Histological, histochemical, and immunohistochemical characterization of the gill skeleton in the silver carp (Hypophthalmichthys molitrix).

Exploring potential mechanism of Chuanzhi Qingyu recipe for vulnerable atherosclerosis plaques based on UPLC-Q-TOF-MSE, systems biology strategy, and experimental validation.

Objective To investigate the role and potential mechanism of CD200 in the pathogenesis of preeclampsia (PE) and to clarify its potential value as a biomarker candidate and possible therapeutic target for PE. Methods This retrospective study enrolled 57 patients with PE and 49 women with normal full-term pregnancies. Clinical data were collected, and serum biomarkers were measured. The GSE93839 dataset was utilized to screen for differentially expressed genes in PE placental tissues. Hematoxylin-eosin (HE) staining, immunohistochemistry (IHC), and immunofluorescence (IF) techniques were employed to analyze the expression and localization characteristics of CD200 in placental tissues. Exploratory path analysis was conducted to investigate potential indirect associations between CD200 expression and PE through serum biomarkers. Results Bioinformatics analysis identified CD200 as a candidate downregulated gene associated with PE. Immunohistochemistry revealed a significantly lower expression level of CD200 in the placental tissues of PE patients (p < 0.05). Immunofluorescence experiments demonstrated co-localization of CD200 with CD68 + macrophages, suggesting a potential association with macrophage-related immune regulation at the maternal-fetal interface. Exploratory path analysis revealed statistically significant indirect associations between CD200 expression and PE through serum biomarkers, including high-density lipoprotein cholesterol (HDL-C), albumin, alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB (CK-MB) (p < 0.05). Conclusion The expression of CD200 is significantly downregulated in the placental tissues of PE patients. This alteration may induce immune imbalance at the maternal-fetal interface and placental dysfunction, and may also be linked to the systemic pathological injury process in PE through its associations with multiple serum biomarkers.

Benign prostatic hyperplasia (BPH) is a widespread urological clinical case in aged men associated with troublesome lower urinary tract symptoms and cardiac dysfunction, but its medical treatment is still insufficient. Thus, searching for more effective medical regimens is an essential target. We tried in current experiment to explore the role of bosentan (BOS) and/or finerenone (FIN) in this lesion. BPH was adequately induced using testosterone propionate (TP) in a dose of (5mg/kg/day) for 28 days of subcutaneous injection and an oral administration of BOS (50mg/kg/day) and/or FIN (10mg/kg/day) for 10 days. We found typical histopathological changes of BPH, increased serum dihydrotestosterone (DHT), prostatic specific antigen (PSA), cardiac enzymes, tissue malondialdehyde (MDA), aldosterone, endothelin-A, nuclear factor kappa b (NF-κB), and proliferating cell nuclear antigen (PCNA). However, significant decreases in total antioxidant capacity (TAC), reduced glutathione (GSH), and caspase-3. The histological findings were augmented by special staining techniques, immunohistochemical and morphometric studies. There are histopathological alternations as the hyperplastic nodules in the prostate's transition zone expand, and the prostatic urethra was compressed, lengthened and distorted. Interestingly the co-administration of BOS and/or FIN significantly improved BPH induced changes. This is mostly attributed to its ability in modulating endothelin receptors by BOS, aldosterone receptors by FIN, improved the histopathological changes with anti-oxidant, anti-inflammatory, anti-fibrotic, antiproliferative, and apoptotic properties. BOS and/or FIN showed a significant ameliorative effect in BPH.

ASO Author Reflections: Expanding the Boundaries of ICG-Guided Anatomical Liver Resection: The "Jigsaw-Pattern" Positive Staining Technique.

