Acta histochemica最新文献

{"title":"Non-specific background in immunoglobulin G staining can be effectively eliminated using heating or a catalase inhibitor","authors":"Kairan Yang ,&nbsp;Ting Xu ,&nbsp;Chengkang Lin ,&nbsp;Zuisu Yang ,&nbsp;Haiyan Lyu ,&nbsp;Falei Yuan","doi":"10.1016/j.acthis.2025.152269","DOIUrl":"10.1016/j.acthis.2025.152269","url":null,"abstract":"<div><div>Immunoglobulin G (IgG) staining is a widely used method for assessing blood-brain barrier (BBB) integrity. However, significant non-specific binding is often observed in many studies, which can interfere with the accurate interpretation of results. In this study, the horseradish peroxidase (HRP)-based polymer method and the streptavidin-biotin complex (SABC) method were used to perform IgG staining. The effects of hydrogen peroxide, heating, and a catalase inhibitor on reducing background staining were evaluated in brain sections from untreated mice and those subjected to middle cerebral artery occlusion (MCAO). The results showed that immunohistochemistry without hydrogen peroxide pretreatment still produced minimal background in paraffin-embedded sections. However, IgG staining with hydrogen peroxide pretreatment led to substantial background in vibratome sections. Compared to the SABC method, a mixture of the catalase inhibitor 3-amino-1,2,4-triazole and hydrogen peroxide reduced background staining by 35.4 % ± 5.7 % in the cortex of untreated mouse brains and by 36.9 % ± 1.8 % in the contralateral cortex of MCAO mice when using the polymer method. Additionally, heating at 75°C was sufficient to eliminate non-specific binding in brain sections from both untreated and MCAO mice. Hydrogen peroxide pretreatment alone was ineffective in removing background staining in brain sections from either untreated or MCAO mice. In summary, this study demonstrates that hydrogen peroxide pretreatment is effective in reducing background only when combined with a catalase inhibitor but is unnecessary when the tissue is heated. Heating is a simple and effective method for removing the IgG staining background when detecting BBB leakage.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 3","pages":"Article 152269"},"PeriodicalIF":2.3,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144084107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Retraction notice to “miR-103–3p attenuates liver injury with severe acute pancreatitis by inhibiting pyroptosis through miR-103-3p/NLRP1 axis” [Acta Histochem. 126 (2024) 152211]","authors":"Wenquan Zhang ,&nbsp;Min Du ,&nbsp;Yingjian Jiang ,&nbsp;Jiang Wang ,&nbsp;Yue Yu ,&nbsp;DianLiang Zhang","doi":"10.1016/j.acthis.2025.152258","DOIUrl":"10.1016/j.acthis.2025.152258","url":null,"abstract":"","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 3","pages":"Article 152258"},"PeriodicalIF":2.4,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144075339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantifying mechanical, proliferation, and migrational properties of bulk ovarian cancer cells and ovarian cancer stem cells","authors":"Crystal Brown ,&nbsp;Emily Stidham ,&nbsp;Elly Beck ,&nbsp;Ali Eickhoff ,&nbsp;Brianna Hill ,&nbsp;Kalyani Nair ,&nbsp;Craig Cady","doi":"10.1016/j.acthis.2025.152260","DOIUrl":"10.1016/j.acthis.2025.152260","url":null,"abstract":"<div><div>There are a limited number of studies analyzing ovarian cancer stem cell properties. The goal of this study was to analyze the mechanical and migrational properties of ovarian cancer stem cells with the well-researched bulk ovarian cancer cells. Through the use of atomic force microscopy (AFM), the mechanical properties of both cell types were gathered. In preparation for AFM analysis, an optimal fixation method was developed and performed on the cells. AFM analysis provided mechanical properties for both cell types, including cellular stiffness, maximum adhesion force, surface area, and mean surface roughness. Cell proliferation and transwell migration assays were assessed to determine the aggressiveness of both cancer cell types. A clear trend between both cell types was expected for the mechanical and potential aggression properties. The data from both analyses were used to create a baseline for migration, proliferation, and mechanical properties of ovarian cancer stem cells and bulk ovarian cancer cells. Further studies will assess if these properties are impacted by chemotherapy exposure.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152260"},"PeriodicalIF":2.3,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143931803","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “DRAM1 plays a tumor suppressor role in clear cell renal cell carcinoma through modulating Akt signaling” [Acta Histochem. 