Histochemistry and Cell Biology最新文献

A growing body of evidence indicates that the endocannabinoid system (ECS) is essential for controlling the homeostasis of the skin and that the ECS is modified in the presence of skin disease. It is plausible to expect that the lamellar junction of the hoof expresses cannabinoid receptors and that their expression could be affected by lamellar disease. The goal of this study was to characterise the cannabinoid receptor type 1 (CB1R) and type 2 (CB2R) and the G protein-coupled receptor 55 (GPR55) within the dermo-epidermal junction of the hooves of healthy and laminitic horses. The expression of the CB1R, CB2R, and GPR55 within the dermo-epidermal lamellar junction of six healthy and 12 laminitic hooves was studied using polymerase chain reaction (PCR) and immunofluorescence. Both the mRNA and protein expression of the CB1R, CB2R, and GPR55 were found in the dermo-epidermal lamellar junction of horse hooves. The immunolabelling was expressed by the epithelial cells of the primary and secondary laminae of healthy hooves (CB2R > GPR55 > CB1R). The presence of CB1R, CB2R, and GPR55 immunoreactivity in the healthy laminar epithelial cells, coupled with increased protein expression in pathological epithelial cells, provided strong motivation for future investigation. These findings suggest that cannabinoid compounds which interact with these receptors may influence lamellar healing and mitigate inflammation in hoof diseases, particularly laminitis.

The success of both oogenesis and early embryo development relies heavily on dynamic epigenetic regulation in which gene activity changes without affecting the underlying DNA sequence. Epigenetics works through two main mechanisms: DNA methylation and histone modifications. DNA methylation typically leads to gene silencing, while histone modifications can either activate or repress genes depending on the specific modification, histone type, and targeted amino acid residue. Histone modifications affect important DNA regulatory processes in which the histone core area as well as the N-terminal tails that extend from the core region are vulnerable to a variety of posttranslational modifications (PTMs), including methylation, citrullination (deimination), acetylation, phosphorylation, ubiquitination, SUMOylation, ribosylation, and lactylation. This review article focuses on what is known about changes in the histone modifications and how these modifications and their responsible enzymes operate throughout mammalian oocyte maturation and early embryo development, highlighting their crucial roles in these processes.

The use of oxalic acid is common in metallic impregnation techniques, where it serves as a reducing and bleaching agent for potassium permanganate. Given that oxalic acid is harmful to health, its replacement with another reagent is advantageous for healthcare professionals handling it in laboratory settings. Considering that citric acid can also reduce potassium permanganate, the aim of this study was to determine whether oxalic acid in the Gomori reticulin technique could be replaced by citric acid, a compound substantially less hazardous to health. To this end, 1%, 5%, and 10% citric acid solutions were tested on porcine liver and kidney samples, and the intensity of reticulin fiber staining, contrast, and overall morphological preservation were assessed in comparison with 1% oxalic acid. The results showed no statistically significant differences between the positive control (oxalic acid) and the citric acid protocols, with the 5% citric acid concentration proving most favorable in terms of required incubation time and the evaluated morphological parameters. It was therefore concluded that citric acid can be used as a bleaching agent for potassium permanganate in the Gomori technique, effectively replacing oxalic acid.

Joint-related diseases often involve multiple aspects of angiogenesis, alterations in the extracellular matrix (ECM), and inflammatory responses, causing pain and mobility problems for patients. Angiopoietins (Angs) are a protein family of secreted growth factors that have a variety of functions, including regulating angiogenesis, stabilizing blood vessels, and mediating changes in the ECM. The first Ang was discovered by Davis in COS cells, and since then, three more members of this protein family have been discovered. Among these four members, Ang-1 and Ang-2 play the most essential roles in joint diseases and are closely related to each other as ligands for the Tie-2 receptor. Ang-1 acts as an agonist at the Tie-2 receptor, while Ang-2 can act as either an agonist or antagonist of Tie-2 under specific conditions. This review describes the structure and general physiological functions of Angs and summarizes the role of Angs in joint-related diseases. It concludes with possible Ang-based therapeutic strategies.

Super-resolution radial fluctuation (SRRF) microscopy is a novel computational imaging technique that bypasses the optical diffraction limit (lateral resolutions of 200-300 nm), achieving lateral resolutions of approximately 50-100 nm while being compatible with live-cell imaging. Unlike traditional super-resolution methods such as stimulated emission depletion (STED) and single molecule localization microscopy (SMLM), SRRF minimizes phototoxicity and hardware complexity by analyzing fluorescence intensity fluctuations in standard wide-field microscopy data. This is achieved by calculating local gradient convergence ("radiality") across time-series images, enabling the reconstruction of sub-diffraction structures without specialized fluorophores or high-intensity illumination. Implemented through the open-source NanoJ-SRRF platform, SRRF optimizes parameters like ring radius and radiality magnification to enhance resolution, suppress noise, and maintain computational efficiency. Its advantages include low phototoxicity, compatibility with conventional dyes, and integration with various imaging modalities, allowing dynamic visualization of subcellular processes (e.g., mitochondrial fission, microtubule dynamics). Despite its limitations in axial resolution and potential artifacts in high-density structures, recent advancements like enhanced SRRF (eSRRF) and variance reweighted radial fluctuations and enhanced SRRF (VeSRRF) address these challenges, facilitating real-time, multicolor imaging. Applications range from ultrastructural studies to clinical pathology, with future developments in AI processing and multimodal integration promising further enhancements in imaging capabilities. SRRF stands to significantly impact the understanding of dynamic subcellular processes and biomedical research.