In sexual assault investigations, the detection of spermatozoa from victims' clothing or vaginal samples constitutes critical evidence indicative of ejaculation. Staining methods such as Baecchi's staining and fluorescence immunostaining are widely used to enhance microscopic examination for sperm detection. However, these approaches have inherent limitations, including limited specificity (e.g., Baecchi's staining) and reduced performance in aged samples (e.g., immunostaining), underscoring the need for novel staining strategies. In the present study, we developed a dual-staining method targeting the protamine-DNA complex in sperm heads, utilizing a thiol-reactive fluorescent dye in combination with a DNA-binding fluorescent dye. This CPM/AO dual staining method produced strong green fluorescence localized to sperm heads under ultraviolet excitation. Based on the finding that sperm heads exhibited stronger fluorescence than somatic cell nuclei, this phenomenon may involve Förster resonance energy transfer (FRET). Although the nuclei of oral and vaginal epithelial cells, as well as other debris, exhibited similar coloration of the sperm head, spermatozoa could be effectively distinguished based on fluorescence intensity and related features, enabling reliable visual identification. Moreover, the method demonstrated consistent effectiveness in detecting spermatozoa in aged forensic specimens. These findings indicate that the proposed staining technique serves as a useful tool for sperm detection in forensic contexts, particularly when dealing with degraded samples.

To explore effects of needle knife on autophagy and matrix degradation-related proteins of chondrocytes in rabbits with knee osteoarthritis (KOA). Eighteen healthy male New Zealand rabbits aged 6 months, with a body weight of (2.0±0.5) kg, were selected and randomly divided into blank group, model group and needle knife group, with 6 rabbits in each group. KOA rabbit animal model was established by fixing and immobilizing left hind limb of rabbits in an extended position with a plaster cast for 6 weeks, following the modified Videman method. The control group was not immobilized. Both of control group and model group were not intervened. The needle knife group received needle knife intervention once a week for 4 weeks. One week later, Kellgren-Lawrence(K-L) grading and Magnetic Resonance Knee Osteoarthritis Score (MOAKS) were used to evaluate bone spurs, joint space, cartilage smoothness and effusion on the left knee joint of rabbits; Lequesne MG score was used to evaluate effects of acupuncture knife on symptoms, signs and functional activities of rabbit's knee joint. After killing rabbits, cartilage morphology of left knee joint was observed;pathological changes of cartilage on the left knee joint of rabbits were examined by hematoxylin O-eosin staining under a light microscope;TdT-mediated dUTP nick end labeling (TUNEL) staining technique was used to detect apoptosis rate of chondrocytes on the left knee joint of rabbits. Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot were used to detect autophagy of chondrocytes on the left knee joint and expressions of regulatory molecules for cartilage matrix degradation, including myosin-like bcl2 interacting protein (Beclin-1), light chain 3 (LC3), miR-107, cysteine aspartate-specific protease 1 (Caspase-1), interleukin-1β (IL-1β), collagen typeⅡ(CollagenⅡ), matrix metalloproteinase-3 (MMP-3), and matrix metalloproteinase-13 (MMP-13), at mRNA and protein levels. Compared with blank group, the surface of left knee joint cartilage in model group showed defects, with unclear structure, reduced matrix staining, disordered cell arrangement, mostly in clustered distribution, significantly reduced in quantity, and chaotic and unclear marginal lines;the apoptosis rate of chondrocytes increased, while K-L grading level, MOAKS score, Lequesne MG score rose, and visual assessment score also increased. The expressions of Caspase-1, IL-1β, MMP-3, and MMP-13 mRNA and proteins in cartilage tissue were upregulated, and all the differences were statistically significant (P<0.01). The expressions of Beclin-1, LC3, miR-107, CollagenⅡmRNA, as well as Beclin-1, LC3Ⅱ/Ⅰ, and CollagenⅡproteins in cartilage tissue were all decreased, and the differences were statistically significant (P<0.01). Compared with model group, the surface of left knee joint cartilage in needle knife group was smoother, with fewer defects, clearer structure, more uniform and normal matrix staining, more orderly cell arrangement and an increase in cell number, and the ridges were clearer and more complete;K-L grade, MOAKS, and Lequesne MG scores decreased, the visual assessment score decreased(P<0.05), apoptosis rate of chondrocytes decreased, and mRNA and protein expressions of Caspase-1, IL-1β, MMP-3, and MMP-13 in cartilage tissue were all downregulated;all the differences were statistically significant (P<0.01);expressions of Beclin-1, LC3, miR-107, CollagenⅡmRNA, and Beclin-1, CollagenⅡproteins in cartilage tissue were significantly increased (P<0.01), and expression of LC3Ⅱ/Ⅰprotein in cartilage tissue was also increased, with a statistically significant difference (P<0.05). Based on the theory of meridians, acupuncture and knife therapy could promote chondrocyte autophagy by regulating the expression of proteins such as Beclin-1, LC3, miR-107, and Caspase-1, reduce apoptosis, inhibit the degradation of cartilage matrix, lower the level of inflammation, delay degeneration of articular cartilage, and exert functions of protecting cartilage and improving knee joint pain and functional disorders.

