Histomorphological Analysis Research Articles

Deficiency of FABP7 Triggers Premature Neural Differentiation in Idiopathic Normocephalic Autism Organoids

AbstractAutism spectrum disorder (ASD), which is caused by heterogeneous genetic and environmental factors, is characterized by diverse clinical phenotypes linked to distinct pathological mechanisms. ASD individuals with a shared clinical phenotype might contribute to revealing the molecular mechanism underlying ASD progression. Here, it is generated induced pluripotent stem cell (iPSC)‐derived cerebral organoids from normocephalic individuals with ASD in a prospective birth cohort with a shared clinical diagnosis. Multiple cell lines and time series scRNA‐seq combined with a histomorphological analysis revealed premature neural differentiation of neural stem cells (NSCs) and decreased expression of Fatty acid binding protein 7 (FABP7) in ASD organoids. It is subsequently revealed alterations in the phosphorylation levels of Mitogen‐Activated Protein Kinase Kinase 1/2 (MEK1/2), which are downstream of FABP7, and the regulation of the FABP7/MEK pathway reversed improper neural differentiation in the ASD organoids. Moreover, both Fabp7‐knockdown and MEK2‐overexpressing mice exhibited repetitive stereotyped behaviors and social defects relevant to autism. This study reveals the role of the FABP7/MEK pathway in abnormal NSC differentiation in normocephalic individuals with ASD, which might provide a promising therapeutic target for ASD treatment.

Optimising growth performance, nutrients digestibility, immunity and gut health in broilers through ginger-derived phyto-protease enzyme (zingibain) supplementation

This study investigated the impact of ginger-derived phyto-protease enzyme on growth performance, immunity, gut health and nutrient digestibility in broilers. A total of 360 day-old broiler chickens were randomly assigned to four treatments with six replicates each: a control group (T1) and groups supplemented with 50 g/ton (T2), 100 g/ton (T3) and 150 g/ton (T4) of the enzyme. Over a 35-day period, birds were provided with free access to feed and water under controlled conditions. The highest feed intake was observed in T3 during the second week, while T4 showed the lowest intake overall. Body weight gain was significantly highest in T4, demonstrating enhanced growth performance. Feed conversion ratio (FCR) was lowest in T4, indicating improved feed efficiency. Carcase quality analysis revealed higher dressing percentages and organ weights (bursa, spleen and thymus) in T4 compared to the control. Gut pH levels were significantly lower in T3 and T4 across different segments of the gastrointestinal tract, enhancing nutrient absorption. Antibody titres against Newcastle disease virus were significantly higher in T4 on days 21 and 35, indicating improved immunity. Histomorphological analysis of the intestines showed increased villus height and width in T4, with reduced crypt depth, further supporting enhanced nutrient absorption. These findings suggest that the highest inclusion rate of ginger-derived phyto-protease enzyme (150 g/ton) optimally enhances growth performance, immunity, gut health and nutrient digestibility in broilers. This study highlights the potential benefits of incorporating ginger-derived phyto-protease in poultry diets to improve overall health and performance.

BCL11A expression worsens the prognosis of DLBCL and its co-expression with C-MYC predicts poor survival

Non-Hodgkin's lymphoma (NHL) is a significant global malignancy, with diffuse large B cell lymphoma (DLBCL) being the most prevalent subtype, accounting for 25-50% of newly diagnosed cases in China. Despite a 60% survival rate achieved with R-CHOP regiment for DLBCL, approximately 40% of patients experience relapse or develop resistance to treatment. While the oncogenic transcription factor B-cell chronic lymphocytic leukaemia/lymphoma 11A (BCL11A) has been implicated in various tumors, its specific role in DLBCL remains unclear. In this study, we conducted retrospective histomorphological and immunophenotypic analyses on paraffin sample tissues and collected fresh tissue samples for protein and mRNA analyses to investigate the relationship between BCL11A and DLBCL. Additionally, we classified DLBCL into subtypes based on cells of origin (COO) and examined the expressions of BCL11A, C-MYC, P53 and other protein expressions to better understand the factors contributing to poor clinical outcomes in DLBCL. Our findings revealed elevated BCL11A expression in DLBCL, with increased expression associated with worse prognosis and higher C-MYC expression. Patients exhibiting co-expression of C-MYC and BCL11A had significantly lower survival rates compared to those with singular expression. Furthermore, BCL11A protein expression levels demonstrated significant associations with P53 and C-MYC protein expression levels in the Germinal Center B-cell-like (GCB) subtype. These findings suggest that BCL11A may serve as a potential prognostic marker and therapeutic target for DLBCL.

