Blue Staining Research Articles

Antibacterial and Antiallergic Effects of Three Tea Extracts on Histamine-Induced Dermatitis.

Atopic dermatitis (AD) is a persistent and recurrent inflammatory skin condition with a genetic basis. However, the fundamental reasons and mechanisms behind this phenomenon remain incompletely understood. While tea extracts are known to reduce histamine-induced skin allergies and inflammation, the specific mechanisms by which various types of Chinese tea provide their protective effects are still not fully elucidated. In this study, a model of skin itching induced by histamine is used to explore the functions and mechanisms of three types of tea extract (Keemun black tea (HC), Hangzhou green tea (LC), and Fujian white tea (BC)) in alleviating histamine-induced dermatitis. The components of three tea extracts are identified by UPLC-Q-TOF-MS, and we found that their main components are alkaloids, fatty acyls, flavonoids, organic acids, and phenols. The inhibitory effects of three types of tea extract on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) in skin injury are investigated by MIC and flow cytometry. The three types of tea extract have an inhibitory effect on the growth of bacterial flora, with HC showing the best inhibitory activity. The effect of the three types of tea extract on histamine-induced dermatitis is also evaluated. Furthermore, itchy skin experiments, HE staining, toluidine blue staining, and immunohistochemical staining of mouse skin tissues were performed to determine the variations of scratching, epidermal thickness, mast cell number, IL-1β, and NGF content after the administration of the tea extracts. The three types of tea extracts all alleviate and inhibit skin itching, epidermal hyperplasia, and allergic dermatitis. BC effectively alleviates epidermal hyperplasia caused by skin allergies, and LC significantly downregulates NGF. HC reduces histamine-induced mast cell infiltration and downregulates IL-1β to alleviate skin itching. Consequently, tea emerges a potent natural product that can inhibit the growth of skin wound bacterial flora and exhibit skin repair effects on histamine-induced allergic dermatitis.

Comparison of Neural Recovery Effects of Botulinum Toxin Based on Administration Timing in Sciatic Nerve-Injured Rats.

This study aimed to assess the effects of the timing of administering botulinum neurotoxin A (BoNT/A) on nerve regeneration in rats. Sixty 6-week-old rats with a sciatic nerve injury were randomly divided into four groups: the immediately treated (IT) group (BoNT/A injection administered immediately post-injury), the delay-treated (DT) group (BoNT/A injection administered one week post-injury), the control group (saline administered one week post-injury), and the sham group (only skin and muscle incisions made). Nerve regeneration was assessed 3, 6, and 9 weeks post-injury using various techniques. The levels of glial fibrillary acid protein (GFAP), astroglial calcium-binding protein S100β (S100β), growth-associated protein 43 (GAP43), neurofilament 200 (NF200), and brain-derived neurotrophic factor (BDNF) in the IT and DT groups were higher. ELISA revealed the highest levels of these proteins in the IT group, followed by the DT and control groups. Toluidine blue staining revealed that the average area and myelin thickness were higher in the IT group. Electrophysiological studies revealed that the CMAP in the IT group was significantly higher than that in the control group, with the DT group exhibiting significant differences starting from week 8. The findings of the sciatic functional index analysis mirrored these results. Thus, administering BoNT/A injections immediately after a nerve injury is most effective for neural recovery. However, injections administered one week post-injury also significantly enhanced recovery. BoNT/A should be administered promptly after nerve damage; however, its administration during the non-acute phase is also beneficial.

