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IntroductionMethodsResultsDiscussionIn this study, our aim was to investigate the protective effects of myricetin (single dose–100 mg/kg) on CLP-induced rat sepsis model by analyzing some immune mechanisms including inflammation and oxidative stress by different techniques such as Immunohistochemistry, ELISA, tissue biochemistry and Western Blotting.Twenty-eight Wistar albino rats were divided into 4 groups. The pro-inflammatory and anti-inflammatory cytokine levels were measured by ELISA technique. CD68 and Nuclear-Factor-Kappa-B (NF-κB) positivity rates were detected by IHC. Some of oxidative stress parameters were measured by tissue biochemistry, while Toll-lke Receptor-4 (TLR4) expression others were detected by Western blot technique.Sepsis caused a significant increase in all pro-inflammatory cytokine and oxidant levels. Also, it led to an increase in the positivity of CD68 and NF-κB markers as well as the expression levels of TNF-alpha, IL-1-beta, TLR4, Keap-1. However, single dose myricetin application normalized pro-inflammatory cytokine levels, increased anti-oxidant and anti-inflammatory cytokine levels, decreased positivity of CD68 and NF-κB and increased NRF2 and HO-1 expressions.As a conclusion, the beneficial effect of myricetin on lung injury also involved inhibition of TLR4/NF-κB pathway, suppression of proinflammatory cytokines and induction of anti-inflammatory cytokine production, regulation of oxidant and anti-oxidant system parameters, and activating the NRF2/Keap1/HO-1 pathway. [ABSTRACT FROM AUTHOR]

This study aims to investigate the effect of estrogen hindrance, i.e., menopause in women for instance with rheumatoid arthritis on the brain hippocampal region by using collagen-induced arthritis (CIA) female rat model (RA). CIA was induced in female rats by injecting bovine type II collagen and incomplete Freund's adjuvant. Estrogen receptor antagonist, fulvestrant (Ful), was given to RA rats to create estrogen hindrance. Control (C) and RA rats were injected with saline and DMSO, respectively, while RA + Ful rats received a 7-day fulvestrant injection. Following experiment completion, rats were sacrificed, and brains were harvested. Brains were stained with H&E and cresyl violet staining and morphological changes in the hippocampus were identified. Additionally, oxidative stress, inflammatory, and apoptosis markers' levels in the hippocampus were analyzed by qPCR, ELISA, and immunohistochemistry techniques. RA + Ful rats showed neuronal atrophy and reduced neurogenesis in the hippocampal regions. NOX4, NF-κB, IL-1β, IL-6, TNF-α, IKK-β, and Bax protein expression levels in the hippocampus were increased, whereas hippocampal Bcl-2, caspase-3, caspase-9, and IGF-1R protein expression levels were decreased. Furthermore, RA + Ful rats had lower levels of antioxidants PON-1 and catalase in the hippocampal regions. The changes in these molecular markers were statistically significant when compared to RA rats without Ful treatment (p < 0.05). Estrogen hindrance exaggerated oxidative stress, inflammation, and apoptosis which resulted in neuronal degeneration in the hippocampal regions in rheumatoid arthritis. [ABSTRACT FROM AUTHOR]

The article discusses the use of a five-plex Hepatic Oncochip to study liver cancer, focusing on the epithelial-mesenchymal transition (EMT) and the biomarker BAP31. The study aims to explore tumor heterogeneity in hepatocellular carcinoma (HCC) and the relationship between EMT and BAP31. By utilizing multiplex immunohistochemistry (mIHC) technology, the researchers analyzed 138 tissue samples, providing insights into the molecular heterogeneity of HCC. The findings suggest a strong correlation between elevated BAP31 expression and poor prognosis in liver cancer, challenging traditional views on EMT in hepatocellular carcinoma. [Extracted from the article]

Carnivorous plants have fascinated botanists and ecologists with their various unusual adaptations in organ structure, physiology, and complex interactions with other organisms since the time of Charles Darwin. Species of the genus Utricularia (bladderworts, family Lentibulariaceae) are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. In the traps, there are glandular trichomes called quadrifids, which produce digestive enzymes and absorb the products of prey digestion. These quadrifids are unique due to their highly complex glandular cell structure; hence, they are an excellent model for studying the cell wall and its specialization. The main aim of the study was to investigate the presence and distribution of homogalacturonans (HGs) and hemicelluloses in the cell walls of trichome cells and especially in cell wall ingrowths in the quadrifid cells. The following antibodies were used against the wall components: anti-HGs (homogalacturonans) —JIM5 (low methylesterified HGs), JIM7 (highly esterified HGs), LM19 (low methylesterified HGs), CCRC-M38 (a fully de-esterified HG), LM5 (galactan); anti-hemicelluloses—LM25 (galactoxyloglucan; XXLLG, XXLG, XXXG modules of xyloglucans), LM15 (xyloglucan), CCRC-M138 (xylan), LM11 (heteroxylan); and anti-mannans: LM20 (heteromannan) and LM22 (heteromannan). The localization of the examined compounds was determined using immunohistochemistry techniques and immunogold labeling. In quadrifid cells, we found differences in the presence of the epitope detected by the LM5 antibody in the cell walls. In addition, cell wall ingrowths represented distinct microdomains of the cell wall in terms of the occurrence of wall components (they were methylesterified and demethylesterified homogalacturonan-poor). Hemicelluloses (galactoxyloglucan and xyloglucan) and arabinogalactans co-occur in cell wall ingrowths. Also, a part of the cell wall of the pedestal cell, which forms a Casparian strip, represented a distinct microdomain. We did not detect epitopes recognized by LM11, LM20 and LM22 antibodies. Our research shows that several cell wall microdomains occur in the cell walls of quadrifid cells. They differ depending on the presence and distribution of low methylesterified HGs, highly esterified HGs, fully de-esterified HGs, galactan (the epitope detected by the LM5 antibody), xyloglucan, galactoxyloglucan, and xylan (the epitope detected by the CCRC-M138 antibody). [ABSTRACT FROM AUTHOR]

Background: Abnormal expression of Zinc finger (ZNF) genes is commonly observed in osteosarcoma (OS), the most prevalent malignant bone tumor in children and teenagers. This project focused on the role of ZNF560 in the progress of OS. Methods: The published datasets including TCGA-SARC and GSE99671 was utilized to screen out the abnormal expression of ZNF560 and associated gene patterns in sarcoma and OS tissues. Prognosis value of ZNF560 was identified in TCGA-SARC and OS cohorts. In order to manipulate ZNF560 expression in HOS and MG63 osteosarcoma (OS) cells, genetic strategies such as shRNA constructs were utilized. The expression patterns of ZNF560 were analyzed through techniques such as immunohistochemistry, Western blotting, and qRT-PCR. Results: By analyzing data from both the GEO and the Cancer Genome Atlas (TCGA) databases, increased expression of ZNF560 in OS tissues was verified, which was significantly associated with poorer outcomes in osteosarcoma patients both in TCGA-SARC and our own OS cohorts. Additionally, downregulation of ZNF560 resulted in decreased cell viability, fewer colonies, and induced apoptosis of osteosarcoma cells. Moreover, ZNF560 was found to be essential for migration of human osteosarcoma HOS and MG63 cells. Conclusion: Collectively, these findings suggest that ZNF560 has the potential to serve as a predictive biomarker for osteosarcoma. [ABSTRACT FROM AUTHOR]

Background: Lung cancer is a globally pervasive and deadly disease, claiming more than 1 million lives annually. Therefore, the identification of mutations in crucial cancer-related genes is paramount for guiding optimal chemotherapy strategies. The distribution of EGFR, KRAS, ALK, and ROS1 mutations varies across diverse ethnic populations. Nonetheless, there is limited data available on the prevalence of these mutations and their correlation with PD-L1 expression among Iranian lung cancer patients. Aim: This study involved an analysis of EGFR, KRAS, ALK, and ROS1 gene mutations in lung cancer patients, followed by an assessment of the correlation between PD-L1 expression and clinicopathological variables. Methods: Mutational profiling was conducted by examining EGFR (exons 18–21) and KRAS (exon 2) through pyrosequencing. Detection of ALK and ROS1 rearrangements, alongside PD-L1 expression, was carried out using immunohistochemistry techniques. Results: EGFR mutations were identified in 23.4% of cases, exhibiting a notably higher occurrence in females (p = 0.001). KRAS mutations were present in 7.1% of cases, with no significant association found between KRAS mutations and sex (p = 0.229). ALK rearrangements were found in 4.9% of cases, while ROS1 rearrangements were present in 0.6% of patients. The overall prevalence of PD-L1 protein expression was 36.85%. Notably, PD-L1 expression was detected in 24.8% of cases with EGFR mutations, 20% of cases with KRAS mutations, 64.7% of cases with ALK rearrangements, and in 100% of cases with ROS1 rearrangements. Conclusion: Although no correlation was found between PD-L1 expression and EGFR, KRAS mutations, and ROS1 rearrangements, a noteworthy association was identified between ALK rearrangements and elevated PD-L1 expression. [ABSTRACT FROM AUTHOR]

