European Journal of Histochemistry最新文献

Newborns' intestinal adhesions have been reported in 4.7% infants who underwent a laparotomy, but adhesions can also appear idiopathically. Etiology and pathogenesis of adhesions is still to be determined, but evidence shows relation to inflammation, formation of fibrin bands, hypoxia and tissue remodelation. Multiple candidate genes have been associated with adhesion development. The aim of this study was to evaluate the appearance of Sonic Hedgehog (SHH), Indian Hedgehog (IHH), Forkhead-box F1 (FOXF1), caudal type homeobox 1 (CDX1), HCLS1-associated protein X-1 (HAX-1), GATA Binding Protein 4 (GATA4) and Granzyme-B (GZMB) proteins in infant adhesions and to describe possible interfactorial correlations. Adhesion affected tissue samples were collected from 14 patients under one year of age that underwent abdominal surgery to treat partial or complete intestinal obstruction. The control group consisted of 6 individuals that had surgical repairment of inguinal hernia. Routine staining and immunohistochemistry were performed. Immunopositive fibroblasts, macrophages, endotheliocytes, smooth muscle myocytes of blood vessel wall and mesotheliocytes were investigated. The relative distribution of all factors was evaluated by the semiquantitative counting method. Statistical analysis was done using non-parametric tests and correlations were calculated based on Spearman's correlation analysis. A statistically significant decrease was observed for SHH, IHH, FOXF1, GATA4 and partially for GZMB in the adhesion group. There were also decreased HAX-1 and CDX1 immunopositive structures in the adhesion group, however, without any statistical significance. SHH, IHH, FOXF1, GATA4 and GZMB might have a role in adhesion development among infant patients which could suggest a dysregulation of cellular events. Abundance of correlations between the gene protein appearances in different structures indicate the affected blood vessels, fibroblasts and macrophages, however, mesothelium seems not to be the key driver in the morphopathogenesis of adhesion development.

Morphological analysis of neuronal processes and their networks is a key aspect of neuroscience, with relevance from basic research to clinical practice due to the central role of neuronal development and plasticity in many neurological disorders. More than a century after its introduction, Golgi staining, a technique based on the random precipitation of metallic deposits in different neuronal subtypes, remains a highly valuable method for investigating the cellular morphology of neurons in the nervous system. Despite the wide range of protocols developed over the years, several limitations of the technique remain a matter of discussion. Among these is the need to extend sample preservation during the interval between staining and sectioning procedures without compromising the quality of the histochemical labeling. By adopting a specific processing method, the present study demonstrates that it is possible to embed murine nervous tissue following Golgi staining and to preserve the samples for extended periods prior to sectioning, while maintaining well-preserved and clearly detectable histochemical labeling across different regions and neurons of the mouse central nervous system.

Hypoxia is a key driver of glioblastoma (GBM) progression. Serine/arginine-rich splicing factor 3 (SRSF3) is associated with the malignant progression of GBM, but its role in the hypoxic microenvironment of GBM remains unclear. This study aimed to explore the regulatory role and molecular mechanisms of SRSF3 in hypoxia adaptation in GBM. The expression of SRSF3 in normal astrocytes and GBM cells was detected. The effects of knockdown or overexpression of SRSF3 combined with hypoxia treatment on malignant phenotypes and hypoxia stress adaptation in GBM cells were evaluated. Cell viability, colony formation, migration, invasion, and cell death assays were performed to assess phenotypic changes. Mechanisms were investigated using mRFP-GFP-LC3, autophagy, and unfolded protein response (UPR)-related molecular detection. SRSF3 was highly expressed in GBM cells. Knockdown of SRSF3 inhibited cell viability, migration, invasion, and colony formation, whereas overexpression of SRSF3 promoted malignant behaviors. Further studies revealed that hypoxia induction significantly increased the expression levels of GRP78, CHOP, ATF4, LC3-II/I, and p62; upregulated the GFP/mRFP ratio; and increased cleaved-caspase3 expression, promoting cell death. Mechanistic studies revealed that SRSF3 overexpression promoted XBP1s formation, alleviated hypoxia-induced autophagic flux blockage, and reduced cell death. The IRE1 RNase inhibitor 4μ8C weakened the SRSF3-mediated promotion of XBP1s generation. SRSF3 enhances adaptive UPR output by promoting IRE1-dependent XBP1 splicing, thereby maintaining autophagic flux and promoting GBM cell survival under hypoxic conditions.