Level of Aflatoxins in Selected Cereals in Ekiti State, Nigeria

Aflatoxins are family of toxins produced by certain fungi found on agricultural crops such as maize (corn), peanuts, cottonseed, and nuts. The ingestion of such mycotoxin contaminated grains by animals and human beings has enormous public health significance. This has necessitated the evaluation of effects of Asparagus africanus (Lam - whole plant) extracts on aflatoxins production by fungi isolated from selected foods in Ekiti State, Nigeria. A total of 240 food samples were collected across the 16 Local Government areas in rural open markets. The food samples analyzed were: Igbemo rice Oryza sativa igbemo curtivar), maize (Zea mays) and groundnut (Arachis hypogaea). Aflatoxin-producing fungi and non aflatoxin producing fungi were isolated and identified using morphological method of identification on PDA and microscopic with lactophenol cotton blue stain. The molecular identification of the isolates was done using internal transcribe spacer (ITS) region. A total of 70 samples that contain Aspergillus spp were subjected to ELISA test to quantify and identify types of aflatoxins that were present in them. It was revealed that four types of aflatoxins were present in the food samples (AFB1, AFB2, AFG1, and AFG2). Also, different species of isolates were obtained from the samples viz: Aspergillus flavus, Aspergillus niger, Rhizopus stolonifer, Mucor mucedo, A. parasiticus, Neurospora crassa, Botrytis cinerea, Fusarium oxyporum and Penicillium oxalicum. Aspergillus flavus was prevalent among the isolates. The molecular identification showed three of the isolates were Aspergillus flavus with different percentage of ITS4 rDNA identity. The level of aflatoxins in some of the food samples analyzed is of great concern because of its ability to cause disease (in man and animal) and these toxins are introduced mostly during storage, that is, post-harvest.

Anti-Toxoplasma gondii effects of XYP1-derived peptides and regulatory mechanisms of XYP1

BackgroundToxoplasmosis, caused by Toxoplasma gondii , poses serious health issues for humans and animals. Individuals with impaired immune systems are more susceptible to severe toxoplasmosis. Pregnant women infected by T. gondii can face the possibility of birth defects and miscarriages. While pyrimethamine and sulfadiazine are commonly used drugs in clinical practice, concerns over their side effects and resistance are on the rise. A spider peptide XYP1 isolated from Lycosa coelestis had potent anti-T. gondii effects, but it had a high synthesis cost and strong cytotoxicity.MethodsThis study intended to modify XYP1 for producing derived peptides via amino acid truncation and substitution. The anti-T. gondii effect was evaluated by trypan blue staining assay and killing experiment of RH strain tachyzoites. The CCK8 and hemolysis assays were used to compare their safeties. The morphological changes of T. gondii were observed by scanning electron microscope and transmission electron microscope. In addition, the mechanism of XYP1 against T. gondii through RNA-sequencing was further explored.ResultsIn vivo and in vitro experiments revealed that XYP1-18 and XYP1-18-1 had excellent anti-T. gondii activity with lower cytotoxicity and hemolysis activity than XYP1. XYP1, XYP1-18, and XYP1-18-1 were able to disrupt the surface membrane integrity of T. gondii tachyzoites, forming pores and causing the disruption of organelles. Furthermore, RNA-sequencing analysis indicated that XYP1 could stimulate the host immune response to effectively eliminate T. gondii and lessen the host’s inflammatory reaction.ConclusionsXYP1-18 had lower cytotoxicity and hemolysis activity than XYP1, as well as significantly extending the survival time of the mice. XYP1 played a role in host inflammation and immune responses, revealing its potential mechanism. Our research provided valuable insights into the development and application of peptide-based drugs, offering novel strategies and directions for treating toxoplasmosis.Graphical

Killing of xenogenous and virally infected homogenous target cells by shrimp lymphocyte-like haemocytes