Lycium barbarum polysaccharide alleviates ferroptosis in Sertoli cells through NRF2/SLC7A11/GPX4 pathway and ameliorates DEHP-induced male reproductive damage in mice

Di-(2-ethylhexyl)phthalate (DEHP) is a common plasticizer that has been shown to significantly negatively affect male reproductive health. On the other hand, Lycium barbarum polysaccharide (LBP) has been shown to improve reproductive function. Therefore, we hypothesized that LBP may ameliorate DEHP-induced male reproductive damage. Herein, we found that LBP could alleviate DEHP-induced testicular damage and sperm abnormalities. Furthermore, histomorphological analysis of mice testis revealed that LBP primarily ameliorated the DEHP-induced male reproductive damage by targeting Sertoli cells. Moreover, the detection of the function-related genes of Sertoli cells confirmed this finding. The serum of mice in the Control, DEHP alone, and DEHP+LBP groups was analyzed using non-targeted metabolomics to further elucidate the mechanism of action of LBP in improving DEHP-induced male reproductive damage. According to the results, the differential metabolites were mainly enriched in the glutamate metabolism pathway, implying that LBP may alleviate the ferroptosis-related DEHP-induced testicular injury. Related ferroptosis markers were also found in mice testis. These findings collectively suggest that LBP may ameliorate DEHP-induced testicular injury via alleviating ferroptosis in Sertoli cells. To clarify the specific mechanism, we constructed a cell model in vitro by treating TM4 cells (the Sertoli cell line) with LBP and MEHP (the in vivo DEHP metabolite). Our findings revealed that LBP can improve the function of DEHP-affected Sertoli cells. Furthermore, the analysis of lipid peroxidation, Fe2+ content, and ferroptosis-related protein expressions demonstrated that LBP could ameliorate MEHP-induced ferroptosis in TM4 cells. To clarify the specific mechanism, glutamate metabolism-related proteins involved in the ferroptosis pathway were detected. According to the results, there were significant changes in the expression of NRF2, SLC7A11 and GPX4 proteins, which are involved in the ferroptosis glutamate metabolism pathway. Furthermore, supplementation of NRF2, SLC7A11, and GPX4 inhibitors (ML385, Erastin, and RSL3, respectively) blocked the therapeutic effect of LBP in alleviating MEHP-induced ferroptosis in TM4 cells, implying that LBP could also ameliorate MEHP-induced ferroptosis via the NRF2/SLC7A11/GPX4 pathway. In summary, these findings show that LBP can alleviate DEHP/MEHP-induced ferroptosis through the NRF2/SLC7A11/GPX4 pathway, ameliorating Sertoli cell dysfunction and improving the DEHP-induced male reproductive damage. Therefore, the clinical administration of LBP could be an effective strategy for preventing DEHP-induced male reproductive injury.

Zuogui Wan modulates macrophage polarization and promotes osteogenic differentiation through regulation of CD51-positive bone marrow mesenchymal stem cells

Background Zuogui Wan (ZGW) is a traditional herbal formula used to treat chronic kidney and bone diseases. Previous research has shown that ZGW slows down the aging process of bone marrow mesenchymal stem cells (BMSCs) and improves bone metabolism. However, its role in treating postmenopausal osteoporosis (PMOP) has not yet been fully investigated. Therefore, we investigated the therapeutic effects of ZGW and its potential mechanisms in an ovariectomy (OVX)-induced osteoporosis rat model. Results We observed significant improvements in bone loss and the osteoporotic phenotype in OVX rats treated with ZGW. These findings were confirmed with micro-computed tomography (micro-CT) and histomorphological analysis. We also discovered that ZGW reversed the macrophage imbalance, which in turn inhibited osteoclast differentiation and bone resorption. Furthermore, RNA-Seq results revealed the active expression of CD51 in BMSCs before and after ZGW therapy, which is associated with macrophage polarization and osteoblastic differentiation. The results also showed that ZGW decreased CD51 + BMSCs levels, which is closely related to the inhibition of osteoblast differentiation and promotion of osteoclast resorption. Conclusions Our study demonstrated that ZGW may improve postmenopausal osteoporosis by restoring macrophage polarization and down-regulating CD51 + BMSCs. In addition, ZGW promoted osteoblast formation and inhibited osteoclast resorption.