G protein-coupled estrogen receptor 1 (GPER1) has extensively verified as a tumor regulator in various types of cancers. However, its role in esophageal cancer (EC) remains largely unclear. In this study, the expression and prognostic prediction value of GPER1 in EC was analyzed by using TCGA database and was verified in EC cells and fresh tissues. The results showed that GPER1 is decreased in EC cells and tissues, and lower GPER1 expression is associated with poor overall survival of EC patients. CCK-8 assay and flow apoptosis cytometry were applied to measure the ability of proliferation and apoptosis of EC cells with or without GPER1 overexpression. The levels of reactive oxygen species (ROS) and Fe2+ were determined by flow cytometry. Elisa and Western blotting were employed to measure the markers of ferroptosis and cyclic adenosine monophosphate (cAMP) pathway. The results of in vitro experiments indicated that overexpression of GPER1 caused decreased proliferation, increased cell apoptosis, ROS generation, Fe2+ content and acyl-CoA synthetase long-chain family member 4 (ACSL4) expression, while decreased glutathione peroxidase 4 (GPX4) expression. Notably, the cAMP/PKA inhibitor H89 significantly reversed the ferroptotic effects induced by GPER1, indicating the essential role of the cAMP pathway in this process. The weight and volumes of tumors were measured and Ki-67 and H&E staining were conducted to analyze the effect of GPER1 in vivo. The results of in vivo experiments indicated that overexpression of GPER1 resulted in restricted tumor growth, reduced Ki-67 expression and increased cell death. In conclusion, the expression of GPER1 is reduced in EC. Overexpression of GPER1 enhances ferroptosis in EC, primarily through activation of the cAMP signaling pathway.

Ozone (O3) is an oxidizing natural gas widely applied as adjunctive therapeutic treatment for a variety of pathological conditions. Currently, O3-based therapies rely on the low-dose concept i.e., the administration of low O3 concentrations able to induce a mild oxidative stress stimulating antioxidant and anti-inflammatory response without causing cell damage. In addition, low O3 concentrations are thought to activate cellular and molecular mechanisms responsible for analgesic and regenerative effects. Due to these properties, in the last decade interest has arisen in the fields of orthopedics and regenerative medicine on the potential of O3 to counteract joint diseases involving cartilage degeneration. In this pilot study, we have explored the anti-degenerative potential of O3 on knee articular cartilage explanted from a healthy adult rabbit and maintained in vitro. Light and transmission electron microscopy were used to monitor chondrocyte and extracellular matrix features of cartilage samples undergoing O3 treatment every three days for two weeks. Results demonstrated that low O3 concentrations act on chondrocytes and the molecular components of the extracellular matrix of articular cartilage explants, significantly improving their preservation under in vitro conditions, likely by promoting both protective and pro-regenerative pathways. This opens promising perspectives for further investigations on the therapeutic potential of O3 for the treatment of cartilage degeneration not only as painkilling and anti-inflammatory agent but also as a cartilage regenerative agent.

Sepsis remains a major cause of morbidity and mortality worldwide, yet its prolonged pathophysiological consequences are poorly understood. Here, we employed a murine cecal ligation and puncture (CLP) model to investigate the prolonged impact of sepsis on survival, systemic inflammation, and organ pathology. Adult male C57BL/6 mice underwent CLP or sham surgery and were monitored for 28 days. Survival was recorded daily, while serial assessments of hematology, serum biochemistry, bacterial load, and cytokine levels were performed. Tissue immunofluorescence was used to characterize myeloid-derived suppressor cells (MDSCs), which are potent immunosuppressive cells that inhibit both adaptive and innate immune responses in sepsis, contributing to sepsis-induced immunosuppression. Histopathological analyses were conducted to evaluate structural changes in major organs. CLP mice displayed markedly reduced long-term survival compared with sham controls. Hematological profiling revealed persistent leukocytosis and an inflammatory response, while serum analyses showed sustained elevations in in bilirubin, creatinine, and blood urea nitrogen, reflecting hepatic and renal injury. Bacterial cultures confirmed systemic microbial persistence, and cytokine measurements indicated ongoing inflammatory activity. Tissue immunofluorescence demonstrated the infiltration of MDSCs across multiple organs, consistent with post-sepsis immunosuppression. Histopathological examination revealed widespread, chronic injury in the lungs, liver, kidneys, and spleen, including inflammatory infiltration, tissue degeneration, and architectural disruption. In conclusion, sepsis induces not only acute systemic inflammation but also enduring immune dysregulation and progressive organ damage. These findings highlight the CLP model as a robust platform for studying post-sepsis sequelae and underscore the need for therapeutic strategies that target long-term organ protection and immune restoration.