Haemocytes play a crucial role in the invertebrate's immune system. In our lab, five subpopulations of shrimp haemocytes were identified in the past: hyalinocytes, granulocytes, semi-granulocytes and two subpopulations of non-phagocytic cells. In the latter two subpopulations, their characteristics such as having small cytoplasmic rims and not adhering to plastic cell-culture plates are very similar to those of mammalian lymphocytes. Therefore, they were designated lymphocyte-like haemocytes. Although little is known about their function, we hypothesize, based on their morphology, that they may have a cytotoxic activity like natural killer cells, with the ability to recognize and kill target cells. In our study, K562 cells and Sf9 cells were used as xenogenous target cells to detect the cytotoxic activity of the shrimp non-adherent lymphocyte-like haemocytes. Non-adherent haemocytes were collected and mixed with K562 cells and Sf9 cells at a 5:1 ratio and the binding activity was examined under a microscope. The binding rate of non-adherent haemocytes to K562 cells and Sf9 cells reached 6.6 % and 2.4 % after 240 min of culture, respectively. Then, the killing activity of non-adherent haemocytes was detected by an EMA staining (fluorescence microscopy), which showed 3.75 % dead K562 cells and 1.025 % dead Sf9 cells, and by Sytox® blue staining (flow cytometry), which showed 4.97 % of dead K562 cells. Next, a killing assay was developed to visualize the killing activity of shrimp non-adherent haemocytes. Non-adherent haemocytes were pre-labeled in blue (CellTracker blue) and K562/Sf9 cells in green (CFSE); dead cells were differentially stained red with ethidium bromide. The cytotoxic activity increased and reached a level of 2.59 % in K562 cells and 0.925 % in Sf9 cells at 120 min after co-culture. Furthermore, in the co-cultures of non-adherent haemocytes with K562 cells and Sf9 cells, upregulation of the gene and protein expression of the cytotoxic molecules torso-like protein and granzyme B was observed by RT-qPCR at 240 min and western blotting at 180 min. Additionally, non-adherent haemocytes were co-cultured with WSSV-inoculated shrimp ovary and lymphoid organ cells to detect the cytotoxicity to homogenous target cells. The binding activity started at 60 min in both the ovary and lymphoid organ cultures and reached at 240 min 50.62 % and 40.7 %, respectively. The killing activity was detected by EMA staining and the percentage of dead ovary and lymphoid organ cells increased respectively from 10.84 % to 6.89 % at 0 min to 13.09 % and 8.37 % at 240 min. In conclusion, we demonstrated the existence of cytotoxic activity of shrimp lymphocyte-like haemocytes against xenogenous cells from mammals and insects and against WSSV-infected homogenous shrimp cells.

Studies on the Preparation of a Microemulsion Formulation of Matricaria Recutita Essential Oil and the Treatment of 2,4-Dinitro-Chlorobenzene- Induced Eczema in Mice by Inhibiting Inflammation.

Eczema, an inflammatory skin disease causing intense itching, is a function of a range of internal and external factors, impacting individuals of all ages and leading to economic loss. Inflammation is the most important manifestation of eczema, and Matricaria recutita essential oil (MREO) extracted from Matricaria recutita possesses excellent antibacterial and anti-inflammatory properties. In this study, Matricaria recutita microemulsions were prepared by the trans-phase emulsification method and their stability was determined by evaluating the relevant indexes. Establishment of 2,4-dinitro-chlorobenzene-induced AD model in mice. Detection of serum indexes of IL-6, IL-17, and TNF-α, and on pathological tissue sections, the HE staining, toluidine blue staining, immunohistochemistry, and observation were performed. The study obtained optimal conditions for the preparation of microemulsion formulations of Matricaria recutita. Through quality evaluation, it was found that the microemulsion increased stability, reduced irritation, and retained anti-inflammatory activity and therapeutic effects on eczema compared to Matricaria recutita essential oil (MREO). Studies have demonstrated that microemulsion formulations of Matricaria recutita and Matricaria recutita significantly down regulate the proinflammatory factors TNF-α, IL-17, and IL-6. It was shown by hematoxylin-eosin (HE) staining that both Matricaria recutita essential oil (MREO) and Matricaria recutita microemulsion (MRME) improved the inflammatory status of eczematous skin tissues in mice. The number of mast cells expressed in the tissues was decreased in the surface-treated group, as shown by toluidine blue staining. Additionally, the number of mast cells expressed in the tissues in the surface-treated group was reduced, as demonstrated by immunohistochemistry. Furthermore, immunohistochemistry revealed that MREO and MRME have immunomodulatory effects on the tissues. The study showed that microemulsion formulations of Matricaria recutita may serve as a novel remedy for eczema.

Rapid detection of guttae area using aniline blue staining in Fuchs endothelial corneal dystrophy mouse model.

Fuchs endothelial corneal dystrophy (FECD) is a leading cause of corneal endothelial degeneration resulting in impaired visual acuity. Excessive deposition of extracellular matrix (guttae) on Descemet's membrane (DM) is the hallmark of FECD. We sought to detect the guttae area rapidly using aniline blue (AB) staining in FECD mouse model. FECD mouse model was established via ultraviolet A (UVA) exposure. Masson's trichrome staining was utilized to stain the corneal sections. AB staining was utilized to stain both whole cornea tissues and stripped Descemet's membrane-endothelium complex (DMEC) flat mounts, while immunofluorescence staining of collagen I was employed to stain guttae areas. In Masson's trichrome staining, corneal collagen fibrils were stained blue with AB. The DMEC flat mounts were stained into relative dark blue areas and relative light blue areas using 2% AB staining. The areas of dark blue could almost overlap with collagen I-positive areas, and have an acellular centre and a moderately distinct boundary line with the surrounding corneal endothelial cells. In conclusion, AB staining is a rapid and effective method for the evaluation of the guttae areas in the FECD mouse model.