Prognosis prediction via histological evaluation of cellular heterogeneity in glioblastoma

Glioblastomas (GBMs) are the most aggressive types of central nervous system tumors. Although certain genomic alterations have been identified as prognostic biomarkers of GBMs, the histomorphological features that predict their prognosis remain elusive. In this study, following an integrative diagnosis of 227 GBMs based on the 2021 World Health Organization classification system, the cases were histologically fractionated by cellular variations and abundance to evaluate the relationship between cellular heterogeneity and prognosis in combination with O-6-methylguanine-DNA methyltransferase gene promoter methylation (mMGMTp) status. GBMs comprised four major cell types: astrocytic, pleomorphic, gemistocytic, and rhabdoid cells. t-distributed stochastic neighbor embedding analysis using the histological abundance of heterogeneous cell types identified two distinct groups with significantly different prognoses. In individual cell component analysis, the abundance of gemistocytes showed a significantly favorable prognosis but confounding to mMGMTp status. Conversely, the abundance of epithelioid cells was correlated with the unfavorable prognosis. Linear model analysis showed the favorable prognostic utility of quantifying gemistocytic and epithelioid cells, independent of mMGMTp. The evaluation of GBM cell histomorphological heterogeneity is more effective for prognosis prediction in combination with mMGMTp analysis, indicating that histomorphological analysis is a practical and useful prognostication tool in an integrative diagnosis of GBMs.

Histomorphological effects of ß,Ɛ-Carotene-3,3’-diol on the liver following indomethacin-induced hepatotoxicity in adult male Wistar rats

Medicinal plants are globally used as food and in the management of different ailments based on their peculiar phytochemical contents. This study aimed to evaluate the histomorphological effects of ß,Ɛ-Carotene-3,3’-diol on the liver following indomethacin-induced hepatotoxicity in adult male Wistar rats. This is to determine the ameliorative effect of graded doses of ß,Ɛ-Carotene-3,3’-diol. A total of twenty-five adult Wistar rats were randomly divided into five groups (n=5). Liver injury was induced by oral administration of 20 mg/kg b.w of indomethacin after 24 hours fast to animals in groups B-E. Twenty four hours after the induction, animals in groups C-E were given different doses of ß,Ɛ-Carotene-3,3’-diol. At the end of the experimentation, the animals were sacrificed and the livers were collected and fixed for histopathological and histomorphological analyses. The results showed that indomethacin increased the activities of AST, ALT and ALP and also induced histopathological changes. Treatment with ß,Ɛ-Carotene-3,3’-diol was able to attenuate liver injury caused by indomethacin in a dose dependent manner as evidenced in the histoarchitectural changes. This was also corroborated by the histomorphometric analyses. In conclusion, the results suggested the positive influence of ß,Ɛ-Carotene-3,3’-diol in ameliorating liver injury caused by indomethacin and by extension, may be adopted in the management of liver related injury.

Quantitative analysis of islet prohormone convertase 1/3 expression in human pancreas donors with diabetes.

Islet prohormone-processing enzymes convert peptide hormone precursors to mature hormones. Defective beta cell prohormone processing and the release of incompletely processed peptide hormones are observed prior to the onset of diabetes, yet molecular mechanisms underlying impaired prohormone processing during the development of diabetes remains largely unknown. Previous studies have shown that prohormone convertase 1/3 (PC1/3) protein and mRNA expression levels are reduced in whole islets from donors with type 1 diabetes, although whether PC1/3-mediated prohormone processing in alpha and beta cells is disrupted in type 1 diabetes remained to be explored. Herein, we aimed to analyse the expression of PC1/3 in islets from non-diabetic donors, autoantibody-positive donors and donors diagnosed with type 1 diabetes or type 2 diabetes. Immunostaining and high-dimensional image analysis were performed on pancreatic sections from a cross-sectional cohort of 54 donors obtained from the Network for Pancreatic Organ Donors with Diabetes (nPOD) repository, to evaluate PC1/3 expression patterns in islet alpha, beta and delta cells at different stages of diabetes. Alpha and beta cell morphology were altered in donors with type 1 diabetes, including decreased alpha and beta cell size. As expected, the insulin-positive and PC1/3-positive areas in the islets were both reduced, and this was accompanied by a reduced percentage of PC1/3-positive and insulin-positive/PC1/3-positive cells in islets. PC1/3 and insulin co-localisation was also reduced. The glucagon-positive area, as well as the percentage of glucagon-positive and glucagon-positive/PC1/3-positive cells in islets, was increased. PC1/3 and glucagon co-localisation was also increased in donors with type 1 diabetes. The somatostatin-positive cell area and somatostatin staining intensity were elevated in islets from donors with recent-onset type 1 diabetes. Our high-resolution histomorphological analysis of human pancreatic islets from donors with and without diabetes has uncovered details of the cellular origin of islet prohormone peptide processing defects. Reduced beta cell PC1/3 and increased alpha cell PC1/3 in islets from donors with type 1 diabetes pinpointed the functional deterioration of beta cells and the concomitant potential increase in PC1/3 usage for prohormone processing in alpha cells during the pathogenesis of type 1 diabetes. Our finding of PC1/3 loss in beta cells may inform the discovery of new prohormone biomarkers as indicators of beta cell dysfunction, and the finding of elevated PC1/3 expression in alpha cells may encourage the design of therapeutic targets via leveraging alpha cell adaptation in diabetes.