Objective: Heavy metals and environmental pollutants, such as cadmium chloride, congenital disorders, and certain diseases, can lead to infertility in men. In this study, the effects of lupeol (an active pentacyclic triterpenoid with antioxidant properties) on testicular injuries induced by cadmium chloride were investigated in male rats. Materials and Methods: Lupeol was obtained from Sigma-Aldrich, and the experiment included 40 male Wistar rats divided into 8 groups (healthy control, healthy rats treated with 100, 200, and 400 mg/kg of lupeol, cadmium chloride, and three groups that received cadmium chloride and were treated with 100, 200, and 400 mg/kg of lupeol). After oral treatment, rats were anesthetized, and blood and testicular tissue sampling was done. Subsequent analysis of oxidative stress enzymes, such as malondialdehyde (MDA) and superoxide dismutase (SOD), testosterone, sperm motility, and Aquaporin 9 (AQ9) levels was performed using ELISA, histopathology, and immunohistochemistry techniques, respectively. Immunohistochemistry staining was done for expression of Aquaporin 9 in seminiferous tubules. Results: The results showed that compared to the healthy control, cadmium chloride caused a significant decrease in SOD, testosterone, sperm motility, and sperm vitality, with severe destruction of spermatogenic tubes and a significant increase in MDA and AQ9 rate. Rats which were treated with cadmium chloride+400 mg/kg of lupeol showed a significant increase in SOD, testosterone, histopathology, sperm motility, and sperm vitality rate, and significant organization of spermatogenic tubes in testis tissue. There was also a decrease in MDA and AQ9 of rats that received high-dose lupeol. (p<0.001) Conclusion: This study suggests that lupeol has a high potential for improving male reproduction and antioxidants in rats exposed to oxidative stress. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a neurodegenerative condition characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) of the brain, manifesting itself with both motor and non-motor symptoms. A critical element of this pathology is neuroinflammation, which triggers a harmful neurotoxic cycle, exacerbating cell death within the central nervous system. AD-16 (also known as GIBH-130) is a recently identified compound capable of reducing the expression of pro-inflammatory cytokines while increasing the expression of anti-inflammatory cytokines in Alzheimer's disease models. Here, for the first time, we sought to comprehend the potential impact of orally administered AD-16 in mitigating neurodegeneration and subsequent disease progression in PD. To accomplish this, 6- hydroxydopamine (6-OHDA) unilateral striatal injections were employed to induce a PD model in male C57BL/6 mice. Cylinder and apomorphine-induced rotation behavior tests were conducted to assess motor behavior and validate the PD model 3 days after the injection. AD-16 was administered via gavage daily between days 3 and 9 after surgery. On the last day of treatment, motor tests were performed again. All animals were euthanized on day 10 and immunohistochemistry techniques were performed to detect tyrosine hydroxylase (TH) and Iba-1 and thus label dopaminergic neurons and microglia in the SNc and striatum (CPu). These same regions were collected for ELISA assays to assess different cytokine concentrations. Our results revealed an enhancement in the motor function of the AD-16-treated animals, as well as reduced nigrostriatal neurodegeneration. In addition, AD-16 reduced the increase in microglia density and prevented the changes in its morphology observed in the PD animal models. Furthermore, AD-16 was able to avoid the increase of pro-inflammatory cytokines levels that were present in 6-OHDA-injected animals who received vehicle. Consequently, AD-16 emerges as a compound with significant potential for negative modulation of neurodegeneration and neuroinflammation suppression in the 6-OHDA animal model of Parkinson's disease. [ABSTRACT FROM AUTHOR]

The Utricularia (bladderworts) species are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. On the outer surfaces of the trap, there are dome-shaped glands (capitate trichomes). Each such trichome consists of a basal cell, a pedestal cell, and a terminal cell. During the maturation of these external glands, there are changes in the cell wall of the terminal cell of the gland (deposited layers of secondary wall material). Thus, due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls. The main aim of this study was to check whether different cell wall layers in terminal gland cells have a different composition in the case of homogalacturonans (low-methylesterified HGs, fully de-esterified HGs, and galactan) and hemicelluloses (galactoxyloglucan, xyloglucan, and xylan). The antibodies were used against cell wall components (anti-pectins JIM5, JIM7, LM19, CCRC-M38, and LM5 and anti-hemicelluloses LM25, LM15, CCRC-M1, and CCRC-M138). The localization of the examined compounds was determined using immunohistochemistry techniques, Carbotrace 680, and Calcofluor White. Our study showed the presence of various components in the cell walls of external gland cells: methylesterified and demethylesterified homogalacturonans, galactan, xylan, galactoxyloglucan, and xyloglucan. In the terminal cell, the primary cell wall contains different pectins in contrast to the secondary wall material, which is rich in cellulose and hemicelluloses. We also found that the basal cell differs from the other gland cells by the presence of galactan in the cell wall, which resembles the epidermal cells and parenchyma of traps. A particularly noteworthy part of the cell wall functions as a Casparian strip in the pedestal cell. Here, we found no labeling with Carbotrace 680, possibly due to cell wall modification or cell wall chemical composition variation. We have shown that the apoplastic space formed by the cell walls of the terminal cell is mainly composed of cellulose and hemicelluloses (galactoxyloglucan and xyloglucan). This composition of the cell walls allows the easy uptake of components from the external environment. Our research supports the external glands' function as hydropotens. [ABSTRACT FROM AUTHOR]

Pancreatic cystic neoplasms are lesions comprised of cystic components that show different biological behaviors, epidemiology, clinical manifestations, imaging features, and malignant potential and management. Benign cystic neoplasms include serous cystic neoplasms (SCAs). Other pancreatic cystic lesions have malignant potential, such as intraductal papillary mucinous neoplasms and mucinous cystic neoplasms. SCAs can be divided into microcystic (classic appearance), honeycomb, oligocystic/macrocystic, and solid patterns based on imaging appearance. They are usually solitary but may be multiple in von Hippel–Lindau disease, which may depict disseminated involvement. The variable appearances of SCAs can mimic other types of pancreatic cystic lesions, and cross-sectional imaging plays an important role in their differential diagnosis. Endoscopic ultrasonography has helped in improving diagnostic accuracy of pancreatic cystic lesions by guiding tissue sampling (biopsy) or cyst fluid analysis. Immunohistochemistry and newer techniques such as radiomics have shown improved performance for preoperatively discriminating SCAs and their mimickers. [ABSTRACT FROM AUTHOR]

Despite significant advances in prenatal medicine, spontaneous miscarriage remains one of the most common and serious pregnancy complications, affecting an increasing number of women. Since many aspects of the pathogenesis of spontaneous miscarriage remain unexplained, the aim of this study has been to assess the involvement of the NLRP3 inflammasome as a potential causative factor. The concentrations of NLRP3, IL-1β, IL-18, and cytochrome C in the serum of patients after miscarriage were measured by means of the immunoenzymatic method. In the placental tissue, the expression of NLRP3, IL-1β, IL-18, and Caspase-1 as well as that of the classical apoptosis biomarkers Fas, FasL, Bcl-2, and Ca was evaluated by means of immunohistochemistry techniques. Additionally, in whole blood, the concentrations of elements crucial for pregnancy progression, such as Ca, K, Mg, and Na, were examined by means of the ICP-OES method. Significantly higher concentrations of NLRP3 and IL-18 were demonstrated in the serum of patients with miscarriage as compared to the control group. In the placental tissue samples, a higher expression of IL-1β, IL-18, and Caspase-1 proteins was noted in women who had experienced miscarriage as compared to the control group. At the same time, a significantly lower expression of FasL and Bcl-2 proteins as well as Ca deposits was observed in women after miscarriage as compared to those with a normal pregnancy outcome. Significantly lower concentrations of Ca and K were recorded in the blood of patients with spontaneous miscarriage as compared to pregnant women. The analysis of the results x indicated a greater involvement of the inflammasome in women with spontaneous miscarriage associated with oxidative–antioxidative imbalance than in the case of miscarriage related to NET formation. Our research has provided evidence for the involvement of the inflammasome in the process of spontaneous miscarriage and identifies a new direction for diagnostics that includes NLRP3 as a preventive element in prenatal care, particularly in light of the steadily declining number of pregnancies and the increasing number of reproductive failures. [ABSTRACT FROM AUTHOR]

Background: One of the etiologies of stroke is intracerebral hemorrhage and the demand for potential therapeutic agents to alleviate symptoms and improve quality of life in patients with ICH is growing. Several phytocompounds capable of such potency with minimal adverse effects can replace drugs in market. This requires high throughput screening and in vivo experimental demonstration of the potency in ICH models that simulate mammalian pathophysiology. We aimed to study the neuroprotective activity of Asiaticoside using an adult zebrafish ICH model. Materials and Methods: Atorvastatin-induced ICH models were treated with non-toxic concentrations of Asiaticoside. Behavioural, biochemical, and cellular assays were performed to validate the developed model. Antioxidant potency was identified by testing the levels of lipid peroxidation, superoxide dismutase and xanthine oxidase in the brain homogenate. Iron chelating potency was assessed in Perl's-stained brain tissue sections. The anti-inflammatory potency was observed by measuring fold change in the expression of inflammatory gene IL-1β in brain homogenates by RT-PCR method and by immunohistochemistry techniques. The neuroprotective role of the compound was studied by fluorescent staining of the brain whole cell suspension to check apoptotic and necrotic population post-treatment. Results and Discussion: The iron accumulation in tissue sections and brain homogenates reduced 3-fold post treatment. With 600 μg/mL concentration of Asiaticoside, the malondialdehyde and xanthine levels reduced 3-fold, and scavenging activity was found to increase after 24 hr, indicating antioxidant potency. This concentration also resulted in one third reduction in the expression of pro-inflammatory cytokine IL-1β and reduced inflammatory cell infiltration showing anti-inflammatory properties. The compound was found to be neuroprotective by reducing the apoptotic cell population from 84% to 20% in comparison to untreated control. In addition, post-treatment the nitric oxide levels were restored better than the untreated control indicating modulation in cellular signalling mechanisms. Conclusion: The adult Zebrafish ICH model was developed to present the multifaceted mammalian pathophysiology of ICH in one system. The phytocompound Asiaticoside is shown to modulate neuroinflammation through a cycle of biological properties thus being neuroprotective, opening avenues for further pre-clinical testing. [ABSTRACT FROM AUTHOR]