Effects of acupuncture on serotonin, histamine, substance P, and tryptase levels at sensitized points in model rats with knee osteoarthritis

ObjectiveTo elucidate the differences in manual acupuncture effectiveness at sensitized points by investigating the mechanisms of local skin action at different sensitization points in rats with knee osteoarthritis (KOA). MethodsForty Sprague–Dawley rats were equally divided into control, model (1 mg of monoiodoacetate into the right knee joint cavity), sham operation, manual acupuncture at right Tianjing acupoint (MAR-SJ 10), and left SJ 10 groups. Safranine-O and fast green staining were used to assess the modeling. The morphological and functional changes in mast cells (MCs) were assessed during acupoint sensitization using toluidine blue and immunofluorescence staining. The levels of serotonin, histamine, substance P (SP), and tryptase at skin acupoints and serum levels of IL-β, IL-6, and TNF-α were detected using ELISA. ResultsAfter 14 days of treatment, the number of MCs and their degranulation rates were statistically higher in the model group than in the control group (both P < .001). After applying acupuncture, the levels of 5-HT, HA, and SP at skin acupoints were lower than those in the model group (all P < .05), and tryptase level was higher (both P < .05). Tryptase level was higher on the skin at the MAL-SJ 10 acupoint than that on the MAR-SJ 10 acupoint (P = .004). Compared with the model group, the serum levels of IL-1β, IL-6, and TNF-α in the MAR-SJ 10 and MAL-SJ 10 groups were lower (all P < .05). ConclusionAcupuncture at KOA-sensitized acupoints mitigates joint injury in KOA rats and may bidirectionally regulate local MCs of these acupoints. This finding not only enhances the reference value of sensitizing points in clinical diagnosis and treatment, but also contributes to the understanding of the biological mechanisms underlying acupuncture intervention at sensitizing points.

Buyang Huanwu decoction promotes angiogenesis and improves hemorheological parameters after cervical spinal cord injury

ObjectiveTo explore the effects of Buyang Huanwu decoction (BYHWD) on vascular neogenesis and hemorheological parameters following cervical spinal cord injury (SCI). MethodsAn acute cervical SCI model was established using 84 female Sprague–Dawley rats. Functional recovery of the rats was evaluated using the forelimb locomotor scale score, forelimb grip strength test, and Basso-Beattie-Bresnahan score. The animals were subsequently euthanized at days 7 and 28 postoperatively. The gross morphology, neuronal survival, and myelin sheath in the injured area were evaluated using hematoxylin and eosin (HE), Nissl, and luxol fast blue (LFB) staining, respectively. Immunofluorescence staining was used to observe CD31 expression 7 days post-injury. Furthermore, the expression of CD31, neuronal nuclear protein (NeuN), and myelin basic protein (MBP) were evaluated 28 days post-injury. Additionally, vascular endothelial growth factor A (VEGFA) and VEGF receptor-2 (VEGFR-2) expression was evaluated using western blotting. Whole-blood viscosity, plasma viscosity, and red blood cell aggregation were measured using a hemorheometer. ResultsFrom postoperative days 3–28, motor function in the BYHWD group began to recover considerably compared to the SCI group. BYHWD effectively restored spinal cord histopathology. In addition, the number of NeuN-positive cells, and fluorescence intensity of CD31at 7 and 28 days and MBP significantly increased in the BYHWD group compared with the SCI group (all P < 0.05). Moreover, this decoction significantly upregulated the expression of VEGFA and VEGFR-2 (all P < 0.05). BYHWD improved the hemorheology results (i.e., except erythrocyte aggregation index in the low-dose group), revealing statistically significant differences compared with the SCI group (all P < 0.05). ConclusionBYHWD effectively promoted angiogenesis, improved hemorheological parameters, and protected neurons and myelin sheaths, ultimately promoting the recovery of neurological function after cervical SCI in rats. These findings suggest that BYHWD promotes vascular neogenesis through the VEGFA/VEGFR-2 pathway.