124 (2022) 151874]","authors":"Qingyan Feng,&nbsp;Meijuan Cheng,&nbsp;Jingjing Jin,&nbsp;Shenglei Zhang,&nbsp;Yaling Bai,&nbsp;Jinsheng Xu","doi":"10.1016/j.acthis.2025.152259","DOIUrl":"10.1016/j.acthis.2025.152259","url":null,"abstract":"","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152259"},"PeriodicalIF":2.3,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143958854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SNHG16 suppression enhances M2 macrophage polarization and inhibits VSMC migration in atherosclerosis","authors":"Bing Gao ,&nbsp;Maogong Xi ,&nbsp;Ying Cui ,&nbsp;Kai Wang ,&nbsp;Hui Zhang ,&nbsp;Yiyong Wang","doi":"10.1016/j.acthis.2025.152248","DOIUrl":"10.1016/j.acthis.2025.152248","url":null,"abstract":"<div><div>Atherosclerosis (AS) significantly impacts both cardiovascular and cerebrovascular health, making it an important area of research for potential therapeutic interventions. This study investigates the role of lncRNA SNHG16 in macrophage polarization and its effects on the progression of AS. We assessed the expression of SNHG16 in macrophages and vascular smooth muscle cells (VSMCs) treated with oxidized low-density lipoprotein (ox-LDL) using qPCR. Cell proliferation was evaluated via EdU assay and western blotting, while flow cytometry and immunofluorescence were employed to analyze the polarization of macrophages. Foam cell formation was examined using Oil Red O staining. In a co-culture system, VSMCs treated with ox-LDL were cultured alongside macrophages pretreated with sh-SNHG16, and VSMC viability, migration, and motility were assessed using CCK-8, migration, and scratch assays. The levels of inflammatory cytokines, including IL-6, IL-10, TGFβ, and TNFα, were quantified by ELISA. Our results show that SNHG16 expression is upregulated in ox-LDL-treated cells, which correlates with enhanced macrophage proliferation. Inhibition of SNHG16 promoted M1-to-M2 macrophage polarization, reducing foam cell formation and inflammation. Furthermore, SNHG16 knockdown limited VSMC viability and motility, while attenuating ox-LDL-induced inflammatory responses. In conclusion, suppression of SNHG16 favors M2 macrophage polarization and presents a potential therapeutic target for AS management.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152248"},"PeriodicalIF":2.3,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143906975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histological, histochemical, and morphometric analysis of epidermal Leydig cells and histochemical characterization of epidermal apical cells in juvenile and adult axolotls (Ambystoma mexicanum)","authors":"Omar Betancourt-León ,&nbsp;Verónica Rodríguez-Mata ,&nbsp;Antonieta Martínez-Guerrero ,&nbsp;Armando Pérez-Torres","doi":"10.1016/j.acthis.2025.152255","DOIUrl":"10.1016/j.acthis.2025.152255","url":null,"abstract":"<div><div><em>Ambystoma mexicanum</em>, also known as the axolotl, is a paedomorphic urodele. Metamorphosis can be induced experimentally, and the most significant changes occur in the skin. These include thinning of the epidermis, increased keratinization of the stratified squamous epithelium, and loss of Leydig cells (LCs). Similar epidermal changes are observed in other metamorphic urodeles. Epidermal cells are responsible for the secretory function of the skin in juvenile amphibians, whereas dermal glands perform this function in adults after metamorphosis. In the axolotl, this occurrence is still partially understood. The only recognized epidermal secretory cells in juvenile <em>A. mexicanum</em> are the LCs, whose specific secretion products have not yet been characterized from the histochemical standpoint. Additionally, the persistence of LCs in adulthood, when mucous and serous (granular-protein secretion) glands are abundant, remains a matter of debate. The present study aims to describe the morphological and histochemical changes in the epidermis of 10 cutaneous regions from juvenile (4 months old) and adult (24 and 48 months old) non-metamorphic <em>A. mexicanum</em>, with a particular focus on the amount and histochemical characteristics of LCs. Results indicate that the juvenile epidermis is a stratified cuboidal epithelium formed by three strata: basal, spinosum (containing the LCs), and apical. The most superficial layer contains cuboidal cells that lack the characteristics of a true stratum corneum. In adults, the stratum apical is also formed by squamous cells, suggesting a transition to a cornified and squamous layer as age increases. Histochemical methods demonstrated that LCs are most likely serous and not mucous cells. On the other hand, cuboidal cells of the juvenile apical stratum would be responsible for producing mucous secretion components. Morphometric analysis revealed a significant decrease in both LCs and the epidermal thickness in the 24-month-old adult axolotl compared to the juvenile. While LC count and epidermal thickness in the 48-month-old adult showed a slight increase compared to the 24-month-old adult, these differences were not statistically significant and far lower than those observed in the juvenile axolotl, which exhibited the highest number of LCs and a thicker epidermis. These natural axolotl epidermal changes indicate a gradual transition toward a morphology resembling metamorphic skin as age advances. The decreased number of LCs and the transition from cuboid cells to squamous cells in the stratum apical suggest that both cell types may naturally disappear entirely at some point during development.