The article in Clinical & Translational Discovery identifies a rare population of metallothionein-expressing macrophages present in all vital human organs. Using single-cell RNA sequencing data, the study characterizes these macrophages in the liver, kidney, and lung, highlighting their gene expression patterns and potential functional roles. Immunohistochemistry techniques confirm the presence of these macrophages in human liver tissue, suggesting a role in neoangiogenesis. The research sets the stage for further studies on the functional role of these macrophages in tissue homeostasis and remodeling. [Extracted from the article]

Ischemic stroke is a common and dangerous disease in clinical practice. Astrocytes (ASs) are essential for maintaining the metabolic balance of the affected regions during the disease process. 4-Hydroxybenzyl alcohol (4HBA) from Gastrodia elata Bl. has potential neuroprotective properties due to its ability to cross the blood–brain barrier. In an in vitro experiment, we replicated the oxygen–glucose deprivation/reoxygenation model, and used methyl thiazoly tertrazolium, flow cytometry, kits, and other technical means to clarify the protective effect of 4HBA on primary ASs. In in vivo experiments, the 2VO model was replicated, and immunofluorescence and immunohistochemistry techniques were used to clarify the protective effect of 4HBA on ASs and the maintenance of the blood-brain barrier. Differential metabolites and related pathways were screened and verified using metabolomics analysis and western blot. 4HBA noticeably amplified AS cell survival, reduced mitochondrial dysfunction, and mitigated oxidative stress. It demonstrated a protective effect on ASs in both environments and was instrumental in stabilizing the blood–brain barrier. Metabolomic data indicated that 4HBA regulated nucleic acid and glutathione metabolism, influencing purines, pyrimidines, and amino acids, and it activated the N-methyl-D-aspartate/p-cAMP-response element binding protein/brain-derived neurotrophic factor signaling pathway via N-methyl-D-aspartate R1/N-methyl-D-aspartate 2C receptors. Our findings suggest that 4HBA is a potent neuroprotective agent against ischemic stroke, enhancing AS cell survival and function while stabilizing the blood–brain barrier. The N-methyl-D-aspartate/p-cAMP-response element binding protein/brain-derived neurotrophic factor signaling pathway is activated by 4HBA. [ABSTRACT FROM AUTHOR]

HNRNPA2B1 is a member of the HNRNP family, which is associated with telomere function, mRNA translation, and splicing, and plays an important role in tumor development. To date, there have been no pan‐cancer studies of HNRNPA2B1, particularly within the TME. Therefore, we conducted a pan‐cancer analysis of HNRNPA2B1 using TCGA data. Based on datasets from TCGA, TARGET, Genotype‐Tissue Expression, and Human Protein Atlas, we employed a range of bioinformatics approaches to explore the potential oncogenic role of HNRNPA2B1. This included analyzing the association of HNRNPA2B1 expression with prognosis, tumor mutation burden (TMB), microsatellite instability (MSI), immune response, and immune cell infiltration of individual tumors. We further validated the bioinformatic findings using immunohistochemistry techniques. HNRNPA2B1 was found to be differentially expressed across most tumor types in TCGA's pan‐cancer database and was predictive of poorer clinical staging and survival status. HNRNPA2B1 expression was also closely linked to TMB, MSI, tumor stemness, and chemotherapy response. HNRNPA2B1 plays a significant role in the TME and is involved in the regulation of novel immunotherapies. Its expression is significantly associated with the infiltration of macrophages, dendritic cells, NK cells, and T cells. Furthermore, HNRNPA2B1 is closely associated with immune checkpoints, immune‐stimulatory genes, immune‐inhibitory genes, MHC genes, chemokines, and chemokine receptors. We performed a comprehensive evaluation of HNRNPA2B1, revealing its potential role as a prognostic indicator for patients and its immunomodulatory functions. [ABSTRACT FROM AUTHOR]

Irritable bowel syndrome (IBS), which is characterized by chronic abdominal pain, has a high global prevalence. The anterior cingulate cortex (ACC), which is a pivotal region involved in pain processing, should be further investigated regarding its role in the regulation of visceral sensitivity and mental disorders. A C57BL/6J mouse model for IBS was established using chronic acute combining stress (CACS). IBS‐like symptoms were assessed using behavioral tests, intestinal motility measurements, and abdominal withdrawal reflex scores. Fluoro‐Gold retrograde tracing and immunohistochemistry techniques were employed to investigate the projection of ACC gamma‐aminobutyric acid‐producing (GABAergic) neurons to the lateral hypothalamus area (LHA). Chemogenetic approaches enabled the selective activation or inhibition of the ACC–LHA GABAergic pathway. Enzyme‐linked immunosorbent assay (ELISA) and western blot analyses were conducted to determine the expression of histamine, 5‐hydroxytryptamine (5‐HT), and transient receptor potential vanilloid 4 (TRPV4). Our findings suggest that CACS induced IBS‐like symptoms in mice. The GABA type A receptors (GABAAR) within LHA played a regulatory role in modulating IBS‐like symptoms. The chemogenetic activation of ACC–LHA GABAergic neurons elicited anxiety‐like behaviors, intestinal dysfunction, and visceral hypersensitivity in normal mice; however, these effects were effectively reversed by the administration of the GABAAR antagonist Bicuculline. Conversely, the chemogenetic inhibition of ACC–LHA GABAergic neurons alleviated anxiety‐like behaviors, intestinal dysfunction, and visceral hypersensitivity in the mouse model for IBS. These results highlight the crucial involvement of the ACC–LHA GABAergic pathway in modulating anxiety‐like behaviors, intestinal motility alterations, and visceral hypersensitivity, suggesting a potential therapeutic strategy for alleviating IBS‐like symptoms. [ABSTRACT FROM AUTHOR]

Background: Renal cell carcinoma (RCC) is presently recognized as the most prevalent kidney tumor. However, the role and underlying mechanism of action of the conversion factor-inducible protein (TGFBI), an extracellular matrix protein, in RCC remain poorly understood. Methods: In this study, we employed Western blot, quantitative real-time polymerase chain reaction (qRT-PCR), and immunohistochemistry techniques to assess the expression of TGFBI in RCC tissues or cells. Furthermore, we analyzed the proliferation and migration of RCC cells using CCK8, cloning, scratching, and migration assays. Additionally, we examined apoptosis and cell cycle progression through flow cytometry, analysis. Lastly, we employed gene set enrichment analysis (GSEA) to investigate the biological processes associated with TGFBI, which were subsequently validated. Results: The findings indicate that TGFBI exhibits significantly elevated expression levels in both renal cell carcinoma (RCC) tissues and cells. Furthermore, the knockdown of TGFBI in SiRNA transfected cells resulted in the inhibition of RCC cell proliferation, migration, invasiveness, apoptosis, and alteration of the cell cycle. Additionally, TGFBI was found to impede the epithelial-mesenchymal transition (EMT) process in RCC cells. Bioinformatics analysis suggests that TGFBI may exert its influence on various biological processes in RCC through the tumor immune microenvironment. Moreover, our study demonstrates that TGFBI promotes RCC progression by activating the PI3K/AKT/mTOR/HIF-1α. Conclusions: Our research indicates that TGFBI exhibits high expression in RCC and facilitate RCC progression and metastasis through various molecular mechanisms. Hence, TGFBI has the potential to be a novel therapeutic target for the diagnosis and treatment of RCC in the future. [ABSTRACT FROM AUTHOR]

Egypt's meat producers produce a variety of meat brands that used soybean proteins because of high meat pricing. Allergens have been discovered in products that were not declared allergen-free. A total of 540 samples were obtained at random from various food supermarkets in Sohag City. Using haematoxylin and eosin and various histochemical dyes, light microscopy was utilized to identify the structural properties of the soybeans. The structure of all components was determined using fluorescent microscopy and acridine orange dye. Soybeans were discovered in meat samples and the cotyledon's vascular tissue and parenchymal cells rich in pectin confirmed this discovery by immunohistochemistry techniques. After immunohistochemical confirmation with an antirabbit soybean marker, the proportion of soybean was determined using histological procedures and image analysis. The percentage of soybeans in various meat products, including minced meat from two quality levels, sausages from two quality levels, raw kofta, and beef burgers from two grades, as well as chicken and luncheon meats, was 85%, 90%, 64%, 76%, 80%, 65%, 97%, 82%, and 69%, respectively. Soybean percentages in the examined samples did not fulfill Egyptian standards for soya percentages, which is approximately 10%, and this may affect consumers' health. In conclusion, all tested meat samples contained a high percentage of soya adulteration, necessitating increased supervisory to reduce meat fraud. [ABSTRACT FROM AUTHOR]

Introduction: Microsatellite instability (MSI) is a genetic marker that is useful in the detection and treatment of Lynch syndrome (Sd). Although conventional techniques such as immunohistochemistry (IHC) and polymerase chain reaction (PCR) are the standards for MSI detection, the advent of next-generation sequencing (NGS) has offered new possibilities, especially with circulating DNA. Case report: We present the case of a 26-year-old patient with Lynch Sd and a BRAF-mutated metastatic colon cancer. The discordant MSI results between the conventional methods and NGS posed challenges in making treatment decisions. Subsequent NGS analysis revealed a high MSI status, leading to participation in an immunotherapy trial, with remarkable clinical response. Conclusion: This case emphasizes the importance of comprehensive molecular profiling and strong interdisciplinary collaborations, especially in cases with ambiguous MSI results. [ABSTRACT FROM AUTHOR]

