Tissue Histology Research Articles

MiR-223 and Chromogranin A Affect Inflammatory Immune Cell Activation in Liver Metastasis of Neuroendocrine Neoplasms

Neuroendocrine neoplasms (NENs) are a diverse group originating from endocrine cells/their precursors in pancreas, small intestine, or lung. The key serum marker is chromogranin A (CgA). While commonly elevated in patients with NEN, its prognostic value is still under discussion. Secretion/posttranslational proteolytic cleavage of CgA results in multiple bioactive fragments, which are essential regulators of the cardiovascular and immune system. miR-223, regulator of Nrlp3 inflammasome and neutrophil activation, was recently found to have decreased in patients with NEN. We performed flow cytometry of circulating neutrophils in a patient cohort (n = 10) with NEN, microdissection and histology of tumor tissue. Subsequently, in vitro transfections using the well-established human pancreatic NEN cell line (BON), and co-culture experiments with primary macrophages and neutrophils were performed. Serum miR-223 in patients correlated with the expression of the neutrophil activation marker CD15 in circulating cells. Neutrophilic CD62L/CD63 showed good discrimination compared to healthy controls. Immune cell-derived miR-155, miR-193 and miR-223 colocalize with neutrophil in the extra-tumoral tissue alongside Nlrp3-associated caspase-1 activation. miR-223 knockdown in BON decreased the CgA intracellularly, increased in cellular granularity and caspase-1 activation. Plasmin inhibitor a2-aP reverted those effects. Western Blot showed fragmented CgA following miR-223 knockdown, which altered the inflammatory potential of neutrophils. Our data hence provide initial insights into an immunoregulatory mechanism via miR-223 and CgA in NEN cells, as regulation of miR-223 in NEN may affect tumor-associated inflammation.

Effect of Food Deprivation on Plasma Cortisol, Carbohydrate Metabolism, and Histomorphology in Clarias batrachus.

The nutritional status of fish is essential for its health, experimental studies, and aquaculture practices. The current study investigated the impact of food deprivation on biochemical parameters, histology of skin, gill, and kidney tissues, and ultrastructure of gills in Clarias batrachus. Fish were subjected to food deprivation for 2, 7, and 15 days resulting in (a) significant increase in plasma cortisol levels, (b) no significant changes in plasma osmolality and plasma glucose content, and (c) significant decrease in liver and muscle glycogen contents. A substantial damage was detected in skin, gill, and kidney tissues with histological alterations in a time-dependent manner. Skin tissue displayed melanomacrophage aggregation, excoriated epidermis and dermis. In gill tissue, epithelial lifting, edema, desquamation, deformed secondary lamellae, and lamellar hyperplasia were observed. Kidney tissue exhibited degenerated tubules, melanomacrophage aggregations, and shrunken renal tubule. Scanning electron microscopy revealed that food deprivation-induced marked presence of mucus, chloride cells, and pavement cells with well-defined microridges and microbridges following 2 days, opening of chloride cells was more prominent after 7 days, while more mucus secretion was observed after 15 days. After food deprivation, alterations in biochemical and histological parameters, and ultrastructural changes in target tissues reflect physiological and morphological disturbances in fish. The novelty of this study is that these parameters can be considered as biomarkers of feeding stress in fish and fish health and can provide important insights for better aquaculture practices.

Tissue histology and depuration of per- and polyfluoroalkyl substances (PFAS) from dairy cattle with lifetime exposures to PFAS-contaminated drinking water and feed