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152255"},"PeriodicalIF":2.3,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143864230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proposal for a simple and easy-to-implement protocol for three-dimensional tissue imaging that is compatible with observation using a confocal microscope","authors":"Takuto Matano ,&nbsp;Kiyotada Naitou ,&nbsp;Jannatul Ferdous ,&nbsp;Takahiko Shiina ,&nbsp;Mitsuya Shiraishi","doi":"10.1016/j.acthis.2025.152257","DOIUrl":"10.1016/j.acthis.2025.152257","url":null,"abstract":"<div><div>Tissue observation has traditionally been limited to obtaining two-dimensional information from thinly sliced tissues due to issues with light transmission and antibody penetration. In recent years, three-dimensional tissue observation methods combining tissue clearing and deep immunostaining methods have been reported. However, due to the significantly different procedures in these methods from conventional immunostaining methods and the requirement for an expensive and specialized light-sheet microscope for tissue observation, the widespread adoption of these methods has been limited. To promote the shift from the current two-dimensional tissue observation to three-dimensional tissue observation using a combination of tissue clearing and immunostaining, it is essential to establish a simple and easy-to-implement protocol that is compatible with observation using a confocal microscope, which is available in many facilities. In this study, we first examined the effects of tissue clearing and staining conditions of immunostaining with thin tissue slices. We showed that CUBIC-L enhances immunolabeling without diminishing the immunoreactivity of antigens. We also showed that high detergent concentrations enhance the intensity of immunoreactivity and that a two-step staining procedure is suitable for our proposed protocol. Based on the results, we propose a simple protocol that can be easily adapted from conventional methods and is compatible with confocal microscopes. The results of this study are expected to facilitate a shift from traditional methods to three-dimensional tissue observation techniques that combine tissue clearing and immunostaining, contributing to the broader adoption of three-dimensional tissue observation.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152257"},"PeriodicalIF":2.3,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression characteristics and biological functions of CGB5 gene in gastric cancer","authors":"Fuping Gao ,&nbsp;Xiaohua Zhou ,&nbsp;Jin Wei ,&nbsp;Qiong Sun ,&nbsp;Jiapeng Wang ,&nbsp;Qing Li","doi":"10.1016/j.acthis.2025.152254","DOIUrl":"10.1016/j.acthis.2025.152254","url":null,"abstract":"<div><div>Objective: The chorionic gonadotropin (CG) subunit beta 5 (CGB5) gene is a member of the glycoprotein hormone β chain family, encoding the β5 subunit of CG, which has been shown to promote tumorigenesis and induce proliferation in various types of cancer including gastric cancer (GC). However, the mechanistic role of CGB5 in GC has not been fully elucidated. Therefore, this study investigated relevant genes that regulate GC through bioinformatics analysis. Methods: Immunohistochemistry, immunofluorescence, and western blot (WB) detection methods were appropriately used to evaluate the expression pattern and clinical significance of CGB5 in 100 Chinese GC patients that were recruited from the Gaochun People's Hospital. The effect of small interfering ribonucleic acid (siRNA) on apoptosis, migration, and invasion of GC cells was investigated in vitro. Three-dimensional tumor spheres of these two types of GC cells (NCI-N87 cells and MKN45 cells) were constructed before investigation of the Calcein acetoxymethyl ester (AM)/ Propidium iodide (PI) staining, flow cytometric apoptosis, and apoptotic-related protein content of the tumor spheres after siRNA inhibition of CGB5 expression. Results: It was observed that compared with adjacent normal gastric tissue, expression of CGB5 was significantly upregulated in GC tissue. The siRNA inhibited CGB5 expression in two GC cell lines (NCI-N87 cells and MKN45 cells). Also, it was discovered that CGB5 highly correlated with microsatellite instability (MSI) and immune cell activity in GC, thus revealing the greater research value of CGB5 gene. More importantly, CGB5 siRNA could inhibit invasion and migration of tumor cells, induce apoptosis of GC cells and GC tumor spheres, as well as the mechanism relating to regulation of apoptosis associated gene expression. Overall, the findings suggest that CGB5 may play a crucial role in the development of GC carcinogenesis. Thus, this research may contribute to design of potential drug targets for treatment of GC.