Tumor structure is heterogeneous and complex, and it is difficult to obtain complete characteristics by two‐dimensional analysis. The aim of this study was to visualize and characterize volumetric vascular information of clear cell renal cell carcinoma (ccRCC) tumors using whole tissue phenotyping and three‐dimensional light‐sheet microscopy. Here, we used the diagnosing immunolabeled paraffin‐embedded cleared organs pipeline for tissue clearing, immunolabeling, and three‐dimensional imaging. The spatial distributions of CD34, which targets blood vessels, and LYVE‐1, which targets lymphatic vessels, were examined by calculating three‐dimensional density, vessel length, vessel radius, and density curves, such as skewness, kurtosis, and variance of the expression. We then examined those associations with ccRCC outcomes and genetic alteration state. Formalin‐fixed paraffin‐embedded tumor samples from 46 ccRCC patients were included in the study. Receiver operating characteristic curve analyses revealed the associations between blood vessel and lymphatic vessel distributions and pathological factors such as a high nuclear grade, large tumor size, and the presence of venous invasion. Furthermore, three‐dimensional imaging parameters stratified ccRCC patients regarding survival outcomes. An analysis of genomic alterations based on volumetric vascular information parameters revealed that PI3K‐mTOR pathway mutations related to the blood vessel radius were significantly different. Collectively, we have shown that the spatial elucidation of volumetric vasculature information could be prognostic and may serve as a new biomarker for genomic alterations. High‐end tissue clearing techniques and volumetric immunohistochemistry enable three‐dimensional analysis of tumors, leading to a better understanding of the microvascular structure in the tumor space. [ABSTRACT FROM AUTHOR]

Purpose: Lung cancer is the leading cause of cancer-related deaths worldwide. However, with the optimization of screening strategies and advances in treatment, mortality has been decreasing in recent years. In this study, we describe non-small cell lung cancer patients diagnosed between 2021 and 2022 at a high-complexity hospital in Latin America, as well as the immunohistochemistry techniques used to screen for ROS1 rearrangements, in the context of the recent approval of crizotinib for the treatment of ROS1 rearrangements in non-small cell lung cancer in Colombia. Methods: A descriptive cross-sectional study was conducted. Sociodemographic, clinical, and molecular pathology information from non-small cell lung cancer individuals who underwent immunohistochemistry to detect ROS1 rearrangements between 2021 and 2022 at Fundación Valle del Lili (Cali, Colombia) was recorded. The clinical outcomes of confirmed ROS1 rearrangements in non-small cell lung cancer patients were reported. Results: One hundred and thirty-six patients with non-small cell lung cancer were included. The median age at diagnosis was 69.8 years (interquartile range 61.9– 77.7). At diagnosis, 69.8% (n = 95) were at stage IV. ROS1 immunohistochemistry was performed using the monoclonal D4D6 antibody clone in 54.4% (n = 74) of the cases, while 45.6% (n = 62) were done with the monoclonal SP384 antibody clone. Two patients were confirmed to have ROS1 rearrangements in non-small cell lung cancer using next-generation sequencing and received crizotinib. On follow-up at months 5.3 and 7.0, one patient had a partial response, and the other had oligo-progression, respectively. Conclusion: Screening for ROS1 rearrangements in non-small cell lung cancer is imperative, as multiple prospective studies have shown improved clinical outcomes with tyrosine kinase inhibitors. Given the recent approval of crizotinib in Colombia, public health policies must be oriented toward early detection of driver mutations and prompt treatment. Additionally, future approvals of newly tested tyrosine kinase inhibitors should be anticipated. [ABSTRACT FROM AUTHOR]

Chronic kidney disease (CKD) poses a significant global health dilemma, emerging from complex causes. Although our prior research has indicated that a deficiency in Reticulon-3 (RTN3) accelerates renal disease progression, a thorough examination of RTN3 on kidney function and pathology remains underexplored. To address this critical need, we generated Rtn3-null mice to study the consequences of RTN3 protein deficiency on CKD. Single-cell transcriptomic analyses were performed on 47,885 cells from the renal cortex of both healthy and Rtn3-null mice, enabling us to compare spatial architectures and expression profiles across 14 distinct cell types. Our analysis revealed that RTN3 deficiency leads to significant alterations in the spatial organization and gene expression profiles of renal cells, reflecting CKD pathology. Specifically, RTN3 deficiency was associated with Lars2 overexpression, which in turn caused mitochondrial dysfunction and increased reactive oxygen species levels. This shift induced a transition in renal epithelial cells from a functional state to a fibrogenic state, thus promoting renal fibrosis. Additionally, RTN3 deficiency was found to drive the endothelial-to-mesenchymal transition process and disrupt cell-cell communication, further exacerbating renal fibrosis. Immunohistochemistry and Western-Blot techniques were used to validate these observations, reinforcing the critical role of RTN3 in CKD pathogenesis. The deficiency of RTN3 protein in CKD leads to profound changes in cellular architecture and molecular profiles. Our work seeks to elevate the understanding of RTN3's role in CKD's narrative and position it as a promising therapeutic contender. [ABSTRACT FROM AUTHOR]

Species in the genus Utricularia are carnivorous plants that prey on invertebrates using traps of leaf origin. The traps are equipped with numerous different glandular trichomes. Trichomes (quadrifids) produce digestive enzymes and absorb the products of prey digestion. The main aim of this study was to determine whether arabinogalactan proteins (AGPs) occur in the cell wall ingrowths in the quadrifid cells. Antibodies (JIM8, JIM13, JIM14, MAC207, and JIM4) that act against various groups of AGPs were used. AGP localization was determined using immunohistochemistry techniques and immunogold labeling. AGPs localized with the JIM13, JIM8, and JIM14 epitopes occurred in wall ingrowths of the pedestal cell, which may be related to the fact that AGPs regulate the formation of wall ingrowths but also, due to the patterning of the cell wall structure, affect symplastic transport. The presence of AGPs in the cell wall of terminal cells may be related to the presence of wall ingrowths, but processes also involve vesicle trafficking and membrane recycling, in which these proteins participate. [ABSTRACT FROM AUTHOR]

Renal cell carcinoma (RCC), accounting for 90% of all kidney cancer, is categorized into clear cell RCC (ccRCC) and non-clear cell RCC (non-ccRCC) for treatment based on the current NCCN Guidelines. Thus, the classification will be associated with therapeutic implications. This study aims to identify novel biomarkers to differentiate ccRCC from non-ccRCC using bioinformatics and machine learning. The gene expression profiles of ccRCC and non-ccRCC subtypes (including papillary RCC (pRCC) and chromophobe RCC (chRCC)), were obtained from TCGA. Differential expression genes (DEGs) were identified, and specific DEGs for ccRCC and non-ccRCC were explored using a Venn diagram. Gene Ontology and pathway enrichment analysis were performed using DAVID. The top ten expressed genes in ccRCC were then selected for machine learning analysis. Feature selection was operated to identify a minimum highly effective gene set for constructing a predictive model. The expression of best-performing gene set was validated on tissue samples from RCC patients using immunohistochemistry techniques. Subsequently, machine learning models for diagnosing RCC were developed using H-scores. There were 910, 415, and 835 genes significantly specific for DEGs in ccRCC, pRCC, and chRCC, respectively. Specific DEGs in ccRCC enriched in PD-1 signaling, immune system, and cytokine signaling in the immune system, whereas TCA cycle and respiratory, signaling by insulin receptor, and metabolism were enriched in chRCC. Feature selection based on Decision Tree Classifier revealed that the model with two genes, including NDUFA4L2 and DAT, had an accuracy of 98.89%. Supervised classification models based on H-score of NDUFA4L2, and DAT revealed that Decision Tree models showed the best performance with 82% accuracy and 0.9 AUC. NDUFA4L2 expression was associated with lymphovascular invasion, pathologic stage and pT stage in ccRCC. Using integrated bioinformatics and machine learning analysis, NDUFA4L2 and DAT were identified as novel biomarkers to differential diagnosis ccRCC from non-ccRCC. [ABSTRACT FROM AUTHOR]

Background: Glioma is the main subtype of primary central nervous system (CNS) tumor with high malignancy and poor prognosis under current therapeutic approaches. Glycolysis and suppressive tumor microenvironment (TME) are key markers of glioma with great importance for aggressive features of glioma and inferior clinical outcomes. Hexokinase 3 (HK3) is an important rate-limiting enzyme in glycolysis, but its function in glioma remains unknown. Methods: This study comprehensively assessed the expression distribution and immunological effect of HK3 via pan-cancer analysis based on datasets from Genotype Tissue Expression (GTEx), Cancer Cell Line Encyclopedia (CCLE), and The Cancer Genome Atlas (TCGA). Furthermore, it explored the malignant phenotype and genomic landscape between low-HK3 and high-HK3 expression groups in gliomas from Chinese Glioma Genome Atlas (CGGA) and TCGA. Moreover, data from the TIMER website predicted the relationship between macrophage infiltration and HK3 expression. Also, single-cell sequencing data were used to validate the relationship. Results: For pan-cancer patients, HK3 was expressed in various cancers. The results showed that HK3 was highly expressed in gliomas and positively correlated with tumor-infiltrating immune cells (TIICs), immune checkpoints, immunomodulators, and chemokines. Meanwhile, HK3 expression was highest in normal immune cells and tissues. In gliomas, the expression of HK3 was found to be closely correlated with the malignant clinical characteristics and the infiltration of macrophages. Also, HK3 was proven to be positively associated with macrophage through single-cell sequencing data and immunohistochemistry techniques. Finally, it is predicted that samples with high HK3 expression are often malignant entities and also significant genomic aberrations of driver oncogenes. Conclusions: This is the first comprehensive research to figure out the relationship between HK3 and TME characteristics in gliomas. HK3 is positively associated with macrophage infiltration and can induce the immunosuppressive TME and malignant phenotype of gliomas. [ABSTRACT FROM AUTHOR]