Sperm cryopreservation induces intracellular oxidative stress. Sesamol (SE), as an antioxidant, mitigates cryodamage caused by spermatozoa cryopreservation. This study was designed to evaluate the protective effects of SE-loaded nanostructured lipid carriers (SE-loaded NLCs) on frozen sperm quality. SE-loaded NLCs were synthesized using hot homogenization, followed by characterization for particle size, morphology, and encapsulation efficiency (EE). Normozoospermic semen samples were collected from 20 men and prepared according to the WHO 2021 guidelines. The samples were allocated into experimental groups of fresh, frozen-thawed control, SE-treated, blank NLCs, and SE-loaded NLCs. Following semen cryopreservation and thawing protocols, sperm quality was evaluated regarding motility, vitality, morphology, membrane and acrosome integrity, mitochondrial membrane potential (MMP), and DNA fragmentation using established staining and microscopy techniques. Statistical analysis was also performed at the significance level of 0.05. NLCs were successfully prepared with a mean particle size of 154 nm, zeta potential of -17.6 mV, and EE of 72 ± 2.64%. Fluorescence microscopy confirmed the efficient uptake of NLCs by spermatozoa within 60 minutes. The optimal SE dose was determined as 50 µM for cryoprotection. Sperm treated with SE-loaded NLCs showed improvements (p < 0.05) in motility, viability, membrane and acrosome integrity, MMP, and reduced (p < 0.05) DNA fragmentation than other groups. Application of SE-loaded NLCs showed accelerated protective effects compared with free SE, highlighting the associated potential in the enhancement of sperm quality during cryopreservation. SE-loaded NLCs effectively enhanced sperm quality indices during cryopreservation. Thus, SE-loaded NLCs represent a promising strategy to mitigate cryoinjury and improve fertility preservation outcomes.

Cryptococcosis is an opportunistic infection widely associated with people living with HIV. Some even consider it one of the indicators of possible HIV infection during the peak of the HIV epidemic. The study aimed to investigate the incidence of cryptococcosis among HIV patients attending the Infectious Disease Hospital (IDH), Kano. In the study, KOH preparation, Gram staining technique, Culture method, Indian Ink staining techniques and urease test were employed for the isolation and identification of Cryptococcus species. In the results, apart from Cryptococcus species, Candida species and molds like Aspergillus and Penicilium species were also isolated. The female patients, as well as those within the age group of 30-39 years old, were more at risk of the infection. In conclusion, Cryptococcus species remain one of the opportunistic pathogens among HIV patients, with a higher risk among females.

ABSTRACTBackground and AimsIntestinal parasitic infections pose significant public health challenges in developing countries. They are major concerns in rehabilitation centers for individuals with mental disabilities. However, little attention has been given to preventing these infections among the mentally disabled groups. This study aimed to determine the prevalence of intestinal parasitic infections and associated factors in psychiatric patients.MethodsWe conducted a cross‐sectional study at Debre Tabor Comprehensive Specialized Hospital, Northwest Ethiopia between August 2024 and October 2024, on 402 participants enrolled by convenient sampling technique. The sociodemographic and clinical data were collected by psychiatry specialists. Stool samples were examined using wet mount, formol‐ether concentration, and modified acid‐fast staining techniques. Data were analyzed using the SPSS software version 23. Bivariate and multivariate logistic regressions determined the association between variables. Factors with p < 0.05 were considered statistically significant.ResultsOverall, 26.6% (107/402); [95% CI: 22.1%–31.3%] of the study participants had intestinal parasitic infections. Ascaris lumbricoides,9% (36/402); [95% CI: 6.5%–12.2%], was the most prevalent. Rural residence AOR = 3.5 (95% CI: 1.06–11.75), Large family size AOR = 2.9 (95%CI: 1.93, 5.46), Drinking surface water AOR = 2.6 (95%CI: 1.65, 5.00), in accessibility of latrine utilization AOR = 2.3 (95%CI: 1.34, 5.20), being bare footed AOR = 3.5 (95% CI: 1.23, 9.96), consuming raw vegetables/fruits AOR = 6.8 (95% CI: 4.38,10.63), and untrimmed finger nails AOR = 2.5 (95% CI: 1.80, 6.56) were significant factors associated with intestinal parasitic infections in psychiatric patients.ConclusionThe study revealed the significant prevalence of intestinal parasitic infections in psychiatric patients. Rural residence, larger family size, drinking surface water, unavailability of latrine, not wearing shoes, consuming uncooked vegetables/fruits, and untrimmed fingernails are the significant risk factors favouring the acquisition and spread of intestinal parasitic infections in psychiatric patients.