Curcumin Modulates the Differential Effects of Fructose and High-Fat Diet on Renal Damage, Inflammation, Fibrosis, and Lipid Metabolism.

Dyslipidemia and obesity hypercaloric diet-induced lead to kidney damage. We investigated the effect of curcumin on the expression of proteins related to inflammation, fibrosis, fatty acids metabolism, kidney damage, and morphological changes in the kidneys of mice hypercaloric diets-fed. Groups of 5-week-old C57BL/6 mice (n=6) were formed: Control (C), High-fructose diet (F), Highfructose diet and curcumin (F+Cur), High-fat diet (HFD), High-fat diet and curcumin (HFD+Cur), High-fat diet and fructose (HFD+F), High-fat diet, fructose and curcumin (HFD+F+Cur), treated for 16 weeks with 30% (w/v) fructose, 60% (w/w) fat and 0.75% (w/w) curcumin. Kidneys were obtained for histomorphological and Western Blot analysis. Curcumin prevented TNF-α overexpression in the F and HFD+F groups. VLCAD expression was higher in the F, HFD, and HFD+F groups. PPARγ expression was lower in the F+Cur, HFD+Cur, and HFD+F+Cur groups. Curcumin prevented overexpression of CPT1 and KIM1 in the HFD+F and HFD groups. Curcumin prevented morphological lesions, fibrosis, and lipid deposition that were hypercaloric diet-induced. Chronic consumption of hypercaloric diets causes inflammation, fibrosis, and lipid deposition in the kidney. It is suggested that curcumin prevents renal structural damage, limits tissue lipid deposition, and differentially modulates renal injury depending on diet composition in mice fed high-fat and/or high-fructose diets.

The therapeutic potential of reduced graphene oxide in attenuating cuprizone-induced demyelination in mice.

Reduced Graphene Oxide (rGO) has unique physicochemical properties that make it suitable for therapeutic applications in neurodegenerative scenarios. This study investigates the therapeutic potential of rGO in a cuprizone-induced demyelination model in mice through histomorphological techniques and analysis of biochemical parameters. We demonstrate that daily intraperitoneal administration of rGO (1 mg/ml) for 21 days tends to reduce demyelination in the Corpus callosum by decreasing glial cell recruitment during the repair mechanism. Additionally, rGO interferes with oxidative stress markers in the brain and liver indicating potential neuroprotective effects in the central nervous system (CNS). No significant damage to vital organs was observed, suggesting that multiple doses could be used safely. However, further long-term investigations are needed to understand rGO distribution, metabolism, routes of action and associated challenges in central neurodegenerative therapies. Overall, these findings contribute to the comprehension of rGO effects in vivo, paving the way for possible future clinical research.

Comparative histological and ultrastructural studies of thymus of indigenous and cross-bred broiler breeds of poultry of India

The present study was aimed to compare structure of thymus between the two poultry breeds, namely, Kadaknath and Narmada Nidhi. The aim was to understand the anatomical differences which could provide insights into how different breeds respond to diseases. The study involved histomorphological analysis of thymus tissue samples from 72 chicks (36 from each breed) that were divided into 6 groups with 6 birds in each group. The study results revealed that both poultry breeds exhibited similarities in the histology and ultrastructure of the thymus. This similarity is likely due to the Narmada Nidhi breed having a 25% genetic contribution from the Kadaknath breed.