Plasma, milk and tissue samples were collected from 30 dairy cattle (0.4 to 8.9 years of age) with lifetime exposures to perfluoroalkyl substances (PFAS) removed from a PFAS-contaminated farm and provided PFAS-free feed and water. Twenty cattle were slaughtered 2 weeks after removal from the farm and tissues were collected for histological and residue analyses. Milk and/or plasma were collected from all remaining cattle at 2-week intervals and milk samples were collected daily but were analyzed at the same intervals as plasma samples. The remaining cattle were slaughtered 20 and 22 weeks after the initial set of 20 animals were slaughtered. While many incidental and normal background findings were noted on histological evaluation, no consistent histological finding was associated with PFAS exposure. Perfluoroalkyl carboxylic acids (PFCA) and perfluoro butane sulfonic acid (PFBS) were not generally detected in milk, plasma and tissues, but perfluoroalkyl sulfonic acids (PFSA) were quantifiable throughout the 22-week withdrawal period in most matrices. Estimated plasma half-lives of perfluorohexane sulfonic acid (PFHxS), perfluoroheptane sulfonic acid (PFHpS), linear perfluorooctane sulfonic acid (L-PFOS), perfluoro-3-methyl heptanesulfonate (3Me-PFOS) and perfluoro-6-methyl heptanesulfonate (6Me-PFOS) ranged from 4 to 10 weeks, but the estimates were associated with large confidence intervals. Across animal status (heifer, lactating, dry), natural log transformed (Ln) plasma residues of PFHxS and L-PFOS were generally well correlated with Ln-transformed PFHxS and L-PFOS residues in lung, muscle, liver and kidney (R2, 0.7572 to 0.9394) whereas the strongest relationships of Ln-transformed L-PFOS residues among tissues were between lung and liver, kidney and muscle (R2, 0.8287 to 0.9138).

Atractylenolide-III restrains cardiac fibrosis after myocardial infarction via suppression of the RhoA/ROCK1 and ERK1/2 pathway.

Cardiac fibrosis, a critical factor in myocardial remodeling post-myocardial infarction (MI), can advance heart failure progression. Atractylenolide III (ATL-III), derived from Atractylodes lancea, has recognized antioxidant and anti-inflammatory effects; however, its influence on cardiac fibrosis remains unclear. MI was induced in mice by permanent ligation of the left anterior descending (LAD) coronary artery, followed by 2weeks of ATL-III or dimethyl sulfoxide (DMSO) treatment. Cardiac fibrosis was assessed by echocardiography, tissue histology, and serum biomarkers of myocardial injury. In vitro, the effects of ATL-III on cardiac fibroblast (CF) proliferation and collagen deposition were evaluated using immunofluorescence, 5-Ethynyl-2'-deoxyuridine (EdU), and western blot techniques. Network pharmacology and molecular docking identified potential ATL-III targets. ATL-III treatment significantly improved cardiac function, as evidenced by increased ejection fraction (EF) and fractional shortening (FS) and reduced left ventricular dilation. Histological analysis revealed decreased fibrotic areas in ATL-III-treated mice, along with reduced expression of fibrosis markers α-SMA and Collagen I. ATL-III also alleviated oxidative stress by reducing reactive oxygen species (ROS) and malondialdehyde (MDA) levels while increasing superoxide dismutase (SOD) activity. Furthermore, ATL-III suppressed inflammation, decreasing TNF-α, IL-6, and IL-1β protein and mRNA levels. In vitro, ATL-III inhibited TGF-β1-induced CF proliferation, migration, and differentiation, reducing the expression of fibrotic markers. Mechanistically, ATL-III suppressed the RhoA/ROCK1 and ERK1/2 signaling pathways, as confirmed by molecular docking and pathway analysis. ATL-III demonstrates therapeutic potential in mitigating post-MI cardiac fibrosis by reducing oxidative stress, inflammation, and CF activation. These findings highlight ATL-III as a promising candidate for the treatment of cardiac fibrosis and associated heart failure.

Clinical and histomorphometric evaluation of the vagina following treatment with CO2 laser, radiofrequency, and promestriene for genitourinary syndrome of menopause in breast cancer survivors on adjuvant therapy.