With the expanding biomedical applications of metal nanoparticles (MNPs), the accurate and intuitive detection of their in vivo behavior in complex biological systems has become an urgent research priority. Silver staining, a label-free and highly sensitive histochemical technology, addresses this challenge by amplifying ultrasmall MNPs to a size detectable by optical or electron microscopy. This allows reliable detection of their microstructural distribution and furnishes essential technical support for MNP-related biomedical research. In this study, we review the research progress of silver staining technology and its applications in investigating in vivo behaviors. We first summarize the development history of silver staining technology, with a focus on silver staining protocols for ultrasmall MNPs (< 2nm). We then systematically discuss the application of silver staining either alone or combined with other staining techniques for bright-field and electron microscopy for visualizing the distribution of MNPs at the sub-organ, cellular, and organellar levels. Finally, we comprehensively analyze the technical merits and intrinsic limitations of silver staining, and future directions. This review clarifies the critical value of silver staining for elucidating the in vivo behaviors of nanoparticles, and aims to provide technical support for the high-quality advancement of MNPs in nanomedicine.

Dysregulation of podocyte autophagy, triggering serious impairment of podocyte integrity, is a possible determinant of IgAN. To decipher the significant roles of Beclin-1 and LC3B in IgAN in regulating cellular autophagy, we investigated the role of PI3K/AKT signaling in regulating podocyte autophagy of IgAN. To carry out this study, we obtained tissue samples from 80 patients with primary IgAN (IgAN group) and 20 patients with kidney stones (control group). The IgAN group was further classified according to the Haas pathological grading system. The 100 paraffin-embedded tissue samples underwent hematoxylin-eosin staining, immunofluorescence, special staining techniques, immunohistochemical analysis (AKT, FoxO1, Beclin-1, LC3B, nephrin and podocin), and transmission electron microscopy. The number of autophagosomes was generally lower in the IgAN group than in the control group, however, in the IgAN group, the number of autophagosomes showed an increasing trend, with higher Haas grade. Beclin-1 and LC3B expression levels were highly correlated, and they were positively correlated with Haas grading, with LC3B showing a "U-shaped" trend: initial low expression in early grades (I-II), progressive increase to peak at grade III, followed by a potential decline in advanced grades (IV-V). Meanwhile, AKT expression was markedly reduced in the early stages (grades I-II) but gradually increased in the late stages (grades III-V). The number of autophagosomes demonstrated a strong positive correlation with Beclin-1 and LC3B expression. Autophagy is a key player in the occurrence and progression of IgAN. Dysregulation of the autophagy pathway represents a core mechanism in the pathogenesis of IgAN, a finding supported by the strong positive correlation between Haas grading and the number of autophagosomes, as well as Beclin-1/LC3B expression.