The genus Utricularia (bladderworts) species are carnivorous plants that prey on invertebrates using traps with a high-speed suction mechanism. The outer trap surface is lined by dome-shaped glands responsible for secreting water in active traps. In terminal cells of these glands, the outer wall is differentiated into several layers, and even cell wall ingrowths are covered by new cell wall layers. Due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls (microdomains). The main aim of this study was to check if different cell wall layers have a different composition. Antibodies against arabinogalactan proteins (AGPs) were used, including JIM8, JIM13, JIM14, MAC207, and JIM4. The localization of the examined compounds was determined using immunohistochemistry techniques and immunogold labeling. Differences in composition were found between the primary cell wall and the cell secondary wall in terminal gland cells. The outermost layer of the cell wall of the terminal cell, which was cuticularized, was devoid of AGPs (JIM8, JIM14). In contrast, the secondary cell wall in terminal cells was rich in AGPs. AGPs localized with the JIM13, JIM8, and JIM14 epitopes occurred in wall ingrowths of pedestal cells. Our research supports the hypothesis of water secretion by the external glands. [ABSTRACT FROM AUTHOR]

Endometrial cancer (EC) accounts for 90% of uterine cancer cases. It is considered not only one of the most common gynecological malignancies but also one of the most frequent cancers among women overall. Nowadays, the differentiation of EC subtypes is based on immunohistochemistry and molecular techniques. It is considered that patients' prognosis and the implementation of the appropriate treatment depend on the cancer subtype. Patients with pathogenic variants in POLE have the most favorable outcome, while those with abnormal p53 protein have the poorest. Therefore, in patients with POLE mutation, the de-escalation of postoperative treatment may be considered, and patients with abnormal p53 protein should be subjected to intensive adjuvant therapy. Patients with a DNA mismatch repair (dMMR) deficiency are classified in the intermediate prognosis group as EC patients without a specific molecular profile. Immunotherapy has been recognized as an effective treatment method in patients with advanced or recurrent EC with a mismatch deficiency. Thus, different adjuvant therapy approaches, including targeted therapy and immunotherapy, are being proposed depending on the EC subtype, and international guidelines, such as those published by ESMO and ESGO/ESTRO/ESP, include recommendations for performing the molecular classification of all EC cases. The decision about adjuvant therapy selection has to be based not only on clinical data and histological type and stage of cancer, but, following international recommendations, has to include EC molecular subtyping. This review describes how molecular classification could support more optimal therapeutic management in endometrial cancer patients. [ABSTRACT FROM AUTHOR]

Bactrian camels inhabiting desert and semi-desert regions of China are valuable animal models for studying adaptation to desert environments and heat stress. In this study, 16S rRNA technology was employed to investigate the distribution characteristics and differences of mucosal microorganisms in the anterior gland area, posterior gland area, third gland area, cardia gland area, gastric fundic gland area and pyloric gland area of 5-peak adult healthy Bactrian camels. We aimed to explore the possible reasons for the observed microbial distribution from the aspects of histological structure and mucosal immunity. Bacteroides and Fibrobacteria accounted for 59.54% and 3.22% in the gland area, respectively, and 52.37% and 1.49% in the wrinkled stomach gland area, respectively. The gland area showed higher abundance of Bacteroides and Fibrobacteria than the wrinkled stomach gland area. Additionally, the anterior gland area, posterior gland area, third gland area, and cardia gland area of Bactrian camels mainly secreted acidic mucus, while the gastric fundic gland area mainly secreted neutral mucus and the pyloric region mainly secreted a mixture of acidic and neutral mucus. The results of immunohistochemistry techniques demonstrated that the number of IgA+ cells in the anterior glandular area, posterior glandular area, third glandular area, and cardia gland area was significantly higher than that in the fundic and pyloric gland area (p < 0.05), and the difference in IgA+ between the fundic and pyloric gland area was not significant (p > 0.05). The study revealed a large number of bacteria that can digest and degrade cellulose on the mucosa of the gastric gland area of Bactrian camels. The distribution of IgA+ cells, the structure of the mucosal tissue in the glandular region, and the composition of the mucus secreted on its surface may have a crucial influence on microbial fixation and differential distribution. [ABSTRACT FROM AUTHOR]

Ulcerative colitis (UC), as a chronic inflammatory disease, presents a global public health threat. However, the mechanism of Poria cocos (PC) in treating UC remains unclear. Here, LC-MS/MS was carried out to identify the components of PC. The protective effect of PC against UC was evaluated by disease activity index (DAI), colon length and histological analysis in dextran sulfate sodium (DSS)-induced UC mice. ELISA, qPCR, and Western blot tests were conducted to assess the inflammatory state. Western blotting and immunohistochemistry techniques were employed to evaluate the expression of tight junction proteins. The sequencing of 16S rRNA was utilized for the analysis of gut microbiota regulation. The results showed that a total of fifty-two nutrients and active components were identified in PC. After treatment, PC significantly alleviated UC-associated symptoms including body weight loss, shortened colon, an increase in DAI score, histopathologic lesions. PC also reduced the levels of inflammatory cytokines TNF-α, IL-6, and IL-1β, as evidenced by the suppressed NF-κB pathway, restored the tight junction proteins ZO-1 and Claudin-1 in the colon, and promoted the diversity and abundance of beneficial gut microbiota. Collectively, these findings suggest that PC ameliorates colitis symptoms through the reduction in NF-κB signaling activation to mitigate inflammatory damage, thus repairing the intestinal barrier, and regulating the gut microbiota. [ABSTRACT FROM AUTHOR]

Background: Dendritic cells participate in the pathophysiology of lupus erythematosus (LE), which are studied in systemic and cutaneous forms; however, little is known about their oral manifestations. Methods: The expressions of dendritic cell markers (including CD1a, CD21, CD123, and langerin) were investigated by immunohistochemistry technique. Sixty intraoral and lower lip LE lesions, and additional 10 control samples were collected from 2003 to 2019. They were topographically analyzed in the epithelium (EP), lamina propria (LP), epithelial junction (JUN), and deep perivascular (PV) areas. Results: The expression of CD1a was decreased in the EP (p = 0.003) and increased in the deep PV area (p = 0.002). Langerin immunostaining showed no significant decrease in EP (p = 0.944); however, it increased in LP (p = 0.012) and JUN (p = 0.006). CD21 was expressed in only two specimens (EP, p = 0.012; LP, p < 0.001; deep PV area, p = 0.018). CD123 expression increased in all topographies (EP, p < 0.005; LP, p < 0.001, JUN, p < 0.001; deep PV, p < 0.001). The comparison between vermilion and intraoral mucosa LE lesions suggested that sun‐exposed sites showed higher expression of CD123 (EP, p = 0.024; LP, p = 0.047; JUN, p = 0.001). Conclusions: CD1a, langerin, and CD123 expressions were detected coincidently surrounding the inflammatory infiltrate in oral LE, suggesting that these cells may play an important role in immune response. Interestingly, plasmacytoid dendritic cells showed increased CD123 expression in sun‐exposed site lesions, which point out a possible function in their pathogenesis. Further studies are needed to confirm this hypothesis. [ABSTRACT FROM AUTHOR]

Background. Urinary bladder cancer is a prevalent global health concern. IL-32 is a cytokine that promotes inflammation and has been shown in multiple studies to suppress the development of cancer cells and trigger cell death in different types of cancer cells. Materials and methods. Expression of 32 interleukins was determined using an immunohistochemistry technique, which uses analysis of the exact concentrations and locations of specific compounds in tissues. This is done by using antibodies, antibodies that react with the target protein (in this case IL-32), then identifying the sites where the active protein is located. The study included 114 diagnosed bladder cancer samples and 10 healthy controls. Results. By analyzing the expression of IL-32 using IHC technology on both bladder tissue from cancer patients and healthy bladder tissue that was considered a control, the results of evaluating IL-32 levels in bladder tissue from cancer patients showed a high level. Conclusion. High levels of IL-32 indicate disease progression or a poor outcome, while low levels can indicate an improvement or a good outcome. This is consistent with the fundamental aim of this study: the possibility of using IL-32 as a diagnostic or prognostic marker for disease progression and outcome prediction in cancer patients. [ABSTRACT FROM AUTHOR]

Introduction: Coronary artery disease (CAD) is a major and multifaceted health problem but also the first cause of death in modern Western societies. Furthermore, myocardial infarction (MI) constitutes a challenge for analysis in the field of molecular mechanisms, early diagnosis and therapeutic approaches, as its incidence increases every year worldwide. Concerning the histopathological diagnosis in the corresponding cases, a variety of immunohistochemistry (IHC) markers and methods are available to support conventional histology diagnosis. Immunohistochemistry techniques are effective for use in forensic pathology, expanding the limits of differential diagnoses in borderline cases, as they can be applied to tissue samples fixed in formalin and embedded in paraffin. Objective: The purpose of the current review was to explore the role of connexin 43 (gene locus: 6q22.31) as a reliable biomarker of myocardial disease/infarction and its impact on MI pathology. Material and method: A systematic review of the literature was carried out based on the international database PubMed. The majority of medical data referred to articles published after the year 2020, whereas specific references of great importance and value were also included. The following keywords were used: coronary, artery, myocardial, infarction, connexin and immunohistochemistry. Results: A pool of 38 significant articles focused on the mechanisms and novel experimental biomarkers was selected for the present study at the basis of combining molecular knowledge with new clinical features in CAD, and MI histodiagnosis. Conclusions: The role of connexin 43 -- as a significant gap junction intermediate protein -- in MI pathology, clinical symptoms and prognosis is critical because its dysfunction is involved in myocardial conduction and the onset of ventricular arrhythmias due to a crucial interruption of the intra-cardiomyocyte's conjunction. [ABSTRACT FROM AUTHOR]

