.03). RET/PTC fusions were detected in 15.8% of patients, predominantly younger individuals, and correlated with multifocal tumors and lymphovascular invasion. RAS mutations (19.3%) were more frequent in the follicular variant but showed no significant association with adverse features. TERT promoter mutations were present in 10.5% of cases, significantly correlating with older age and advanced tumor stage. IHC profiles demonstrated strong concordance with molecular findings.

Conclusions: In this Middle Eastern cohort of PTC patients, BRAFV600E and TERT promoter mutations were significantly associated with aggressive clinicopathological features, while RET/PTC fusions and RAS mutations demonstrated distinct demographic and histological distributions. The integration of NGS and IHC enhanced diagnostic accuracy and supports personalized risk stratification in PTC management.

Introduction: . Typhoid fever, a disease resulting from an infection with Salmonella Typhi (S. Typhi) remains widespread in economically disadvantaged regions, where it continues to be a critical public health concern. As the symptoms and signs are non-specific, they are difficult to diagnose directly based on the clinical picture. Therefore, laboratory examinations are essential for diagnosis.

Material and methods: . This research introduces a fast and equipment-independent approach for detecting S. Typhi by employing CRISPR/Cas12a-based technology. The optimized CRISPR/Cas12a system achieved a detection limit of 103 copies/μL of S. Typhi DNA per reaction, with the entire assay completed within 60 min.

Results: . Four clinical isolates cultured from patients with typhoid fever were collected and evaluated using our CRISPR/Cas12a-based detection system. The assay results demonstrated that all four samples were accurately identified as positive.

Conclusions: . We showed that the developed CRISPR/Cas12a-based detection method provides a promising alternative for the on-site and simple detection of S. Typhi.

Introduction: This study was aimed at exploring the effect of four units of polyethylene glycol (PEG4)-modified Ser-Val-Val-Tyr-Gly-Leu-Arg (SVVYGLR) peptide (SV peptide) on the proliferative potential and migrative capability of human gingival fibroblasts (HGFs) and the activation of adhesion-related proteins.

Material and methods: PEG4-SV peptide was synthesised using peptide solid phase synthesis. The proliferative response of HGFs to varying concentrations of PEG4-SV peptide was quantitatively evaluated using a Cell Counting Kit-8 (CCK-8) assay. The migratory capacity of HGFs in response to PEG4-SV peptide treatment was evaluated using an in vitro wound healing assay. The expression levels of adhesion-related genes, including collagen type I (COL-1), vinculin (VCL), focal adhesion kinase (FAK), and integrin β1(ITGB1), were quantitatively analysed using qRT-PCR. To assess the aforementioned adhesion-related proteins, immunofluorescence and Western blotting were performed.

Results: Primary HGFs were isolated through enzymatic digestion using dispase, and subsequently characterised by positive immunoreactivity for both vimentin and CD90 (Thy-1) markers. Compared to the control group, PEG4-SV at concentrations of 10, 20, and 40 μg/mL significantly promoted the proliferation of HGFs. The wound area in the SV group exhibited a significantly smaller size in the monolayer cell culture at 24 and 48 h. The expression of adhesion-related genes and proteins (collagen type I, vinculin, FAK and integrin β1), were significantly upregulated after treatment with 20 μg/mL PEG4-SV.

Conclusions: These results demonstrate that PEG4-SV peptide may have the ability to promote soft tissue healing around an implant surface and form tight soft tissue sealing on the transmucosal part of the implant.