To perform clinical and histomorphometric evaluations of the vagina before and after treatment for genitourinary syndrome of menopause with CO2 laser, fractional radiofrequency, and promestriene in breast cancer survivors using adjuvant endocrine therapy. The study included women experiencing moderate to severe genitourinary syndrome of menopause. Following pre- and post-treatment protocols, participants graded their symptoms on a visual analog scale and underwent a gynecological examination to evaluate the Vaginal Health Index and obtain a vaginal biopsy. Subsequently, they were randomized to laser, radiofrequency, and promestriene groups. The energy groups underwent three consecutive monthly outpatient vulvovaginal treatments, while the control group received promestriene treatment for four months, followed by a post-treatment evaluation. Sixty-two women completed the study protocol (21 laser, 20 radiofrequency, and 21 promestriene). While histological differences (vaginal thickness [p=0.002] and number of stromal papillae [p=0.004]) were observed between the pretreatment samples of tamoxifen and anastrozole users, the symptoms did not differ between them. A decrease in symptom intensity (p<0.05) and an improvement in the Vaginal Health Index (p<0.001) were observed post-treatment, regardless of the type of adjuvant endocrine therapy used. Most pretreatment vaginal samples did not indicate histological atrophy, and no significant histological differences were observed after treatment. No clinical or histological damage was observed. CO2 laser and radiofrequency therapies could be considered alternative treatments for genitourinary syndrome of menopause in breast cancer survivors receiving adjuvant therapy. These treatments promoted significant improvements comparable to those delivered by promestriene, without histological or clinical tissue damage. NCT04081805.

Ameliorating potential of clove oil on histological tissue damage induced by TCDD in lung of Wister albino Rats

Ameliorating potential of clove oil on histological tissue damage induced by TCDD in lung of Wister albino Rats

Histopathological alteration in Zebrafish: Unravelling the Effects of Long-Term Copper Oxide Nanoparticle Exposure

Copper Oxide nanoparticles (CuO-NPs) are widely used and they build up in the aquatic environment and can be harmful to aquatic life. Aquatic creatures are affected by CuO NPs, yet the consequences of these particles have not been well explored despite contentious toxicological results. Therefore, this study aimed to investigate the effects of chronic exposure to CuO-NPs on adult zebrafish. The study was carried out by conducting physicochemical characterization of commercially procured CuO-NPs using UV/Vis, ICP-OES spectroscopy, Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), and Fourier Transform Infrared Spectroscopy(FTIR). Following characterization, adult zebrafish were chronically exposed to 0.5, 1, and 3 mg/l of CuO-NPs. Characterization revealed a heterogeneous population of nanoparticles in the size ranges of 10–50 nm. Oxidative stress indicators and functional markers were studied in addition to tissue histology. Chronic exposure to adult Fish at concentrations of 1 and 3 mg/l exhibited increased levels of stress compared to lower concentrations of 0.5 mg/l. Observations from histology showed that the extent of tissue damage and injuries increased with the concentration of CuO-NPs. In conclusion, chronic exposure to ≤50nm-sized CuO-NPs exhibited toxic repercussions in adult zebrafish.

Utilization of breast magnetic resonance imaging in the pre-operative setting and its ability to better predict local staging in non-metastatic breast cancer patients

Background: Breast cancer can be difficult to stage using ultrasound (USS) and mammography (MMG) alone. The use of MRI in the pre-operative setting has been increasingly incorporated to aid in treatment planning. However, the indications for breast magnetic resonance imaging (MRI) are not universally agreed upon, and its direct impact on patient care requires further research. This paper aims to assess pre-operative non-metastatic breast cancer patients to determine whether breast MRI led to changes in management and to evaluate its accuracy in predicting tissue staging when compared to ultrasound and mammography. Method: This is a retrospective study of 44 patients diagnosed with non-metastatic breast cancer who underwent upfront surgery and received pre-operative breast MRI between March 1, 2022, and March 1, 2024. Patient demographics, pre-operative imaging (including USS, MMG, and MRI), and final tissue histology were reviewed. The findings from breast MRI and its impact on pre-operative planning, as well as its concordance with final tissue histological staging, were assessed. Results: Of the 44 patients who underwent pre-operative breast MRI, 12 experienced a change in management, and 11 required additional investigations. This was largely due to an upstaging of the disease, leading to mastectomy instead of the previously planned breast-conserving surgery. Furthermore, breast MRI demonstrated better concordance with final histological staging at 69.4% compared to USS/MMG alone at 52.7%. Conclusion: Breast MRI is a valuable tool in pre-operative planning for newly diagnosed breast cancer patients. It can significantly affect surgical management in a substantial proportion of patients and has been shown to be more accurate in estimating disease burden.