The claspers are the copulatory organs in male elasmobranchs, responsible for directing the semen into the female cloaca (C). However, the microscopic morphology of the claspers remains poorly understood. This study describes the morphology of the claspers and clasper glands (CGs) of cururu stingray (Potamotrygon wallacei) at different maturational stages, including neonates, juveniles, and adult active males. These secondary sexual organs were analyzed anatomically and processed using standard histological and histochemical staining techniques. The claspers of P. wallacei have a tubular shape and a brown coloration with light spots, intensified in juveniles and active males. Claspers are flexible in neonates, show an increased rigidity in juveniles, and are fully rigid in adults. Gradual rigidity of the claspers results from mesenchyme (ME) replacement by cartilage followed by mineralization. The claspers are lined by an epidermis (EP) with mucous cells (MC) secreting neutral and acid carboxylate mucosubstances; spermatic groove (SG) with cells exhibiting neutral mucosubstances; vascularized connective tissue and musculature associated with nerve bundles. Such tissue characteristics can assist in penetration, protection, rotational movements, and opening of the claspers during copulation. The CG is recognizable in neonates and becomes progressively more structured through ontogeny, reaching full differentiation in adults. The CG is delimited by two external muscular tunics, with secretory units (SU) formed by a simple columnar epithelium, demonstrating a holocrine secretion with neutral and acid carboxylate mucosubstances, more intense in adults. This secretion can assist in sperm capacitation, nutrition, and motility. These results confirm that secondary sexual organs undergo morphophysiological changes during sexual maturation in P. wallacei.

To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) preconditioned with low-magnitude high-frequency (LMHF) vibration on temporomandibular joint osteoarthritis (TMJOA). BMSCs with/without LMHF vibration were intra-articularly injected into TMJOA model rats. After 8 weeks, joint diameter was measured. Pathological alterations were observed through histological staining techniques (CXCR4, SOX9). Invitro, BMSCs and chondrocytes were co-cultured with LMHF vibration. Chondrocyte proliferation was assessed by CCK-8. In a tumor necrosis factor-α (TNF-α)-induced inflammatory microenvironment, signal transduction-related and autophagy-related proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The injection of LMHF vibrated BMSCs increases the joint diameter and alleviates TMJ tissue damage. The OA model led to an elevation in CXCR4 levels and a reduction in SOX9 in condyle cartilage, while vibrated BMSCs upregulated both. Invitro experiments, appropriate cyclic sinusoidal dynamic mechanical vibration enhanced the proliferative capacity of chondrocytes co-cultured with BMSCs. In inflammation, the vibration group downregulated chondrocyte MMP-13 and upregulated chondrocyte Collagen II/BMP7/β-catenin/Atg3, as well as BMSC Collagen X/BMP7/β-catenin/CXCR4/Beclin-1/Atg5-all protein levels exceeded those in the LiCl group. For cartilage tissue engineering, LMHF vibration serves as a promising strategy to rapidly expand cells with a reinforced chondrogenic phenotype.

Anatomic liver resection traditionally relies on intraoperative ultrasound to identify hepatic veins as landmarks for parenchymal transection.1-3 Recently, portal territory staining with indocyanine green (ICG) has emerged as a key research focus in precision liver surgery, offering direct visualization of segmentation boundaries.4,5However, a standardized technique for staining complex, multi-segmental tumors is lacking. This study evaluated a novel multi-subsegmental positive staining technique to achieve precise resection in such cases. The study enrolled a 59-year-old man with a 7.5-cm hepatocellular carcinoma (HCC) occupying segments Ⅷ and Ⅳa. Preoperative three-dimensional reconstruction mandated resection of both segments for adequate margins. Informed consent was obtained from the patient and his family. Intraoperatively, under ultrasound guidance, the portal veins supplying segments Ⅷ and Ⅳa were selectively punctured. A 1:1000 diluted ICG solution was injected into each, generating a composite positive staining map to delineate the combined resection territory. This approach was termed the "jigsaw-patterned staining technique." The total operative time was 240 min, with the staining procedure requiring 30 min. Intraoperative blood loss was 50 mL. The fluorescent boundaries closely correlated with the subsequent ischemic planes after targeted pedicle division. Histopathology confirmed HCC with microvascular invasion and satisfied margins. At this writing, the patient has recovered without complications and has maintained a disease-free survival for more than 20 months. The "jigsaw-patterned" multi-subsegmental portal vein ICG staining technique is technically feasible and safe. It enables precise anatomic resection for complex HCC spanning multiple segments, optimizing oncologic margin control while sparing functional parenchyma.