A key component of amphibian antipredator strategies is the chemical defenses that make them toxic and/or distasteful, like the cardiotoxic bufadienolides synthesized by the true toads and stored in granular skin glands. The morphology of adult toad glands is well-described, but the ontogenetic timing and distribution of glands during larval stages are poorly understood. A comparative understanding of granular gland development is needed to elucidate the mechanisms underlying the diversification of toad chemical defenses and their ecological roles across lineages and ontogeny. Herein, we analyzed gland morphology before and after metamorphosis of Rhinella horribilis, Incilius melanochlorus, and I. luetkenii. We hypothesized that granular gland development would begin earlier and progress faster in relatively more toxic Rhinella species. Our results showed that the timeline of skin development, the relative dimensions and quantity of structures, and the appearance of protein and mucin content in granular and mucous glands did not vary among Rhinella and Incilius. Furthermore, epidermal mucus and/or giant cells in larval toads may act as sources of chemical defenses prior to gland development. Our findings suggest that gland development is well-conserved among these genera; it is possible that reported differences in toxin profiles are not due to divergent morphogenesis during larval or metamorphic stages, but rather to differences in molecular function or structural differentiation in juveniles or adults. Therefore, complementary studies using integrative techniques such as immunohistochemistry and comparative transcriptomics are needed to uncover the mechanisms responsible for the diversity of chemical defenses found in bufonid toads across development. [ABSTRACT FROM AUTHOR]

Blood flow restriction applied during low-load resistance training (LL-BFR) induces a similar increase in the cross-sectional area of muscle fibers (fCSA) compared to traditional high-load resistance training (HL-RT). However, it is unclear whether LLBFR leads to differential changes in myofibrillar spacing in muscle fibers and/or extracellular area compared to HL-RT. Therefore, this study aimed to investigate whether the hypertrophy of type I and II fibers induced by LL-BFR or HL-RT is accompanied by differential changes in myofibrillar and non-myofibrillar areas. In addition, we examined if extracellular spacing was differentially affected between these two training protocols. Twenty recreationally active participants were assigned to LL-BFR or HL-RT groups and underwent a 6-week training program. Muscle biopsies were taken before and after the training period. The fCSA of type I and II fibers, the area occupied by myofibrillar and non-myofibrillar components, and extracellular spacing were analyzed using immunohistochemistry techniques. Despite the significant increase in type II and mean (type I + II) fCSA (p < 0.05), there were no significant changes in the proportionality of the myofibrillar and nonmyofibrillar areas [~86% and ~14%, respectively (p > 0.05)], indicating that initial adaptations to LL-BFR are primarily characterized by conventional hypertrophy rather than disproportionate non-myofibrillar expansion. Additionally, extracellular spacing was not significantly altered between protocols. In summary, our study reveals that LL-BFR, like HL-RT, induces skeletal muscle hypertrophy with proportional changes in the areas occupied by myofibrillar, non-myofibrillar, and extracellular components. [ABSTRACT FROM AUTHOR]

Psoriasis is a recurrent and protracted disease that severely impacts the patient's physical and mental health. Thus, there is an urgent need to explore its pathogenesis to identify therapeutic targets. The expression level of protein tyrosine phosphatase nonreceptor type 2 (PTPN2) was analyzed by immunohistochemistry techniques in psoriatic tissues and imiquimod‐induced psoriatic mouse models. PTPN2 and signal transducer and activator of transcription 3 (STAT3) were overexpressed or silenced in human keratinocytes or an interleukin (IL)‐6‐induced psoriasis HaCaT cell model using overexpression plasmid transfection or small interfering RNA technology in vitro, and the effects of PTPN2 on STAT3, HaCaT cell function, and autophagy levels were investigated using reverse transcription‐quantitative polymerase chain reaction, Western blot, Cell Counting Kit 8, 5‐ethynyl‐20‐deoxyuridine, flow cytometry, and transmission electron microscopy. PTPN2 expression was found to be significantly downregulated in psoriatic tissues. Then, the in vitro antipsoriatic properties of PTPN2 were investigated in an IL‐6‐induced psoriasis‐like cell model, and the results demonstrated that inhibition of keratinocyte proliferation by PTPN2 may be associated with elevated STAT3 dephosphorylation and autophagy levels. These findings provide novel insights into the mechanisms of autophagy in psoriatic keratinocytes and may be essential for developing new therapeutic strategies to improve inflammatory homeostasis in psoriatic patients. Significance statement: Psoriasis is a recurrent and chronic systemic inflammatory skin disease, which seriously affects the physical and mental health of patients. In this manuscript, we report for the first time that protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is significantly underexpressed in psoriasis. PTPN2 mediates signal transducer and activator of transcription 3 dephosphorylation, promoting autophagy levels and inhibiting keratinocyte proliferation. To the best of our knowledge, psoriasis cannot be adequately controlled even though the clinical efficacy of the present treatment for the etiology of the condition has greatly improved. The search explores the mechanism of pathogenesis of PTPN2 in psoriasis, providing new insights into the mechanism of autophagy in psoriatic keratinocytes, and may have important implications for the development of new therapeutic strategies to improve inflammatory homeostasis in patients with psoriasis. [ABSTRACT FROM AUTHOR]

Overactive bladder (OAB) is an age-related disorder characterised by unstable bladder contractions resulting in disruptive lower urinary tract symptoms (LUTS), thus creating a profound impact on an individual's quality of life. The development of LUTS may be linked to the overexpression of oxytocin receptors (OXTRs) within the bladder detrusor muscle, resulting in increased baseline myogenic tone. Thus, it is hypothesised that targeting OXTRs within the bladder using oxytocin antagonists may attenuate myogenic tone within the bladder, thereby providing a new therapeutic avenue for treating OAB. Organ bath contractility and immunohistochemistry techniques were conducted on bladder tissue sourced from young rats (7–8 weeks and 10–12 weeks) and older rats (4–5 months and 7–9 months). Organ bath studies revealed that oxytocin (OT) significantly increased bladder contractions, which were significantly attenuated by [β-Mercapto-β,β-cyclopentamethylenepropionyl1, O-Me-Tyr2, Orn8]-Oxytocin) (1 µM) (**** p < 0.0001) and atosiban (10 µM) in both young and older rats (** p < 0.01); in contrast, cligosiban (1 µM and 10 µM) did not inhibit OT-induced contractions in both young and older rats (p ≥ 0.05). Interestingly, cligosiban (1 µM and 10 µM) significantly reduced the frequency of spontaneous contractions within the bladder of both young (*** p < 0.001) and older rats (**** p < 0.0001), while atosiban (10 µM) only demonstrated this effect in older rats (** p < 0.01). Furthermore, immunohistochemistry (IHC) analysis revealed significant colocalization of nuclear-specific oxytocin receptors (OXTRs) in the contractile (smooth muscle) cells within young (** p < 0.01) and older rats (* p < 0.05), indicating OT may be a key modulator of bladder contractility. [ABSTRACT FROM AUTHOR]

Key Clinical Message: The clinical presentations and pathological features of low‐grade myxofibrosarcoma can be misleading, frequently resulting in diagnostic errors. An accurate diagnosis requires the application of immunohistochemistry techniques and the discerning diagnostic acumen of experienced pathologists. A 62‐year‐old male patient visited our outpatient clinic with multiple painful and rapidly enlarging subcutaneous nodules on his right forearm. Initially, the condition was misdiagnosed as multiple lipomas. The final pathology revealed characteristics consistent with low‐grade myxofibrosarcoma. [ABSTRACT FROM AUTHOR]

Wound age estimation is a significant issue in forensic pathology. Although various methods have been evaluated, no gold standard system or model has been proposed, and accurate injury time estimation is still challenging. The distinction between vital skin wounds—i.e., ante-mortem lesions—and skin alterations that occur after death is a crucial goal in forensic pathology. Once the vitality of the wound has been confirmed, the assessment of the post-trauma interval (PTI) is also fundamental in establishing the causal relationship between the traumatic event and death. The most frequently used techniques in research studies are biochemistry, molecular biology, and immunohistochemistry (IHC). Biochemical methods take advantage of the chemical and physical techniques. A systematic literature search of studies started on 18 February 2023. The search was conducted in the main databases for biomedical literature, i.e., PubMed and Scopus, for papers published between 1973 and 2022, focusing on different techniques of immunohistochemistry and immunofluorescence (IF) for estimating the PTI of skin wounds. The present study involves a comprehensive and structured analysis of the existing literature to provide a detailed and comprehensive overview of the different IHC techniques used to date skin lesions, synthesize the available evidence, critically evaluate the methodologies, and eventually draw meaningful conclusions about the reliability and effectiveness of the different markers that have been discovered and used in wound age estimation. [ABSTRACT FROM AUTHOR]