The significance of the immunohistochemical method in diagnosing atypical endometrial polyps as manifestations of localized hyperplasia

The article explores the role of immunohistochemical methods in diagnosing atypical endometrial polyps as manifestations of localized hyperplasia. Endometrial polyps are a widespread pathology among women of reproductive age, frequently accompanied by hyperplastic tissue changes. The aim of the work was to investigate the expression of diagnostic markers in endometrial polyps, which may be manifestations of local endometrial hyperplasia, with the aim of improving differential diagnosis in their atypia.. The research material consisted of histological samples from 37 patients aged 38 to 52 with abnormal uterine bleeding, treated at a clinical hospital in Dnipro. The methods included histological tissue analysis, immunohistochemical studies using a panel of antibodies, and statistical data analysis. Antibodies targeting markers such as ER, PR, PAX-2, PAX-8, PTEN, Ki-67, p16, and p53 were used to evaluate proliferative activity and the degree of tissue atypia. The results showed that polyps associated with atypical hyperplasia are characterized by complex morphological changes, including abnormal glandular architecture, stromal atypia, and increased mitotic activity in both glandular and stromal components. Epithelial differentiation of tubal, mucinous, or mixed types was frequently observed. More than half of the cases exhibited partial atypical changes, while diffuse atypia was noted in fewer instances. The control group of glandular-fibrous polyps with hyperplasia without atypia demonstrated low mitotic activity and no cellular atypia. Immunohistochemical studies confirmed the high sensitivity of markers in identifying pathological chan­ges, particularly in diagnostically challenging cases. The conclusions emphasize that immunohistochemistry is an effec­tive tool for detecting endometrial atypia, and the diagnosis of atypical polyps should be based on a comprehensive analysis of morphological and molecular changes. Polyps associated with hyperplasia and atypia require further observation due to the risk of recurrence and potential malignant transformation. Utilizing markers such as PAX-2, PTEN, and Ki-67 enhances diagnostic accuracy. Additional studies are needed to develop more reliable risk stratification models.

Mesenchymal stromal cells reduce inflammation and improve lung function in a mouse model of cystic fibrosis lung disease

Mesenchymal stromal cells (MSCs) are multipotent adult stem cells which possess immunomodulatory and repair capabilities. In this study, we investigated whether MSC therapy could modulate inflammation and lung damage in the lungs of Scnn1b-transgenic mice overexpressing the β-subunit of the epithelial sodium channel (β-ENaC), a model with features of Cystic Fibrosis lung disease. Human bone marrow derived MSC cells were intravenously delivered to mice, prior to collection of bronchoalveolar lavage (BALF) and tissue. BALF analysis revealed a significant reduction in inflammatory cells after MSC administration, with both monocytic cells and neutrophils significantly reduced. Pro-inflammatory cytokines keratinocyte-derived chemokine (KC) and osteopontin were also significantly reduced. Histological tissue analysis revealed a reduction in emphysema in Scnn1b-TG mice treated with MSCs and consistent with these findings, improvements in lung function after MSC therapy were observed. Furthermore, MSCs enhanced Ki67 staining in alveolar cells, which may indicate regeneration of the destroyed parenchyma. Mechanistically, restoration of peroxisome proliferator-activated receptor-γ (PPARγ) expression and its transcriptional program were identified after MSC treatment. Our data demonstrate that MSC therapy can reduce inflammation, damage, and lung function decline in the chronically inflamed lung of Scnn1b-Tg mice, suggesting that MSCs may provide an effective tool in the treatment of muco-obstructive diseases such as cystic fibrosis.