Embryonic Thyroid Hormone Insufficiency Causes Structural Anomalies in the Embryo of Domestic Chick, Gallus domesticus.

Thyroid hormone (TH) is essential for growth and development, yet its specific role during embryogenesis remains incompletely understood. This study investigates the impact of TH deficiency, induced by thiourea, a known inhibitor of thyroid peroxidase (TPO), on the development of domestic chicks. Thiourea was administered before thyroid gland formation, and its presence in treated embryos was confirmed through liquid chromatography-mass spectrometry. In silico docking revealed a strong interaction between thiourea and the CCP-like domain of TPO, which was corroborated by TPO activity assays showing reduced enzyme function. This reduction in enzyme activity led to lower embryonic TH levels and increased thyroid-stimulating hormone (TSH) secretion. Morphological analysis of newly hatched chicks revealed significant structural anomalies, particularly in lateral plate mesoderm-derived structures, including omphalocele, limb deformities, anophthalmia and craniofacial defects. Alcian blue and Alizarin red staining demonstrated reduced ossification in ribs and forelimbs, while histological analysis showed incomplete abdominal wall closure and abnormal vertebral column development. Haematological profiling of TH-deficient newly hatched chicks revealed significantly lower blood cell counts, highlighting TH's critical role in haematopoiesis. These findings emphasise the multifaceted role of TH in embryonic development, with potential implications for understanding congenital hypothyroidism and its developmental impacts, especially in regions with limited healthcare access.

Preoperative abaloparatide plus zoledronate treatment accelerates femoral bone healing in rats following osteotomy.

Purpose: We investigated the potential efficacy of abaloparatide (Abalo) and zoledronate (ZA) combination therapy for accelerating femoral union in a rat osteotomy model. Methods: Nine-week-old male Sprague-Dawley rats were randomly divided into four groups (n = 14 per group): control, abaloparatide (Abalo, 30μg/kg via subcutaneous injection [s.c.] 5 times per week for 6weeks), zoledronate (ZA; 0.1mg/kg via s.c., single dose), and Abalo + ZA. Rats were then subjected to unilateral osteotomy of the femoral shaft, followed by osteosynthesis with intramedullary nailing to establish bone healing models. At 2 and 4weeks after osteotomy, both femurs were removed from seven rats per group for soft X-ray imaging to evaluate bone union and microcomputed tomography (micro-CT) for bone morphometric evaluation. Blood samples were collected from all rats every 2week starting 6weeks pre- to 4weeks postosteotomy. Toluidine blue staining was used for histopathological evaluation of the undecalcified specimens. Results: Soft X-ray imaging revealed accelerated callus formation, callus maturation, and fracture line closure in Abalo and Abalo + ZA groups compared to ZA and control groups. Micro-CT demonstrated greater cortical bone and trabecular bone to total volume ratios in contralateral (left) femurs of the Abalo + ZA group compared to the Abalo group. Both trabecular and cortical bone mineral densities were also greater in contralateral femurs of the Abalo + ZA group compared to Abalo and ZA groups. Conclusion: These findings suggest substantial additive or synergistic efficacy of abaloparatide plus zoledronate combination therapy for accelerating bone healing following osteotomy and for maintaining normal bone health.

Effect of Danhong Injection on Articular Cartilage Degeneration in Rabbits with Knee Osteoarthritis

The aim of this research was to investigate the effect of Danhong injection nanoparticle (DIN) plus warm acupuncture (WA) on articular cartilage degeneration in knee osteoarthritis (KOA) rabbits. First, the KOA model and DIN were made. After success, the rabbits were randomly grouped: according to the treatment methods: controls, DIN, WA, and DIN+WA groups, with 10 rabbits in each group. The controls did not receive any treatment, and the other groups received corresponding treatment intervention, respectively. Through treatment, articular cartilage samples were collected. The bone morphology, LequesneMG score, Mankin score were analyzed. The safranin-fast green, toluidine blue, Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick End Labeling (TUNEL), and immunohistochemical staining were observed. The levels of apoptosis-related factors were determined. The results showed that the KOA rabbit model and DIN were successfully constructed. Compared with other groups, the LequesneMG and Mankin scores of DIN+WA group decreased markedly (P &lt; 0.05). Safranin-fast green and toluidine blue staining showed that the number of chondrocytes in DIN+WA group was markedly increased, which was more as against other groups. TUNEL staining revealed that the number of apoptotic chondrocytes was markedly reduced, and the apoptotic index was markedly lower in DIN+WA group as against other three groups (P &lt; 0.05). Compared with other groups, Bax and Caspase-3 were markedly decreased, and Bcl-2 was markedly raised in DIN+WA group (P &lt; 0.05). According to the review, the combined application of DIN and WA can promote the growth of chondrocytes, inhibit the apoptosis of chondrocytes, and effectively improve the degeneration of knee articular cartilage, which has application value.