Simple Summary: Cadmium (Cd) is a major aquatic pollutant that can cause toxic effects on aquatic animals, including the economically important clam Meretrix meretrix. A transcriptomic analysis was performed on M. meretrix fertilized eggs, D-shaped larvae, pediveligers, and postlarvae following exposure to Cd2+-containing seawater. A number of genes related to the oxidative stress response were found to be differentially expressed in the fertilized egg and the different larval forms when exposed to Cd2+. These included the CCO, Ndh, HPX, A2M, STF, and pro-C3 genes, and changes in their expression levels in response to Cd2+ exposure were confirmed by qRT-PCR assays. In addition, qRT-PCR also revealed marked changes in the expression of two other oxidative stress-related genes, Nrf2 and MT, in the fertilized egg and different larval forms of M. meretrix in response to Cd2+ exposure. The results suggested that changes in oxidative stress-related genes might be crucial for M. meretrix to cope with the adverse effects of Cd during its development. Cadmium (Cd) is one of the major pollutants in the aquatic environment, and it can easily accumulate in aquatic animals and result in toxic effects by changing the metabolism of the body, causing a serious impact on the immune system, reproductive system, and the development of offspring. The clam Meretrix meretrix is one of the commercially important species that is cultivated in large-scale aquaculture in China. To elucidate the underlying molecular mechanisms of Cd2+ in the developmental processes, fertilized eggs and larvae of M. meretrix at different developmental stages were exposed to Cd2+ (27.2 mg L−1 in natural seawater) or just natural seawater without Cd2+ (control), and high-throughput transcriptome sequencing and immunohistochemistry techniques were used to analyze the toxic effects of Cd on larvae at different early developmental stages. The results revealed 31,914 genes were differentially expressed in the different stages of M. meretrix development upon treatment with Cd2+. Ten of these genes were differentially expressed in all stages of development examined, but they comprised only six unigenes (CCO, Ndh, HPX, A2M, STF, and pro-C3), all of which were related to the oxidative stress response. Under Cd exposure, the expression levels of CCO and Ndh were significantly upregulated in D-shaped and pediveliger larvae, while pro-C3 expression was significantly upregulated in the fertilized egg, D-shaped larva, and pediveliger. Moreover, HPX, A2M, and STF expression levels in the fertilized egg and pediveliger larvae were also significantly upregulated. In contrast, CCO, Ndh, HPX, A2M, STF, and pro-C3 expression levels in the postlarva were all downregulated under Cd exposure. Besides the genes with changes in expression identified by the transcriptome, the expression of two other oxidative stress-related genes (MT and Nfr2) was also found to change significantly in the different developmental stages of M. meretrix upon Cd exposure, confirming their roles in combating oxidative stress. Overall, the findings of this study indicated that Cd would interfere with cellular respiration, ion transport, and immune response through inducing oxidative stress, and changes in the expression of oxidative stress-related genes might be an important step for M. meretrix to deal with the adverse effects of Cd at different stages of its development. [ABSTRACT FROM AUTHOR]

In a scenario where there is an increased incidence of oral squamous cell carcinoma (OSCC) in younger age groups, the diagnosis of pseudo-malignant lesions that mimic the histopathology of a moderate or even high-grade carcinoma becomes imperative for oral pathologists. Though paediatric malignancies such as melanomas and thyroid carcinomas and even OSCCs have been reported in young children, they are rare in the pre-pubertal age group. Melanocytic naevi such as Spitz naevi (SNs) or atypical SNs is, however, more common in this age group and could create some difficulty in diagnosis due to its histological variations that could mimic a malignancy. Hence, the need for a cautious correlation between clinical and histopathological features becomes manifold. Adjunct tools that use diagnostic and molecular techniques such as immunohistochemistry (IHC) and comparative genomic hybridisation (CGH) help in diagnosis and in differentiating certain types of SNs from Spitzoid melanomas or melanomas. A case that histopathologically resembled a moderately differentiated squamous cell carcinoma without any evidence of melanocytic content proved to be a melanocytic naevus after clinical correlation of both age and immunohistochemical analysis. This case report with review brings to light the importance of being aware of such pseudo-malignant lesions in our daily practice. [ABSTRACT FROM AUTHOR]

Introduction: Cytological examination of serous fluids is one of the commonly performed investigation. It is crucial to distinguish between benign and malignant effusions, as it is important for determining the prognosis and treatment of the patient. Diagnostic problem arises in everyday practice to differentiate reactive atypical mesothelial cells from malignant cells by routine conventional smear method. Cytocentrifuge will provide better quality smears for interpretation and cell block technique provides better architectural pattern, morphological features and an additional yield of malignant cells, and thereby, increasing the sensitivity of the cytodiagnosis when compared to conventional smears. Aim: To compare the cytomorphological features of aspirates from body cavities using conventional smear, cytospin, cell block technique and perform IHC stains on cell block material in the diagnosis of malignant serous effusion. Materials and Methods: A total of 262 serous fluid samples over a period of two years, were subjected to simultaneous processing by conventional smear, cytocentrifuge and cell block technique. Results were compared for general cytological, cellular features and diagnostic utility for malignancy. Results: Samples comprised of 172 pleural, 82 peritoneal and 8 pericardial effusions. Conventional smear and cell block provided significantly better staining quality and morphological features. Cell block and cytospin cytology provided significantly high cellularity. Minimal overlapping of cells were significantly seen in cytocentrifuge smears. Additional yield for malignancy was 4% more by cell block method. Conclusion: The cell block technique not only increased the positive results for malignancy, but also helped to demonstrate better architectural patterns, which could be of great help in making correct diagnosis of the primary site. The cell block technique was also useful for special stains and immunohistochemistry. Cell block with ancillary technique like immunohistochemistry enhanced diagnostic accuracy. [ABSTRACT FROM AUTHOR]

The article discusses the predictors of dupilumab prescription in patients with chronic rhinosinusitis with nasal polyps. The study, conducted by researchers from Vanderbilt University Medical Center and the University of Alabama-Birmingham, aimed to identify factors influencing the prescription of dupilumab. The study found that patients with higher postoperative SNOT-22 scores were more likely to be prescribed dupilumab, indicating a correlation between symptom severity and medication prescription. The researchers acknowledged limitations in their study design, such as the cross-sectional analysis and short follow-up timeframe, and suggested that further research with larger sample sizes and longer follow-up periods could provide more comprehensive insights into dupilumab prescription trends. [Extracted from the article]

Histone deacetylation is an important mechanism involved in human breast cancer tumorigenesis and recent veterinary oncology studies also demonstrate a similar relationship in some canine neoplasms. The use of HDAC inhibitors in vitro and in vivo has demonstrated antitumor action on several strains of human and animal cancers. The present study aims to correlate the expression of H3K9Ac, H4K12Ac, HDAC1, HDAC2 and HDAC6 in simple mammary carcinomas in dogs with clinicopathological parameters and overall survival time. To this end, 61 samples of simple breast carcinomas were analyzed by the immunohistochemistry technique with subsequent validation of the antibodies by the Western Blot technique. The expressions obtained via a semi-quantitative way were categorized by assigning scores and classified into high or low expressions according to the given score, except for HDAC6, when the marking percentage was considered and subdivided into high and low expressions using the median value. For statistical analysis, the chi-square test or Fisher exact test were used as univariate analysis and correspondence analysis as a multivariate test, in addition to the Kaplan-Meier survival analysis. In the studied samples, the highest frequencies were determined for the high expression proteins H4K12Ac (88.5%), HDAC2 (65.6%) and HDAC6 (56.7%) and the low expression proteins H3K9Ac (73.8%) and HDAC1 (54.1%). An association between the low expression of HDAC1 and the presence of lymph node metastasis (p = 0.035) was indicated by univariate analysis while the high expression of HDAC1 was associated with favorable prognostic factors, such as the absence of lymph node metastasis and low mitotic index by multivariate analysis. Also, by multivariate analysis, the low expression of HDAC6 was correlated with the low expression of Ki67, smaller tumors, and better prognosis factors as well. Protein expression was not correlated with patients' overall survival time (p > 0.05). The high expressions of HDAC2 and HDAC6 in mammary carcinomas in female dogs may be useful information for research involving therapeutic targets with iHDACs since their inhibition favors hyperacetylation and transcription of tumor suppressor genes. [ABSTRACT FROM AUTHOR]

Background: Autonomic remodeling of the atria plays a pivotal role in the development of atrial fibrillation (AF) and exerts a substantial influence on the progression of this condition. Hyperlipidemia is a predisposing factor for AF, but its effect on atrial nerve remodeling is unclear. The primary goal of this study was to explore the possible mechanisms through which the consumption of a high-fat diet (HFD) induces remodeling of atrial nerves, and to identify novel targets for clinical intervention. Methods: Cell models were created in vitro by subjecting cells to palmitic acid (PA), while rat models were established by feeding them a high-fat diet. To investigate the interplay between cardiomyocytes and nerve cells in a co-culture system, we utilized Transwell cell culture plates featuring a pore size of 0.4 μm. The CCK-8 assay was employed to determine cell viability, fluorescent probe DCFH-DA and flow cytometry were utilized for measuring ROS levels, JC-1 was used to assess the mitochondrial membrane potential, the Griess method was employed to measure the nitric oxide (NO) level in the supernatant, a fluorescence-based method was used to measure ATP levels, and MitoTracker was utilized for assessing mitochondrial morphology. The expression of pertinent proteins was evaluated using western blotting (WB) and immunohistochemistry techniques. SNAP was used to treat nerve cells in order to replicate a high-NO atmosphere, and the level of nitroso was assessed using the iodoTMT reagent labeling method. Results: The study found that cardiomyocytes' mitochondrial morphology and function were impaired under high-fat stimulation, affecting nitric oxide (NO) production through the CRIF1/SIRT1/eNOS axis. In a coculture model, overexpression of eNOS in cardiomyocytes increased NO expression. Moreover, the increased Keap1 nitrosylation within neuronal cells facilitated the entry of Nrf2 into the nucleus, resulting in an augmentation of P21 transcription and a suppression of proliferation. Atrial neural remodeling occurred in the HFD rat model and was ameliorated by increasing myocardial tissue eNOS protein expression with trimetazidine (TMZ). Conclusions: Neural remodeling is triggered by high-fat stimulation, which decreases the production of NO through the CRIF1/eNOS/P21 axis. Additionally, TMZ prevents neural remodeling and reduces the occurrence of AF by enhancing eNOS expression. [ABSTRACT FROM AUTHOR]