Inflammatory bowel disease (IBD) including ulcerative colitis (UC) and Crohn's disease has become a global disease in the 21st century, with increasing incidence rates in almost every industrialized country. Previous studies have suggested that the traditional Chinese medicine herb, cryptotanshinone (CTN), a major liposoluble extract of Salvia miltiorrhiza, alleviates the symptoms of experimental colitis in vitro and in vivo. However, the mechanisms underlying the protective effects of CTN against IBD remain exclusive. The present study found that CTN reversed lipopolysaccharide-induced inflammation in human colon epithelial cells (HIEC-6) by inhibiting the NF-κB pathway. In addition, CTN alleviated dextran sulfate sodium (DSS)-induced inflammatory bowel disease in mice by regulating the balance of TH17/Treg cells. CTN also exerted its role by inhibiting the polarization of M1 macrophages in mice with DSS-induced colitis. Of note, the effects of CTN on these immune cells may be mediated via changes in the levels of TNF-α and IL-6 directly in mice. Taken together, these findings may provide new insight regarding the therapeutic potential of CTN for UC.

We identified vagal innervation in the pharyngeal tooth and jawbones of Nile tilapia through macroscopic observations and immunohistochemistry. We also revealed the apposition of the nerve and osteoclasts in the pharyngeal jaw, suggesting the possibility of neuronal regulation for bone remodeling. However, the central projection from the vagal nerve, which innervates the pharyngeal jaws, remains unknown. To determine the projection of the vagus nerve in the brain, we applied carbocyanine dye (DiI) into the vagus nerve, revealing DiI-labeled neurons in the caudal vagal ganglion. The labeled fibers of the neurons were then traced to the vagal lobe, revealing that they branched and ran dorsally before terminating in a band-like pattern. Meanwhile, the labeled fibers running ventral to the vagal lobe were directed toward the dorsal motor nucleus of the vagus and did not have a definite terminal structure. The vagus nerve innervates the pharyngeal jaw, mainly projects to the vagal lobe, where it receives gustatory information. Pharyngeal tooth-derived sensory information might occur during occlusion and be processed precisely for determining the regurgitation and swallowing of prey.

Cold shock domain (CSD) proteins, such as YB-1, play a crucial role in the regulation of transcription, mRNA stability, and translation. Consequently, YB-1 is implicated in processes such as cell differentiation, oncogenesis and oxidative stress response. The development of the eye is a complex process that involves the differentiation of numerous highly specialized cell types. We hypothesized that YB-1 is involved in both eye development and stress defense mechanisms. As an initial step, we investigated the expression of YB-1 during the embryology of the mouse eye. YB-1 mRNA could be detected by RT-PCR and sequencing the PCR product in retinal tissue of adult mice. To elucidate the expression pattern of YB-1 protein during mouse eye development, we analyzed its expression in the developing mouse eye at embryonic day 13 (E13), E15, E18 and postnatal day 14 (P14) using immunohistochemistry. Expression of the YB-1 protein was detected in all retinal cells, as well as in the corneal and lens epithelial cells, throughout all stages of eye development examined. These findings suggest that YB-1 could have a significant role in the eye, potentially related to development and differentiation.

Metabolism-associated fatty liver disease (MAFLD) is a liver disease characterized by hepatic steatosis and excessive accumulation of lipids, with a high global incidence, especially in populations with obesity, diabetes and metabolic syndrome (MetS). As an important B vitamin, folate (FA) is stored mainly in the liver where it regulates oxidative stress, chronic inflammation and lipid metabolism. However, its regulatory role and mechanism of action in MAFLD are still poorly understood. Therefore, this study was conducted to investigate the regulatory effect of FA on MAFLD. The MAFLD rat model was induced by a high-fat diet (HFD), and HepG2 cells were treated with 0.3 mM palmitic acid (PA) for 24 h to establish a cell model. The expression of relevant genes and proteins was detected by RT-qPCR and Western blotting. Injury to HepG2 cells and rat liver tissues was evaluated via hematoxylin and eosin staining, Oil red O staining, ELISA and CCK-8 assay. FA treatment inhibited body weight gain in rats and reduced the levels of liver injury indicators (aspartate and alanine aminotransferase, and Alkaline phosphatase), blood lipids (total cholesterol, triglycerides and free fatty acids) and inflammatory cytokines (TNF-α, IL-6, and IL-1β), reducing lipid accumulation and pathological damage in the liver and ultimately alleviating the progression of MAFLD. Moreover, FA treatment promoted the expression of the autophagy-related protein LC3 II/I, inhibited the expression of p62, and increased the formation of autophagosomes, thereby alleviating PA-induced damage to HepG2 cells. Furthermore, NRF2 expression is downregulated in MAFLD and can be upregulated by FA treatment. Further examination revealed that knocking down NRF2 could partially attenuate the inhibitory effect of FA on PA-induced HepG2 cell injury. In conclusion, FA activates autophagy by promoting the expression of NRF2, thereby alleviating the development of MAFLD.