Effects of Vitamin C on Aluminum Chloride- Induced Neurotoxicity on the Hippocampal Cortex of Adult Wistar Male Rats

This study aimed to evaluate the neuroprotective effects of Vitamin C on aluminum chloride (AlCl₃)- induced brain damage, focusing on behavioral, biochemical, and histomorphological outcomes in a rodent model. Study Design: A total of 42 healthy male rats were randomly assigned to three groups: control, AlCl₃ -only, and AlCl₃ plus Vitamin C (500 mg/kg and 1000 mg/kg). The AlCl₃ groups received daily doses of AlCl₃, while the Vitamin C groups received concurrent Vitamin C supplementation. Methodology: Body weight, behavioral changes, biochemical markers of oxidative stress (MDA, SOD, GSH), and inflammation (TNF-α, IL-6) were assessed. Histomorphological analysis of the hippocampus was performed to evaluate structural damage. Behavioral assessments included locomotor and exploratory activity tests. Biochemical analyses measured oxidative stress and antioxidant enzyme activities. Results: AlCl₃ administration resulted in significant body weight loss, increased oxidative stress (elevated MDA, and SOD levels), and heightened inflammation (elevated TNF-α and IL-6). Behavioral tests showed increased excitatory activities, and increased anxiety levels as seen in increased rearing and grooming activities. Histomorphological examination revealed significant hippocampal damages as seen in the pale staining, shrunken pyramidal cells. Vitamin C co- administration mitigated these adverse effects, demonstrating reduced body weight loss, lower oxidative stress, decreased inflammation, and improved behavioral performance. Histological analysis indicated less hippocampal damage in the Vitamin C treated groups. Conclusion: As an antioxidant, Vitamin C effectively attenuates the neurotoxic effects of AlCl₃ by reducing oxidative stress and inflammation, and by protecting the hippocampal cellular integrity. These findings suggest that Vitamin C holds potential as a therapeutic agent for mitigating neurotoxicity induced by aluminum compounds.

Impacts of dietary Ajwain (Trachyspermum ammi L.) on growth, antioxidative capacity, immune responses, and intestinal histology of grey mullet (Liza ramada)

This study investigated the impacts of dietary supplements with ajwain seed extract (ASE) on the growth, biochemical parameters, digestive enzymes, antioxidant activity, immunity, and intestine histology of Liza ramada. Juveniles (N = 450) with an initial weight of 40.12 ± 0.14 g were fed six experimental diets enriched with ASE at 0 (control), 0.2, 0.4, 0.6, 0.8, and 1.0 g/kg diet for 60 days. Results revealed that fish-fed diets containing ASE at levels of 0.6 g/kg or higher exhibited significantly improved growth parameters including final weight, weight gain percentage, and specific growth rate, along with enhanced feed conversion efficiency compared to lower ASE levels and the control diet. Survival rates remained similar across all experimental groups. Digestive enzyme activity, particularly lipase, was highest in groups fed higher ASE levels (≥ 0.4 g/kg diet). Histomorphological analysis of the intestine showed improved villi structures with increasing ASE supplementation, reaching an optimal appearance at 0.8 g/kg. Blood biochemical analysis indicated higher total protein and globulin concentrations, and lower total cholesterol levels in groups with higher ASE intake (≥ 0.4 g/kg diet). Immune parameters increased with increasing ASE concentrations, including lysozyme activity, bactericidal percentage, and nitroblue tetrazolium levels. Antioxidant status, measured through superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels, showed significant improvements with ASE supplementation, with optimal dietary ASE levels for maximizing SOD (0.73 g/kg), CAT (0.74 g/kg), GPx (0.83 g/kg), and minimizing MDA (0.73 g/kg) identified through polynomial regression analysis. These findings suggest that ASE supplementation (0.6–0.8 g/kg) positively influences the growth performance, digestive function, gut health, immunity, and antioxidant status of L. ramada, highlighting its potential as a beneficial dietary additive in aquaculture.