The aim of this study is to determine the expression of the PD_L1protein in human bladder cancer, and significant correlations between these parameters and clinico pathologic variables like (grade and stage), and also by using this marker can classify bladder cancer in to basal and luminal type. the bladder cancer is one of the most common cancers worldwide. the incidence and mortality of this disease increased during last decades alarmingly in Iraq. The current study is designed to detect the role of pD_L1 expression in bladder carcinoma as a possible marker for detecting the biological behavior of malignancy and its correlation with grade and muscle invasiveness for both diagnostic and prognostic purposes. The study focuses on a technique of immunohistochemistry for detection pD_L1 expression in bladder cancer. The samples are collected randomly in southern Iraq, in AL-Nasiriya city, from AL Hussein teaching hospital. Number of samples is 100, 70 bladder cancer tissue and 30 controls benign tissue. Results of this study reveal that pD_L1 expression is positive in 45 of 70 sample. The study demonstrated pD_L1 expression is increased in high grade bladder cancer represent (44.19%), while in low grade (33.33%), pD_L1 expression high in T2 stage (41.03%), and in Ta(26.3%) T1(33.3%) T3 (50%) T4(100%). expression pD_L1 excessive in muscle invasive (42.86%), whilst the low expression for pD_L1 in the non-muscle invasive type (35.71) So pD_L1 can be used as a marker for assessment of bladder cancer aggressiveness. This study represents an important step because there are few of studies about this topic in Iraq; we are needing more studies to prove the function of this pD_L1 in biological behavior of bladder cancer. [ABSTRACT FROM AUTHOR]

The aim of this study is to determine the expression of the FGFR3protein in human bladder cancer, and significant correlations between these parameters and clinico pathologic variables like (grade and stage. the bladder cancer is one of the most common cancers worldwide. the incidence and mortality of this disease increased during last decades alarmingly in Iraq. The current study is designed to detect the role of FGFR3expression in bladder carcinoma as a possible marker for detecting the biological behavior of malignancy and its correlation with grade and non-muscle invasive bladder cancer NMIBC. for both diagnostic and prognostic purposes. The study focuses on a technique of immunohistochemistry for detection FGFR3expression in bladder cancer. The samples are collected randomly in southern Iraq, in AL-Nasiriya city, from AL Hussein teaching hospital. Number of samples is 100, 70 bladder cancer tissue and 30 controls benign tissue. Results of this study reveal that FGFR3expression is positive in 49 of 70 sample. The study demonstrated FGFR3 expression is increased in low grade bladder cancer represent (96.30%), while in high grade (53.49%), FGFR3expression high in Ta stage (100%), and in T1(100%) T2(51.28) T3 (50%) expression FGFR3excessive in non-muscle invasive (92.85%), whilst the low expression for FGFR3in the muscle invasive type (54.76%). So FGFR3can be used as a marker for assessment of bladder cancer aggressiveness. This study represents an important step because there are few of studies about this topic in Iraq; we are needing more studies to prove the function of this FGFR3in biological behavior of bladder cancer. [ABSTRACT FROM AUTHOR]

The aim of current study is to determine the expression of the CK14 protein in human bladder cancer, and demonstration of correlations between this parameter and clinical pathologic variables as (grade and stage of tumor), and also by using this marker can classify bladder cancer in to basal and luminal type. The current study is designed to detect the role of Ck14 expression in bladder carcinoma as a possible marker for detecting the biological behavior of malignancy and its correlation with grade and muscle invasiveness for both diagnostic and prognostic purposes. The study focuses on a technique of immunohistochemistry for detection Ck14 expression in bladder cancer. The samples are collected randomly in southern Iraq, in AL-Nasiriya city, from AL Hussein teaching hospital. Number of samples is (100) samples, (70) bladder cancer tissue and (30) controls benign tissue. Results of this study reveal that Ck14 expression is positive in 45 out of 70 sample. The study present showed Ck14 expression is increased in high grade bladder cancer represent (86.0%), while in low grade (29.6%), CK14expression high in advanced tumour stages T3, T4 percentage (100%) and T2 stage (87.2%), expression of CK14 low in early tumor stages Ta (26.3) T1(33.3). expression CK14 excessive in muscle invasive (97.6%), whereas the expression CK14 is low in non-muscle invasive type (28.6%). So Ck14 can be used as a marker for assessment of bladder cancer aggressiveness. This study represents an important step because there is absence of studies about this topic in Iraq. [ABSTRACT FROM AUTHOR]

Benign prostatic hyperplasia (BPH) is an age-related enlargement of the prostate with urethral obstruction that predominantly affects the middle-aged and older male population, resulting in disruptive lower urinary tract symptoms (LUTS), thus creating a profound impact on an individual's quality of life. The development of LUTS may be linked to overexpression of oxytocin receptors (OXTR), resulting in increased baseline myogenic tone within the prostate. Thus, it is hypothesised that targeting OXTR using oxytocin receptor antagonists (atosiban, cligosiban, and β-Mercapto-β,β-cyclopentamethylenepropionyl1, O-Me-Tyr2, Orn8]-Oxytocin (ßMßßC)), may attenuate myogenic tone within the prostate. Organ bath and immunohistochemistry techniques were conducted on prostate tissue from young and older rats. Our contractility studies demonstrated that atosiban significantly decreased the frequency of spontaneous contractions within the prostate of young rats (**** p < 0.0001), and cligosiban (* p < 0.05), and ßMßßC (**** p < 0.0001) in older rats. Additionally, immunohistochemistry findings revealed that nuclear-specific OXTR was predominantly expressed within the epithelium of the prostate of both young (*** p < 0.001) and older rats (**** p < 0.0001). In conclusion, our findings indicate that oxytocin is a key modulator of prostate contractility, and targeting OXTR is a promising avenue in the development of novel BPH drugs. [ABSTRACT FROM AUTHOR]

The liver, as a crucial metabolic organ, undergoes significant pathological changes during the aging process, which can have a profound impact on overall health. To gain a comprehensive understanding of these alterations, we employed data-driven approaches, along with biochemical methods, histology, and immunohistochemistry techniques, to systematically investigate the effects of aging on the liver. Our study utilized a well-established rat aging model provided by the National Institute of Aging. Systems biology approaches were used to analyze genome-wide transcriptomics data from liver samples obtained from young (4–5 months old) and aging (20–21 months old) Fischer 344 rats. Our findings revealed pathological changes occurring in various essential biological processes in aging livers. These included mitochondrial dysfunction, increased oxidative/nitrative stress, decreased NAD + content, impaired amino acid and protein synthesis, heightened inflammation, disrupted lipid metabolism, enhanced apoptosis, senescence, and fibrosis. These results were validated using independent datasets from both human and rat aging studies. Furthermore, by employing co-expression network analysis, we identified novel driver genes responsible for liver aging, confirmed our findings in human aging subjects, and pointed out the cellular localization of the driver genes using single-cell RNA-sequencing human data. Our study led to the discovery and validation of a liver-specific gene, proprotein convertase subtilisin/kexin type 9 (PCSK9), as a potential therapeutic target for mitigating the pathological processes associated with aging in the liver. This finding envisions new possibilities for developing interventions aimed to improve liver health during the aging process. [ABSTRACT FROM AUTHOR]

Background: Pulmonary fibrosis (PF) is a terminal pathological change in a variety of lung diseases characterized by excessive deposition of extracellular matrix, for which effective treatment is lacking. Tangeretin (Tan), a flavonoid derived from citrus, has been shown to have a wide range of pharmacological effects. This study aimed to investigate the role and potential mechanisms of Tan on pulmonary fibrosis. Methods: A model of pulmonary fibrosis was established by administering bleomycin through tracheal drip, followed by administering Tan or pirfenidone through gavage. HE and Masson staining were employed to assess the extent of pulmonary fibrosis. Subsequently, Western blot, enzyme-linked immunosorbent assay (ELISA), RNA sequencing, and immunohistochemistry techniques were employed to uncover the protective mechanism of Tan in PF mice. Furthermore, A549 cells were stimulated with TGF-β1 to induce epithelialmesenchymal transition (EMT) and demonstrate the effectiveness of Tan in mitigating PF. Results: Tan significantly ameliorated bleomycin-induced pulmonary fibrosis, improved fibrotic pathological changes, and collagen deposition in the lungs, and reduced lung inflammation and oxidative stress. The KEGG pathway enrichment analysis revealed a higher number of enriched genes in the PI3K/ Akt pathway. Additionally, Tan can inhibit the EMT process related to pulmonary fibrosis. Conclusion: Taken together, the above research results indicate that Tan suppresses inflammation, oxidative stress, and EMT in BLM-induced pulmonary fibrosis via the PI3K/Akt pathway and is a potential agent for the treatment of pulmonary fibrosis. [ABSTRACT FROM AUTHOR]