Melatonin alleviates heme-induced ferroptosis via activating the Nrf2/HO-1 pathway in neurons.

Ferroptosis of neurons is a significant cause of brain injury following intracerebral hemorrhage (ICH). As an iron-containing compound in hemoglobin, heme contributes to nerve injury post-ICH. Melatonin has been shown to mitigate the effects of ICH, yet its specific functions remain largely elusive. In this study, we aimed to explore the roles and mechanisms of melatonin in heme-induced ferroptosis subsequent to ICH. C57BL/6 mice were intracranially injected with heme and then treated with melatonin. Behavior tests [modified neurological severity score (mNSS), forelimb placing, and corner turn tests], H&E staining, Nissl staining, and Prussian blue staining were used to evaluate mouse brain tissue injury. In vitro, HT-22 cells were stimulated with heme and cell viability was determined by crystal violet staining. The iron contents were determined in heme-treated brains and cells, and the levels of 4-hydroxynonenal (4-HNE) and malonaldehyde (MDA) were assessed by ELISA. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to investigate the mRNA levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). Immunoblotting was used to analyze the protein expression of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), Nrf2, and HO-1. Finally, small interfering RNA (siRNA) was used to knock down Nrf2 in HT-22 cells. Melatonin treatment alleviated heme-induced injuries to neural function, as indicated by improved behavior in the mice. Moreover, melatonin decreased cell death and iron concentrations, increased MDA and 4-HNE levels, and reversed the decreases in GPX4, SLC7A11, Nrf2, and HO-1 induced by heme in vitro and in vivo. These results indicated that melatonin could improve the ferroptosis induced by heme. In addition, we found that Nrf2 knockdown attenuated the therapeutic effect of melatonin on neuronal ferroptosis induced by heme. In general, melatonin alleviates heme-induced ferroptosis by activating the Nrf2/HO-1 pathway, which implies that melatonin is a promising treatment for ferroptosis in ICH.

Comparison of the structural integrity and quality of corneal endothelium stored in organ culture storage medium versus Eusol-C.

In this experimental study, we compared the structural integrity and cell quality of corneal endothelium stored in organ culture medium (OCS) and Eusol-C. The experiment included rabbit and human cornea experiments in vitro. Thirty rabbit corneas and thirty-two human corneas were collected and divided into two groups. All right corneas were allocated in experiment group and left corneas were placed in control group. The corneas in experimental group were stored in OCS at 34°C, and the corneas in control group were stored in Eusol-C at 4°C for 7, 14, 21, 28, and 35days, respectively. Endothelial cell morphology, cell count, and trypan blue staining for viability were assessed before storage (Day 0) and at days 7, 14, 21, 28 and 35. The structural integrity of human corneal endothelial cell was analyzed using immunohistochemistry. The samples of storage solution for microbial culture were collected on the third day and at the end of storage. The results show that no bacterial and fungal infections were found in both groups. After 14days of storage, the morphology of endothelial cell was better in the experimental group than in the control group. The endothelial cell stored in OCS were better than those stored in Eusol-C at the end of storage times, except human cornea 14days storage group. The ZO-1 protein staining showed the typical polygonal morphology of endothelial cell stored in the OCS. Corneal endothelial cells stored in the OCS had better quality up to 28days. It can be applied to Chinese eye banks as a method of corneal preservation.

Development of a mouse model of chronic ventral spinal cord compression: Neurobehavioral, radiological, and pathological changes.