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152254"},"PeriodicalIF":2.3,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143852056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery of nuclear cavities in Epstein-Barr virus-infected cells","authors":"Shinji Shimada ,&nbsp;Hideya Kawasaki ,&nbsp;Hiroto Katoh ,&nbsp;Shumpei Ishikawa","doi":"10.1016/j.acthis.2025.152253","DOIUrl":"10.1016/j.acthis.2025.152253","url":null,"abstract":"<div><div>Approximately 90 % of humans are infected with the Epstein-Barr virus (EBV); however, most do not develop neoplastic lesions. Despite various investigations, the underlying reasons remain largely unknown. Therefore, we aimed to address this question through morphological observations to identify the ultrastructural alterations occurring in EBV-infected cells. EBV-positive cells from legacy lymph node specimens obtained from patients with HIV and modern fresh specimens of aberrantly proliferating human EBV-positive lymphocytes in a patient-derived xenograft (PDX) of immunodeficient mouse were examined. By utilizing a special technique that allowed us to observe exactly the same specimen using both optical and electron microscopy, we were able to detect a peculiar phenomenon in EBV-infected cells. EBV-infected lymphocytes occasionally exhibited nuclear cavities, a finding that was confirmed in multiple specimens from both patients with HIV and the PDX model. It was suggested that EBV-infected cells may activate cell death pathways, based on the protein expression patterns of p53 and FAS. Taken together, these results indicated that nuclear cavity formation appeared to be a characteristic morphological alteration associated with EBV infection. Further research could clarify the potential relationship between nuclear cavities and cellular biology in EBV-infected cells, possibly shedding light on the mechanisms that prevent EBV-infected cells from progressing to tumor formation.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152253"},"PeriodicalIF":2.3,"publicationDate":"2025-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of collagen I and collagen III immunohistochemistry with Herovici staining in various rabbit organs","authors":"Gabriella Meier Bürgisser,&nbsp;Pietro Giovanoli,&nbsp;Maurizio Calcagni,&nbsp;Johanna Buschmann","doi":"10.1016/j.acthis.2025.152256","DOIUrl":"10.1016/j.acthis.2025.152256","url":null,"abstract":"<div><div>Collagen I and III distribution is not only crucial to assess the status of healing wounds, but also to characterise healthy connective tissue and pathological extracellular matrix composition. In this technical note, we have therefore compared the dual-coloured Herovici staining, indicating pink collagen I and blue collagen III in serial sections with immunohistochemistry (IHC) labellings for collagen I and III, respectively. Furthermore, we used, chromogenic DAB for IHC labelling. Seven different organs of a healthy New Zealand white rabbit were collected for this purpose, including kidney, liver, tonsil, tongue, duodenum, heart, and brain, respectively. A dual-coloured staining like Herovici turned out to be as good as two single-colour labellings utilising IHC. In some cases, co-localisation and extent of collagen I and III expression could be qualitatively visualised better using Herovici, with gradients of blue-violet-pink, than by mere comparison of labelling intensities side by side in two different sections, although taken at the same place as serial sections. Nevertheless, a quantitative analysis of the Collagen I-to-III ratio revealed no significant differences between these two approaches to assess the extracellular matrix composition. From these comparisons, we conclude that a Herovici staining is recommended as a valuable alternative staining to collagen I and III IHC; and it may act as a fast and cheap preliminary staining method. These findings encourage researchers focusing on ECM composition of the experimental rabbit tissue to use Herovici staining to determine the ratio of the extracellular collagen I and III expression.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 2","pages":"Article 152256"},"PeriodicalIF":2.3,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143843744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}