Arterial Histology in Amputated Limbs with Chronic Limb-Threatening Ischemia and Cadaver Limbs

ObjectiveThe study objectives were to 1) elucidate the histopathological and structural alterations in the femoral to popliteal arteries and tibial or more distal arteries in patients with chronic limb-threatening ischemia through detailed histomorphological and histopathological examinations of the lower limb arteries in the amputated limbs; and 2) elucidate the basic structure of the entire lower extremity, from the external iliac artery to the digital artery, based on histomorphological examinations of cadaver limbs. MethodsWe examined the histopathology and histomorphology of lower limb arteries harvested from 22 amputated limbs and that from 11 cadavers without lower limb issues. Sixty-three sections from the amputated limbs were categorized into femoropopliteal, tibial, and distal segments. Five hundred ten sections (from the external iliac artery to the digital artery) from the cadavers were categorized as femoropopliteal segments, tibial and distal segments, and others. The modified American Heart Association classification was used for the histopathological analysis, and the advanced lesion was defined as the presence of progressive atherosclerotic lesion. In the histomorphological analyses, we calculated %stenosis, %medial area as the medial area divided by the area within the external elastic lamina × 100, and %intimal area as the intimal area divided by the area within the external elastic lamina × 100. ResultsIn amputated limbs, advanced lesions and calcification in the medial area were more prevalent in the femoropopliteal segments than in the tibial and distal segments (advanced lesion: 8/10 vs. 18/42, p=0.048; calcification: 10/10 vs. 28/42, p=0.037), while %stenosis and prevalence of luminal thrombi were comparable. Significant inverse correlations were noted between %medial area and external elastic lamina diameter in both amputated (r=-0.641, p<0.001) and cadaver limbs (r=-0.775, p<0.001). In the femoropopliteal segments of the amputated limbs, luminal stenosis was solely caused by intimal lesions, whereas in the tibial and distal segments, medial area thickening contributed to luminal stenosis when intimal lesions were less pronounced. When the external elastic lamina diameter was entered as a covariate, the medial area of the 178 cadaver tibial and distal segments with minimal intimal lesions (<40% intimal area) was significantly greater when >75% stenosis was present in the femoropopliteal segments (p<0.001). ConclusionAdvanced lesions in the tibial and distal segments are less prevalent compared to the femoropopliteal segments. Medial area thickening may contribute to luminal stenosis in the tibial and distal segments when intimal lesions were less pronounced, which may be secondary to a significant proximal occlusive lesion.

Demonstrating the principal mechanism of action of medical devices intended for vaginal use on reconstructed human vaginal epithelium: the case of two hyaluronic acid-containing devices

Regulation (EU) 2017/745 on medical devices (MDR) has significantly modified the rules to be adopted for MD qualifications and classification. New requirements require robust evidence on mechanisms of action (MoAs) that cannot be produced by existing common EU or ISO standards. Therefore, on a “case-by-case basis,” a new evidence-based non-clinical approach to MD qualification must be defined. In this study, an in vitro experimental approach is described to assess the physicochemical and mechanical MoA of two hyaluronic acid (HA)-based medical devices: Mucogyne® Gel and Mucogyne® Ovule for vaginal use. They both act as moisturizers and lubricants as well as a healing adjuvant by promoting the continued moisture of the vulvovaginal area. The MoA of these two products has been demonstrated by using a 3D reconstructed human vaginal epithelium (HVE) model in a homeostatic physiological state and in stressed conditions. Film forming and persistency properties were assessed on intact HVE tissues by caffeine permeation assay and Lucifer Yellow (LY) localization on HVE vertical sections. Healing properties were assessed on injured HVE tissues by trans-electrical epithelial resistance (TEER) measurements associated with histo-morphological analysis (H&amp;amp;E), and moisturizing efficacy was evaluated on HVE tissues cultured in dry conditions by histomorphological analysis (H&amp;amp;E) and aquaporin 3 (AQP3) expression and localization by immunohistochemistry (IHC). Using the same “dry” HVE model, the non-pharmacological action of the two products was addressed by CD44 (hyaluronic acid receptor) expression and localization. The results suggest that in vitro evaluations can provide robust results on a human-relevant experimental model for the intended use of the products and supports clinical data with mechanistic information which may not be achieved with in vivo studies but are particularly important for product qualification. The results also underline the specific relative efficacy of the mechanisms investigated for Mucogyne® Gel and Mucogyne® Ovule in line with their different formulation types (respectively, hydrophilic and lipophilic) that influence the action of the active ingredient HA. The present in vitro non-clinical evaluation of HVE combined with clinical investigation data obtained in women explain why Mucogyne MDs provide significant benefits in various physiological or pathological situations, including vaginal dryness and healing.