The main objective of this study was to establish a mouse model of spinal ligament ossification to simulate the chronic spinal cord compression observed in patients with ossification of the posterior longitudinal ligament (OPLL). The study also aimed to examine the mice's neurobiological, radiological, and pathological changes. In the previous study, a genetically modified mouse strain was created using Crispr-Cas9 technology, namely, Enpp1 flox/flox /EIIa-Cre (C57/B6 background), to establish the OPLL model. Wild-type (WT) mice without compression were used as controls. Functional deficits were evaluated through motor score assessment, inclined plate testing, and gait analysis. The extent of compression was determined using CT imaging. Hematoxylin and eosin staining, luxol fast blue staining, TUNEL assay, immunofluorescence staining, qPCR, and Western blotting were performed to evaluate levels of apoptosis, inflammation, vascularization, and demyelination in the study. The results demonstrated a gradual deterioration of compression in the Enpp1 flox/flox /EIIa-Cre mice group as they aged. The progression rate was more rapid between 12 and 20 weeks, followed by a gradual stabilization between 20 and 28 weeks. The scores for spinal cord function and strength, assessed using the Basso Mouse Scale and inclined plate test, showed a significant decline. Gait analysis revealed a noticeable reduction in fore and hind stride lengths, stride width, and toe spread. Chronic spinal cord compression resulted in neuronal damage and activated astrocytes and microglia in the gray matter and anterior horn. Progressive posterior cervical compression impeded blood supply, leading to inflammation and Fas-mediated neuronal apoptosis. The activation of Bcl2 and Caspase 3 was associated with the development of progressive neurological deficits (p < 0.05). The study presents a validated model of chronic spinal cord compression, enabling researchers to explore clinically relevant therapeutic approaches for OPLL.

EphrinB2-mediated chondrocyte autophagy induces post-traumatic arthritis via rupture of cartilage homeostasis.

EphrinB2, a member of the Ephrin family, has been linked to several orthopaedic conditions. Nevertheless, the correlation between ephrinB2 and post-traumatic arthritis (PTOA) remains unclear. Human PTOA cartilage from human and mouse knee joints was systematically analysed to investigate the relationship between EphrinB2 and PTOA using SO-FG and toluidine blue staining, micro-CT, histomorphometry, immunohistochemistry, immunofluorescence, lentiviral articular injection and insitu end labeling (TUNEL) assays. EphrinB2 expression was significantly downregulated in PTOA chondrocytes. Blocking EphrinB2 increased the breakdown of cartilage matrix in mice with PTOA via reducing the process of chondrocyte autophagy. The presence of severe cartilage damage was evident, as indicated by a considerable decrease in both cartilage thickness and area, accompanied by an increase in chondrocyte death. Altogether, EphrinB2 is required for the maintenance of cartilage homeostasis in post-traumatic arthritis, and EphrinB2 ablation is associated with accelerated chondrocyte matrix degeneration, finally causing damage to the articular cartilage.

Clinicopathological risk factors of oral second primary tumours

BackgroundOral second primary tumours (SPTs) have a poor prognosis due to late-stage diagnosis. This study evaluates the demographic and clinicopathological risk predictors of SPTs. MethodsPatients with oral squamous cell carcinoma, carcinoma in situ, or severe dysplasia were accrued into the Oral Cancer Prediction Longitudinal study within one year post-curative treatment. Data on demographics, risk habits, and primary tumour characteristics were collected. Clinical follow-up included assessing the presence of second oral premalignant lesions (SOPLs), clinicopathological features, and the results from toluidine blue staining and fluorescence visualization. ResultsAmong 296 patients, 23 (8 %) developed SPTs. Older age at primary cancer diagnosis (P = 0.008) and a history of chewing tobacco or betel nut (P = 0.043) increased the risk of SPTs. Patients with primary tumours located at low-risk sites had an increased risk of SPTs (P = 0.004), which often presented at high-risk sites. The presence of SOPLs (P < 0.001), and multiple lesions (P = 0.017) significantly increased the risk of SPTs. Positive toluidine blue staining indicated a trend toward higher risk of SPTs, whereas fluorescence visualization did not. The median time to SPT diagnosis was 3.25 years post-treatment. ConclusionsIdentifying second or multiple oral premalignant lesions is critical for predicting the risk of SPTs regardless of their clinical or histological characteristics. Routine biopsy of these lesions should be prioritized to ensure timely diagnosis. Incorporating these risk predictors into clinical follow-up can enhance early cancer detection and improve patient outcomes.