Early detection of bacterial pneumonia by characteristic induced odor signatures

IntroductionThe ability to detect pathogenic bacteria before the onsets of severe respiratory symptoms and to differentiate bacterial infection allows to improve patient-tailored treatment leading to a significant reduction in illness severity, comorbidity as well as antibiotic resistance. As such, this study refines the application of the non-invasive Secondary Electrospray Ionization-High Resolution Mass Spectrometry (SESI-HRMS) methodology for real-time and early detection of human respiratory bacterial pathogens in the respiratory tract of a mouse infection model.MethodsA real-time analysis of changes in volatile metabolites excreted by mice undergoing a lung infection by Staphylococcus aureus or Streptococcus pneumoniae were evaluated using a SESI-HRMS instrument. The infection status was confirmed using classical CFU enumeration and tissue histology. The detected VOCs were analyzed using a pre- and post-processing algorithm along with ANOVA and RASCA statistical evaluation methods.ResultsCharacteristic changes in the VOCs emitted from the mice were detected as early as 4–6 h post-inoculation. Additionally, by using each mouse as its own baseline, we mimicked the inherent variation within biological organism and reported significant variations in 25 volatile organic compounds (VOCs) during the course of a lung bacterial infection.Conclusionthe non-invasive SESI-HRMS enables real-time detection of infection specific VOCs. However, further refinement of this technology is necessary to improve clinical patient management, treatment, and facilitate decisions regarding antibiotic use due to early infection detection.

Ecological risk assessment of sewage sludge as soil amendment: lethal and sublethal effects in Eisenia fetida.

Sewage sludge applications as soil amendment call for a proper ecological risk assessment due to unexpected delivery of toxic chemicals and materials. Standardized acute toxicity assays have proven to provide limited information in terms of potential hazard for soil organisms. Here, sublethal endpoints as physiological and tissue alterations were proposed as suitable tools for sewage sludge ecological risk assessment. Acute toxicity assay using earthworm Eisenia fetida (OECD n.207) was performed on sewage sludge samples from two wastewater treatment plants (A and B), and mortality, body weight, body morphology, and histology of intestinal cavity were evaluated at the end of the exposure period. Sewage sludge dilutions (1:50 and 1:100) from plant B caused a low percentage of mortality (5%), while none was observed in worms exposed to plant A. However, all specimens showed external body alterations as vesicles, swellings, constrictions, and discoloration. Histological sections highlight damages to epidermis, circular and longitudinal muscles, and chloragogenic tissue regardless of sewage sludge origin. Our findings confirm that standardized end points of acute toxicity are not sensitive enough in worms exposed to sewage sludges, while sublethal endpoints as body morphology and tissue histology are thus more suitable and appropriate endpoints for assessing potential detrimental outcomes and ecological risk for soil biota.

Rapid Stiffness Mapping in Soft Biologic Tissues With Micrometer Resolution Using Optical Multifrequency Time-Harmonic Elastography.

Rapid mapping of the mechanical properties of soft biological tissues from light microscopy to macroscopic imaging can transform fundamental biophysical research by providing clinical biomarkers to complement in vivo elastography. This work introduces superfast optical multifrequency time-harmonic elastography (OMTHE) to remotely encode surface and subsurface shear wave fields for generating maps of tissue stiffness with unprecedented detail resolution. OMTHE rigorously exploits the space-time propagation characteristics of multifrequency time-harmonic waves to address current limitations of biomechanical imaging and elastography. Key solutions are presented for stimulation, wave decoding, and stiffness reconstruction of shear waves at multiple harmonic frequencies, all tuned to provide consistent stiffness values across resolutions from microns to millimeters. OMTHE's versatility is demonstrated by simulations, phantoms, Bacillus subtilis biofilms, zebrafish embryos and adult zebrafish, reflecting the diversity of biological systems from a mechanics perspective. By zooming in on stiffness details from coarse to finer scales, OMTHE has the potential to advance mechanobiology and offers a way to perform biomechanics-based tissue histology that consistently matches in vivo time-harmonic elastography in patients.

Human placental stem cell-based therapies for prevention of abdominal adhesions: A prospective randomized preclinical trial.