Fgf9 promotes incisor dental epithelial stem cell survival and enamel formation

BackgroundUnderstanding the role of cytokines in tooth development is critical for advancing dental tissue engineering. Fibroblast growth factor 9 (FGF9) is the only FGF consistently expressed throughout dental epithelial tissue, from the initiation of tooth bud formation to tooth maturation. However, mice lacking Fgf9 (Fgf9−/−) surprisingly show no obvious abnormalities in tooth development, suggesting potential compensation by other FGFs. Here we report findings from an Fgf9S99N mutation mouse model, a loss-of-function mutation with a dominant negative effect. Our study reveals that Fgf9 is crucial for dental epithelial stem cell (DESC) survival and enamel formation.MethodsTo dissect the role of Fgf9 in tooth development, we performed the micro-CT, histomorphological analysis and gene expression assay in mice and embryos with S99N mutation. In addition, we assessed the effect of FGF9 on the DESC survival and dental epithelial differentiation by DESC sphere formation assay and tooth explant culture. Cell/tissue culture methods, gene expression analysis, specific inhibitors, and antibody blockage analysis were employed to explore how Fgf9 regulates enamel differentiation and DESC survival through both direct and indirect mechanisms.ResultsThe Fgf9S99N mutation in mice led to reduced ameloblasts, impaired enamel formation, and increased apoptosis in the cervical loop (CL). DESC sphere culture experiments revealed that FGF9 facilitated DESC survival via activating ERK/CREB signaling, without affecting cell proliferation. Furthermore, in vitro tissue culture experiments demonstrated that FGF9 promoted enamel formation in a manner dependent on the presence of mesenchyme. Interestingly, FGF9 stimulation inhibited enamel formation in isolated enamel epithelia and DESC spheres. Further investigation revealed that FGF9 supports DESC survival and promotes amelogenesis by stimulating the secretion of FGF3 and FGF10 in dental mesenchymal cells via the MAPK/ERK signaling pathway.ConclusionsOur study demonstrates that Fgf9 is essential for DESC survival and enamel formation. Fgf9 performs as a dual-directional regulator of the dental enamel epithelium, not only inhibiting DESC differentiation into ameloblasts to preserve the stemness of DESC, but also promoting ameloblast differentiation through epithelial-mesenchymal interactions.

Developmental variations of the reproductive organs of ganders from different goose breeds and the underlying mechanisms

A deep understanding of the dynamics and mechanisms of male reproductive tract development is necessary for adoption of either genetic techniques or environmental management practices for improving fertility and hatchability in poultry. However, compared with other poultry such as chickens and ducks, less is known about the age- and breed-related changes in the reproductive tract development of domestic goose ganders exhibiting relatively poor reproductive performance as well as the regulatory mechanisms. In the present study, by taking 2 Chinese domestic goose breeds (Sichuan White goose, SW and Gang goose, GE; Anser cygnoides) and one European goose breed (Landes goose, LD; Anser anser) as the experimental objects, we comprehensive analyzed the morphological, histological, and genome-wide transcriptomic variations in their testicular and external genital development during the period from hatching to sexual maturity. Results from histomorphological analysis demonstrated that the reproductive tract of all goose breeds developed in both age- and breed-dependent manners, and the left and right testis developed asymmetrically throughout posthatch development. The tenth week posthatch was a critical developmental stage for all goose ganders, because both the testicular and external genital histomorphological parameters significantly changed before and after this period. During the first 10 wk posthatch, the weight, organ index, or size of male reproductive organs developed more rapidly in SW than in LD, and so were the testicular parenchymal-to-interstitial ratio and the external genital lymphatic lumen diameter. However, the testicular seminiferous epithelium thickness, seminiferous tubule diameter, and Leydig cell number, as well as the external genital keratinized epithelium thickness were significantly higher in LD than in SW at 10 wk of age. Through comparative transcriptomics analysis and RT-qPCR validation, several pathways related to germ and somatic cell function, organ remodeling, and energy metabolism were thought to be responsible for the developmental variations in the early testicular development between Chinese and European domestic ganders, where 10 hub genes involved in the cell cycle, RNA polymerase II-dependent transcription, and mitotic cell division pathways might play essential roles. These data shed new light on the interbreed differences in the male goose reproductive tract development and the molecular mechanisms regulating male goose testicular functions and fertility.