Clinicomycological Profile of Dermatophytosis in Tripura – An Institution Based Cross Sectional Study

Abstract Background: In a small state like Tripura, a surge in prevalence of superficial dermatophytosis has been observed from April to September among the population of both tribal and non-tribal groups having different lifestyle, different food habit, and different socioeconomic status. Studies from different region reveal varying pattern of etiological distribution of the disease. But till date no research study has been initiated in Tripura on clinicomycological profile of dermatophytosis. Objectives: This study was undertaken with the objectives to estimate the proportion of different fungal species associated with dermatophytosis among the patients attending Dermatology Outpatient Department of a Tertiary care hospital of Tripura and also to determine possible association of different clinical parameters with fungal species if any. Materials and Methods: A total of three hundred and ninety-one (391) new cases of dermatophytosis attending Dermatology Outpatient Department of a Tertiary care teaching hospital of Tripura were included in this study from June, 2021 to May, 2022. All the specimens were assessed by direct microscopic examination and in vitro culture. Fungi were identified on the basis of their macroscopic and microscopic features with the help of lactophenol cotton blue staining and urease test. Results: There was a male preponderance among cases and maximum patients belonged to 21-40 years age group. Trichophyton schoenleinii was the most common fungal species (52.1%) grown in culture followed by Trichophyton rubrum (26.9%). Conclusion: In contrast to previous study results from different parts of India, Trichophyton schoenleinii was the most common isolated fungal species from this part of northeast India, and Tinea corporis (41.3%) was the most prominent clinical manifestation of dermatophytosis.

EVALUATION OF WOUND HEALING AND EPITHELIALIZATION USING TOLUIDINE BLUE DYE INDICATOR IN DIODE LASER FRENECTOMY WITH AND WITHOUT SURGICAL MICROSCOPE

Objective: The study aimed to evaluate the wound healing and epithelialization in diode laser frenectomy using toluidine blue dye as an indicator, comparing outcomes between procedures performed with and without the use of a surgical microscope at 1- and 2-weeks post-operatIvely. Materials and Methods: This controlled clinical trial involved patients undergoing diode laser frenectomy, divided into two groups: one group underwent the procedure with the aid of a surgical microscope, while the other group did not use a microscope. Wound healing and epithelialization were assessed using toluidine blue dye at 1 and 2 weeks following the procedure. Results: The group that received diode laser frenectomy with the use of a surgical microscope demonstrated significantly faster and more complete wound healing and epithelialization at both 1 and 2 weeks compared to the group that did not use the surgical microscope. The toluidine blue stain uptake showed similar results. Conclusion: The use of a surgical microscope in diode laser frenectomy appears to enhance wound healing and epithelialization, as evidenced by the results at both 1 and 2 weeks. This suggests that incorporating a surgical microscope into the procedure may improve clinical outcomes and reduce recovery time. Further research is needed to confirm these findings across larger and more diverse patient populations.

Endogenous Adult Telomerase Positive Stem Cells Increase in Equine Peripheral Blood Following Exercise

Stout, et al., 2007 reported a base-line level of primitive endogenous adult Telomerase Positive Stem Cells (aTPSCs) circulating within adult porcine peripheral blood which increased dramatically, i.e., 23.5-fold, after just 90 minutes of trauma. Ongoing studies from our laboratory have shown the presence of similar primitive stem cells in adult equine blood and that the base-line levels of stem cells varied according to the particular breed of horse. The current study was undertaken to test the hypothesis that aTPSCs would increase in number in the peripheral blood following stress in the same horse. Stress in this study was defined as moderate exercise, i.e., 10 minutes of cantering. Blood withdrawal followed the guidelines of Fort Valley State University’s IACUC. Adult horses had their blood withdrawn immediately prior to and immediately after exercising. The blood was obtained by venipuncture. Isolated aTPSCs were assayed by trypan blue staining, CEA-CAM-1 for totipotent stem cells and SSEA-4 for pluripotent stem cells. This study demonstrates that moderate exercise will increase the level of these primitive pluripotent stem cells in the blood of adult equines. Studies are ongoing to address their functional significance during injury and repair.