Abdominal adhesions are networks of fibrotic tissues that form between organs postoperatively. Current prophylactic strategies do not reproducibly prevent adhesive small bowel obstruction across the entire abdomen. Human placental-derived stem cells produce an anti-inflammatory secretome that has been applied to multiple fibrosing diseases. The purpose of this project is to test human placental stem cell (hPSC)-based therapies for prevention of abdominal adhesions in a clinically relevant rat model. Fifty-four (n = 54, n = 6/group) male Sprague-Dawley rats (250-350 g) underwent model creation and treatment randomization under anesthesia. Experimental groups included human placental-derived stem cells (hPSC, 5 × 106 cells/10 mL Plasmalyte A), human placental-derived stem cells in a hyaluronic acid (HA-Mal-hPSC) hydrogel, the human placental-derived stem cell secretome from conditioned media in 10 mL Plasmalyte A, human placental-derived stem cells' conditioned media in a hyaluronic acid (HA-Mal-CM) hydrogel, Plasmalyte A (media alone, 10 mL), hyaluronic acid hydrogel alone (HA-Mal), Seprafilm (Baxter, Deerfield, IL), and the control groups, model with no treatment (MNT) and sham animals. Treatments were administered intraperitoneally, and the study period was 14 days postoperation. Adhesions were scored at necropsy and analyzed as the difference between means of an index statistic (Animal Index Score) versus MNT. Underlying molecular mechanisms were explored by functional genomic analysis and histology of peritoneal tissues. Hyaluronic acid hydrogel alone, HA-Mal-CM hydrogel, and Seprafilm significantly reduced the overall appearance of abdominal adhesions by mean Animal Index Score at 14 days versus MNT. Human placental stem cell, HA-Mal-hPSC hydrogel, HA-Mal-CM hydrogel, HA-Mal hydrogel alone, and Seprafilm significantly reduced the collagen content of injured peritoneal tissues. Human placental stem cell and HA-Mal-hPSC hydrogel suppressed expression of the most profibrotic genes. Conditioned media, HA-Mal hydrogel alone, and media alone significantly altered the expression of proteins associated with peritoneal fibrotic pathways. Human placental stem cell-based therapies reduce abdominal adhesions in a prospective randomized preclinical trial. This effect is supported by suppression of profibrotic genomic and proteomic pathways.

Combination of Deep and Statistical Features of the Tissue of Pathology Images to Classify and Diagnose the Degree of Malignancy of Prostate Cancer.

Prostate cancer is one of the most prevalent male-specific diseases, where early and accurate diagnosis is essential for effective treatment and preventing disease progression. Assessing disease severity involves analyzing histological tissue samples, which are graded from 1 (healthy) to 5 (severely malignant) based on pathological features. However, traditional manual grading is labor-intensive and prone to variability. This study addresses the challenge of automating prostate cancer classification by proposing a novel histological grade analysis approach. The method integrates the gray-level co-occurrence matrix (GLCM) for extracting texture features with Haar wavelet modification to enhance feature quality. A convolutional neural network (CNN) is then employed for robust classification. The proposed method was evaluated using statistical and performance metrics, achieving an average accuracy of 97.3%, a precision of 98%, and an AUC of 0.95. These results underscore the effectiveness of the approach in accurately categorizing prostate tissue grades. This study demonstrates the potential of automated classification methods to support pathologists, enhance diagnostic precision, and improve clinical outcomes in prostate cancer care.

Endoscopic Ultrasound-Guided Pancreatic Tissue Sampling: Lesion Assessment, Needles, and Techniques.

Endoscopic ultrasound (EUS)-guided tissue sampling includes the techniques of fine needle aspiration (FNA) and fine needle biopsy (FNB), and both procedures have revolutionized specimen collection from the gastrointestinal tract, especially from remote/inaccessible organs. EUS-FNB has replaced FNA as the procedure of choice for tissue acquisition in solid pancreatic lesions (SPLs) across various society guidelines. FNB specimens provide a larger histological tissue core (preserving tissue architecture) with fewer needle passes, and this is extremely relevant in today's era of precision and personalized molecular medicine. Innovations in needle tip design are constantly under development to maximize diagnostic accuracy by enhancing histological sampling capabilities. But, apart from the basic framework of the needle, various other factors play a role that influence diagnostic outcomes, namely, sampling techniques (fanning, aspiration or suction, and number of passes), collection methods, on-site evaluation (rapid, macroscopic, or visual), and specimen processing. The choice taken depends strongly on the endoscopist's preference, available resources at the disposal, and procedure objectives. Hence, in this review, we explicate in detail the concepts and available literature at our disposal on the topic of EUS-guided pancreatic tissue sampling to best guide any practicing gastroenterologist/endoscopist in a not-to-ideal set-up, which EUS-guided tissue acquisition technique is the "best" for their case to augment their diagnostic outcomes.