Traumatic spinal cord injury (TSCI) is a severe event within the central nervous system, leading to the impairment of sensory and motor abilities at the injury site in the spinal cord. Current therapeutic approaches offer limited efficacy in improving recovery, underscoring the need to explore alternative treatments. In the present study, we examined the sustained co-delivery of basic fibroblast growth factor (bFGF) and interleukin-10 (IL-10), encapsulated in microspheres and incorporated into a collagen-based hydrogel, for its effects on the healing of TSCI in an animal model. Seventy-five male Sprague Dawley rats were randomly divided into five groups: control, TSCI, hydrogel, microspheres, and hydrogel loaded with microspheres (Hyd + Micro). Tissue samples were collected from the injury site for further evaluation. Compared to the TSCI group, treatment groups demonstrated notably higher numerical densities of neurons and greater motor neuron health, as well as increased levels of vascular endothelial growth factor (VEGF) and antioxidative factors (catalase [CAT], glutathione [GSH], and superoxide dismutase [SOD]), and improved neurological function scores (electromyography [EMG], Basso-Beattie-Bresnahan (BBB) test, and narrow beam-walking test [NBT]). These changes were most pronounced in the Hyd + Micro group. The treatment also led to a significant decrease in the counts of apoptotic and glial cells, alongside reduced levels of malondialdehyde (MDA) and pro-inflammatory cytokines (IL-1β and TNF-α), with the most notable enhancements seen in the Hyd + Micro group. These findings indicate that the co-delivery of bFGF and IL-10 encapsulated in microspheres within a collagen-based hydrogel provides enhanced neuroprotective effects in TSCI animal models.

MOTS-c is a promising regulator of metabolism and energy homeostasis. While its effects have been studied in cell lines, our team aimed to investigate its influence on more complex structures-specifically, isolated pancreatic islets. We used two animal models: the rat, which is commonly studied, and the pig, which shares greater physiological similarities with humans. This study assessed the expression and secretion of insulin and glucagon, the expression of their receptors, cell viability, and cell death following MOTS-c treatment of the islets. Additionally, we examined how MOTS-c secretion is affected by different incubation media, such as the presence of free fatty acids, pancreatic hormones, and different glucose concentrations. The results indicate that MOTS-c impacts pancreatic islet physiology by, for example, reducing insulin and glucagon secretion and enhancing cell viability. Notably, the effects differed between the two species, which may be attributed to anatomical differences in their pancreatic islets or structural variations in rat and pig MOTS-c. These facts may lead to the conclusion that if MOTS-c may be helpful in human medicine, the pig model should be considered another valuable choice.

In cancer tissue diagnostic studies of clinical medicine, cell morphology is a key indicator for assessing the behavior of cellular physiological activities. Currently, the method of using thermotherapy as an adjunct to cancer treatment has gradually become a trend. Assessing the morphological characteristics of wound tissue cells during the hyperthermia process is particularly important for providing feedback on the therapeutic efficacy of hyperthermia-assisted treatment. Among the existing cell observation techniques, optical bright-field microscopy can only perform static observations of cells from a two-dimensional planar perspective. However, fluorescence microscopy suffers from issues such as phototoxicity and low temporal resolution. To address the aforementioned issues, this study introduces a quantitative analysis method based on digital holography to overcome these limitations. According to the mechanism of hyperthermia in cancer treatment, cancer cells exhibit morphological changes when exposed to elevated temperatures. Digital holography technology can effectively utilize light refractive indexes and phase differences to quantify the thickness and volume of cells. This study systematically evaluated the morphological changes in HeLa cells and human cervical epithelial cells (HCECs) under different temperature gradients (37-42 ℃ and 60 ℃). Continuous tracking of cell thickness and volume was achieved. The results revealed a unique morphological thermoresponsive process of the cells. The study determined the temperature threshold and exposure duration for high-temperature-induced effects in HeLa cells. The greater temperature sensitivity of HeLa cells compared with HCECs has been verified. This technology is expected to provide an effective means for evaluating morphological changes in cellular thermoresponses, offering novel insights for optimizing personalized cancer treatment regimens.

Remodeling of the extracellular matrix (ECM) throughout the estrous cycle is one of the most striking features of the uterus. A disintegrin and metalloprotease with thrombospondin type I motifs (ADAMTS-1) is a metalloproteinase responsible for the degradation of some proteoglycans, which are ECM components. In this study, ADAMTS-1 distribution was analyzed in the uterus of ovariectomized rats administered 17β-estradiol (E2) and progesterone (P4) and in the uterus at different estrous stages. Ovariectomized (OVX) rats were subjected to single and combined E2 (0.2 mg/kg) and P4 (10 mg/kg) hormone replacement therapies. E2 was administered for 3 consecutive days, followed by E2, P4, or E2 + P4 for 4 consecutive days. The serum level of E2 decreased from the proestrus phase to the diestrus phase, but that of P4 was the highest in the estrus phase. During the estrus phase, the serum level of luteinizing hormone (LH) was the lowest and that of follicle-stimulating hormone (FSH) was the highest. P4 level increased significantly in the OVX + P4 and OVX + E2 + P4 groups compared with the OVX group. The serum levels of LH and FSH decreased in the OVX + E2 and OVX + P4 groups compared with the OVX group, and were the lowest in the OVX + E2 + P4 group. ADAMTS-1 immunoreactivity in luminal, glandular, and stromal cells of the uterus decreased from proestrus to diestrus. When immunoreactivity in hormone replacement groups was compared, weak immunoreactivity was observed in the OVX group. ADAMTS-1 immunoreactivity gradually increased in OVX + P4 and OVX + E2 groups, and was particularly notable in luminal, glandular, and stromal cells in the OVX + E2 + P4 group. ADAMTS-1 distribution was affected by the estrous cycle process and hormone replacement therapy in the OVX procedure.