Chromovitrectomy has revolutionized vitreoretinal surgery by enabling the visualization of nearly transparent intraocular structures. This review evaluates the clinical utility of key vital dyes, such as brilliant blue G, trypan blue, indocyanine green, and triamcinolone acetonide, focusing on their physicochemical properties, staining affinities, and safety profiles. Recent advancements in dye formulation, including high-density and multi-component preparations, have improved tissue selectivity and surgical handling. These pharmacological innovations are now complemented by digital visualization technologies, such as 3D heads-up displays and customizable spectral filtering, which enhance contrast perception and ergonomics. We further discuss specialized surgical strategies for identifying occult retinal breaks and sequential staining techniques, while addressing critical concerns regarding phototoxicity and macular protection. As the field moves toward integrating intraoperative OCT and artificial intelligence, chromovitrectomy remains a dynamic cornerstone of posterior segment surgery. Understanding the synergy between novel biocompatible agents and digital platforms is essential for optimizing surgical precision and patient outcomes.

Abstract Background Indocyanine green (ICG) fluorescence imaging is increasingly incorporated into robotic liver resections (RLR), yet clinical practice regarding timing, dosage, and staining techniques is divergent. Aims This international expert survey aimed to characterize current practices for ICG in RLR. Methods Experts in RLR were invited to participate based on surgical volume (experience of 50 RLR and 30 annual RLR). A 74-item questionnaire was developed following a literature search and reviewed by a steering committee. The survey addressed indications, timing, dosage, imaging technology, benefits, limitations, training, and future directions of ICG use. Responses collected between September and October 2025 were analyzed. Results Seventy experts from 19 countries completed the survey, corresponding to an 88% response rate. Centers performed a median of 180 annual liver resections, including 55 RLR. Most experts used ICG (96%) during RLR. Anatomical demarcation (91%), tumor localization (60%), and biliary anatomy assessment (60%) were the most frequent indications. 60% of experts use preoperative ICG, while intraoperative ICG is mainly administered for demarcation (67%) and biliary tract visualization (40%). Considerable heterogeneity exists in dosage, timing, and staining techniques, particularly in cirrhotic livers and for tumor localization. Only half of the experts had standard operating procedures, whereas 64% expressed the need for a higher degree of standardization. Reported benefits of ICG use included improved anatomical orientation, margin assessment, lesion detection, and support during complex resections. Perceived limitations included background fluorescence, tissue penetration and variable staining in diseased parenchyma. 80% anticipated improved outcomes with combined ICG and three-dimensional image-guidance. Conclusion ICG fluorescence is widely used in RLR and is an important cornerstone for precision-guided robotic liver surgery. Standardized clinical practice guidelines, structured training, and technological improvements in imaging and navigation systems are claimed to optimize its clinical use.

Comparative evaluation of beetroot and Mangosteen Peel extracts as natural alternatives to malachite green in the Kato-Katz method.

As a vital barrier organ of the human body, skin wound repair faces clinical challenges such as slow healing, scar hyperplasia, and infection. Although the wet healing theory has confirmed that a moist environment can enhance epidermal cell migration and reduce infection rates, the action mechanism of composite functional dressings still lacks systematic analysis. This study focuses on hyaluronic acid (HA) polysiloxane gel, exploring its action pathway in full-thickness wound repair through a 'molecule-cell-tissue' three-level system. By using material-skin interaction data simulation,in vitrocell scratch assays, and SD rat full-thickness wound models, combined with HE staining, Masson staining, and RT-qPCR techniques, it was found that hyaluronic acid polysiloxane gel combines the anti-inflammatory regulation function of hyaluronic acid with the microenvironment maintenance function of polysiloxane gel. This significantly promotes fibroblast migration, enhances the expression of collagen-encoding genes Col 1a1 and Col 3a1, and accelerates wound healing. Experimental results from the rat full-thickness skin defect model showed that the wound healing rate in the hyaluronic acid polysiloxane gel treatment group exceeded 70% within 9 d, with smooth new epidermis, significantly reduced inflammatory cells, and a notably higher collagen density compared to the dry dressing group. This study innovatively constructs a framework for the action mechanism of composite dressings guided by the wet healing theory, confirming that hyaluronic acid polysiloxane gel accelerates wound repair through the synergistic effect of inflammatory regulation and microenvironment maintenance. It provides a theoretical basis for the research and development of the new generation of functional dressings and enriches the molecular mechanism connotation of the wet healing theory.

Exploration of the regulatory mechanism of the vasoactive intestinal peptide-eosinophil axis in sheep resistance to Moniezia benedeni infection.