Protective effect of Huanglian Pingwei San on DSS-induced ulcerative colitis in mice through amelioration of the inflammatory response and oxidative stress.

Ulcerative colitis (UC) results in the breakdown of the mucosal barrier caused by persistent inflammation and oxidative stress. Huanglian Pingwei San (HLPWS) is a commonly prescribed traditional Chinese medicine for treating colitis, but the precise mechanism remains unclear. The aim of this study was to systematically investigate the protective effect of HLPWS on UC mice and to elucidate the underlying mechanisms involved. UC mouse model was established in C57BL/6 mice via 2.25% dextran sulfate sodium (DSS). The chemical composition of HLPWS was examined through UPLC/MS Q-TOF analysis. The efficacy of HLPWS in treating UC was assessed. A TUNEL assay was used to detect apoptotic cells. An ELISA was used to evaluate the levels of inflammatory cytokines in colon tissues and serum. The percentages of Treg and Th17 cells were measured via flow cytometry. The protein expression in the colonic tissue was validated via immunohistochemistry (IHC) and Western blotting. HLPWS significantly improved UC symptoms and colon tissue histology in mice. The structure and function of the intestinal barrier were restored by HLPWS treatment, as shown by increased DAO content, reduced levels of FITC-dextran, and increased protein expression of ZO-1, occludin, claudin, and MUC2. HLPWS dose-dependently decreased the number of apoptotic cells by inhibiting P53, P21, P27, cleaved caspase 3, and p-H2AX expression. HLPWS also reduced abnormal oxidative stress by reducing Keap1 expression and increasing Nrf2 and HO-1 levels. Furthermore, HLPWS rebalanced the Treg/Th17 ratio to alleviated inflammatory reactions in UC mice. These findings suggest that HLPWS alleviated colonic intestinal barrier dysfunction in UC mice by reducing oxidative stress and restoring immune balance. This study underscores the potential therapeutic benefits of HLPWS and highlights its potential as a future pharmaceutical candidate for UC treatment.

Modified human mesenchymal stromal/stem cells restore cortical excitability after focal ischemic stroke in rats.

Allogeneic modified bone marrow-derived mesenchymal stromal/stem cells (hMSC-SB623 cells) are in clinical development for the treatment of chronic motor deficits after traumatic brain injury and cerebral ischemic stroke. However, their exact mechanisms of action remain unclear. Here, we investigated the effects of this cell therapy on cortical network excitability, brain tissue and peripheral blood at a chronic stage after ischemic stroke in a rat model. One month after focal cortical ischemic stroke, hMSC-SB623 cells or the vehicle solution were injected into the peri-stroke cortex. Starting one-week post-treatment, cortical excitability was assessed ex vivo. hMSC-SB623 cell transplants reduced stroke-induced cortical hyperexcitability, restoring cortical excitability to control levels. Histology of brain tissue revealed an increase of factors relevant to neuroregeneration and synaptic and cellular plasticity. Whole-blood RNA sequencing and serum protein analyses showed that intracortical hMSC-SB623 cell transplantation reversed effects of stroke on peripheral blood factors known to be involved in stroke pathophysiology. Our findings demonstrate that intra-cortical transplants of hMSC-SB623 cells correct stroke-induced circuit disruptions even at the chronic stage, suggesting broad utility as a therapeutic for neurological conditions with network hyperexcitability. Additionally, the transplanted cells exert far-reaching immunomodulatory effects whose therapeutic impact remains to be explored.

Fisetin ameliorates polycystic ovary syndrome in rats via a mechanistic modulation of AMP-activated protein kinase and SIRT1 molecular pathway.