A histomorphological and immunohistochemical analysis of invasive lobular carcinoma of breast: A case series

Invasive lobular carcinoma (ILC) is the second most common special subtype of invasive breast carcinoma comprising 5–15%. ILC defines as invasive carcinoma comprising non-cohesive single-file linear pattern in fibrous stroma. The aim of the study was to evaluate the histomorphological features and immunohistochemical analysis of ILC. A total of 19 cases were encountered from March 2020 to February 2024 out of 129 breast cancers. The sociodemographic profiles and clinicopathological parameters of the patients were studied with special emphasis on their morphology and hormone receptor status. The mean age of patients is 42.8 years. Out of 19 cases, 84.2% showed feature of classic type, 5.2% of tubule-lobular, and 10.5% of pleomorphic lobular carcinoma. The majority of cases (63.2%) showed histologic Grade 2, and 10.5% showed Grade 3. On immunohistochemistry examination, 79% were estrogen receptor (ER) and progesterone receptor (PR) positive with 10.5% of cases was human epidermal growth factor receptor 2/neu (HER2/neu) positive. Our study showed higher cases of classic type ILC, mostly of histologic Grade 2; higher ER-PR receptor positivity and HER2/neu negativity.

Hepatocyte-Specific Yap1 Knockout Maintained the Liver Homeostasis of Lipid Metabolism in Mice.

Yes-associated protein 1 (YAP1) is a crucial molecule in the Hippo pathway. The impact of hepatocyte-specific Yap1 knockout (Yap1 LKO) on hepatic lipid droplets (LD) and pePLIN2 in metabolic fatty liver has not been reported. This study aims to explore whether Yap1 LKO could offer a protective effect in a liver injury model. Three-week-old Yap1 LKO and Yap1 Flox mice were given aristolochic acid I (AAI) combined carbon tetrachloride (CCL4) establish liver injury model. Eight-week-old Yap1 LKO and Yap1 Flox mice were fed with a high-fat diet for 18 weeks to establish obesity-related liver injury model. Further biochemical, histomorphological, immunohistochemical, and lipidomic analyses were performed on serum and liver tissues of these mice to elucidate the effects of hepatocyte-specific Yap1 knockout on hepatic lipid metabolism. Yap1 LKO reduced triglyceride (TG) content and PLIN2 expression level in the liver during the intervention of AAI combined CCl4. Moreover, Yap1 LKO improved lipid metabolism homeostasis in the liver by increasing the beneficial lipid molecules and reducing the harmful lipid molecules through lipidomics. Finally, Yap1 LKO reduced TG content in the serum and liver, hepatic vacuolar degeneration, and hepatic PLIN2 expression level in mice fed with a high-fat diet (HFD). Yap1 LKO is protective in regulating liver and blood TG when induced with toxic substances AAI combined CCl4 and a high-fat diet.

Enhancing Clinicopathological Diagnosis of Hydatidiform Mole Through the Combined Application of Histomorphologic Analysis, Immunohistochemical Analysis of p57 Expression, and Short Tandem Repeat Typing Method

This study aims to assess the early histomorphologic characteristics and investigate the role of the p57kip2 protein combined with STR genotyping for pathological diagnosis and typing of the hydatidiform mole (HM). A total of 73 induced abortion tissues were collected for pathological evaluation, including 14 cases with partial HM (PHM), 7 cases with complete HM (CHM), and 52 cases with non-molar pregnancies. Histopathological examination of moles was conducted using hematoxylin and eosin staining. DNA extraction from paraffin sections was performed using Fe3O4 nano-magnetic beads. Molecular diagnosis was performed using STR genotyping. Immunohistochemical analysis was used to determine the distribution and expression level of p57kip2 protein in HM. Significant differences were observed in the morphological indices of villous edema, cistern formation, trophoblastic inclusions, and trophoblastic hyperplasia between the PHM and CHM groups (P &lt; 0.05). The central cistern formation and the trophoblast inclusion showed a significant difference between the HM and non-molar pregnancy (P &lt; 0.05). Moreover, our findings revealed that p57kip2 expression contributed to distinguishing CHM from PHM. However, it could not distinguish PHM from non-mole pregnancy. Furthermore, the results of STR genotyping were consistent with pathological typing. In conclusion, the integration of pathomorphology, immunohistochemical staining, and molecular diagnostics holds great value for the diagnosis and classification of HM.