Fisetin, a polyphenolic flavonoid, exhibits numerous pharmacological activities against metabolic syndromes. The present research aims to explore the therapeutic efficacy of fisetin in experimental polycystic ovary syndrome (PCOS). Female Sprague-Dawley rats were administered mifepristone (20mg/kg/day) to induce PCOS. PCOS rats were treated with fisetin (20mg/kg and 40mg/kg) and further compared with metformin HCl, the conventional drug for PCOS. The mechanism of fisetin was explored using dorsomorphin (an AMPK inhibitor). Then, rats were sacrificed for further analysis of biochemical and histological parameters. PCOS rats exhibited irregular estrous cycles, increased serum testosterone (4.72 ± 0.139ng/ml), estradiol (750.2 ± 16.56pg/ml), LH (30.33 ± 1.563 mIU/ml), HOMA-IR (1.115 ± 0.049), TNF-α (86.59 ± 3.93pg/ml), IL-6 (55.34 ± 4.432pg/ml), and TBARS (3.867 ± 0.193µmol/mg) along with declined progesterone (11.67 ± 1.54ng/ml), FSH (13.33 ± 1.256 mIU/ml), GSH (33.47 ± 1.348µmol/mg) levels, and SOD (2.163 ± 0.298 U/mg) activity as compared to normal control group. Fisetin high dose significantly lowers testosterone (3.014 ± 0.234ng/ml), estradiol (533.7 ± 15.39pg/ml), LH (16.67 ± 1.62 mIU/ml), HOMA-IR (0.339 ± 0.20), TNF-α (46.02 ± 2.66pg/ml), IL-6 (31.77 ± 3.47pg/ml), and TBARS (1.747 ± 0.185µmol/mg) and enhances progesterone (33.17 ± 1.447ng/ml), FSH (27.17 ± 1.42 mIU/ml), GSH (60.35 ± 1.1.102µmol/mg) levels, and SOD (4.513 ± 0.607 U/mg) activity. The histology of ovarian tissues shows a significant increase in cystic follicles in PCOS rats compared with the normal control group. These alterations were attenuated with fisetin treatment. Administration of dorsomorphin with fisetin can reverse the beneficial effects of fisetin in PCOS rats. Altogether, these present findings highlight the potential of fisetin as a promising therapeutic intervention for the management of PCOS by modulating AMPK/SIRT1 signaling in rats.

Expression of the Hopeful Therapeutic Target CD15 in Women with Breast Cancer

Background: CD15 is emerging as a recent prognostic marker with potential as a future therapeutic target in human malignancies. Its biological role in human cancer involves facilitating tumor cell adhesion through interaction with E-, L-, and P-selectins. This interaction allows adhesion to endothelial cells, enhancing metastatic potential. Objective: to study CD15 expression and its relationship to some pathologic parameters to provide the first Iraqi data-driven analysis of this growing concept in women with breast cancer. Subjects and Methods: A retrospective study was carried out utilizing tissue samples obtained from 85 patients with invasive breast cancer. Materials included one formalin-fixed paraffin-embedded tissue block, one hematoxylin and eosin-stained histological section, and two immunohistochemical-stained ER and HER2/NEU sections for each patient. The samples were obtained through core needle biopsy under ultrasound guidance. Data review and immunohistochemical staining with CD15 were conducted, followed by statistical analysis using the chi-square test to explore correlations. Results: CD15 expression was observed in 55% of breast cancer samples. There was a highly significant statistical relationship between CD15 expression and the presence of axillary lymph node metastasis (proven by fine needle aspiration cytology). However, there was no significant statistical relationship between CD15 expression and the histological grade, ER receptor, HER2-NEU status, and tissue calcification. Conclusion: CD15 expression was detected in breast cancer cells, with a highly significant association observed between CD15 expression and axillary lymph node metastasis at presentation. CD15 could serve as a prognostic marker as association with axillary lymph nodes reflects negative impact on prognosis and at the same time a promising therapeutic target for patients with breast cancer as blockage of CD15 antigen’s function will result in reduction of the metastatic potential and it could be an immunotherapeutic target.