Histopathological Changes Research Articles

The protective efficacy of omega-3 polyunsaturated fatty acids on oxidative stress, inflammation, neurotransmitter perturbations, and apoptosis induced by monosodium glutamate in the brain of male rats.

Exaggerated neuronal excitation by glutamate is a well-known cause of excitotoxicity, a key factor in numerous neurodegenerative disorders. This study examined the neurotoxic effect of monosodium glutamate (MSG) in the brain cortex of rats and focused on assessing the potential neuroprotective effects of omega-3 polyunsaturated fatty acids (ω-3 PUFAs). Four groups of adult male rats (n = 10) were assigned as follows; normal control, ω-3 PUFAs (400mg/kg) alone, MSG (4mg/g) alone, and MSG plus ω-3 PUFAs (4mg/g MSG plus 400mg/kg ω-3 PUFAs). Biochemical analysis, immunohistochemical, and histological examinations were conducted upon completion of the treatment protocol. Results revealed that MSG significantly increased malondialdehyde, nitric oxide, tumor necrosis factor-α, interleukin 1β, acetylcholinesterase, monoamine oxidase, and caspase-3. However, the MSG-treated group showed a decline in reduced glutathione, catalase, superoxide dismutase, dopamine, and serotonin. In addition, MSG caused histopathological changes in the cortical region which support the biochemical and immunohistochemical analysis. Supplementation of ω-3 PUFAs greatly improved the biochemical, immunohistochemical, and histopathological alterations induced by MSG administration in the brain cortex. Together, these findings revealed a neuroprotective effect of ω-3 PUFAs against MSG-induced toxicity in the brain cortex by attenuating oxidative damage, inflammation, neurochemical perturbations, and apoptosis.

Effects of Tasimelteon Treatment on Traumatic Brain Injury Through NRF-2/HO-1 and RIPK1/RIPK3/MLKL Pathways in Rats.

Secondary brain damageafter traumatic brain injury (TBI) involves oxidative stress, neuroinflammation, apoptosis, and necroptosis and can be reversed by understanding these molecular pathways. The objective of this study was to examine the impact of tasimelteon (Tasi) administration on brain injury through the nuclear factor erythroid 2-related factor 2 (NRF-2)/heme oxygenase-1 (HO-1) and receptor-interacting protein kinase 1 (RIPK1)/receptor-interacting protein kinase 3 (RIPK3)/mixed lineage kinase domain-like (MLKL) pathways in rats with TBI. Thirty-two male Wistar albino rats weighing 300-350g were randomly divided into four groups: the control group, trauma group, Tasi-1 group (trauma + 1mg/kg Tasi intraperitoneally), and Tasi-10 group (trauma + 10mg/kg Tasi intraperitoneally). At the end of the experimental phase, after sacrifice, blood samples and brain tissue were collected for biochemical, histopathological, immunohistochemical, and genetic analyses. Tasi increased the total antioxidant status and decreased the total oxidant status and oxidative stress index. In addition, Tasi caused histopathological changes characterized by a markedly reduced hemorrhage area in the Tasi-1 group. Normal brain and meningeal structure was observed in rats in the Tasi-10 group. Immunohistochemical analysis indicated that Tasi also decreased the expression of interferon-gamma, caspase-3, and tumor necrosis factor-alpha in the brain tissue. Although NRF-2 and HO-1 expression decreased, RIPK1/RIPK3/MLKL gene expression increased due to trauma. However, Tasi treatment reversed all these findings. Tasi protected against brain injury through the NRF-2/HO-1 and RIPK1/RIPK3/MLKL pathways in rats with TBI.

IFN-β production induced by PRRSV is affected by GP3 quantity control and CLND4 interaction

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most harmful pathogens in the swine industry. Our previous studies demonstrated that the small extracellular domain (ECL2) of CLDN4 effectively blocks PRRSV infection. In this study, we explored the in vivo administration of swine ECL2 (sECL2) and found that it blocked HP-PRRSV infection and alleviated histopathological changes in organs. Notably, sECL2 stimulated cytokine production in the lungs. We observed that CLDN4 upregulated the expression of IFN-β at low doses of GP3. While high doses of GP3 inhibited the activity of the IFN-β promotor, regardless of whether CLDN4 was present. GP3 also downregulated IFN-β by decreasing the phosphorylation of TBK1 and IRF3. These findings highlight functional differences in GP3 under quantity control, which account for the variations in IFN-β induction during the early and late stages of infection. Our results indicate that sECL2 is a promising candidate drug for developing treatments to control PRRS.

Prepubertal phthalate exposure can cause histopathological alterations, DNA methylation and histone acetylation changes in rat brain.

Di-2-(ethylhexyl)phthalate (DEHP) is a phthalate derivative used extensively in a wide range of materials, such as medical devices, toys, cosmetics, and personal care products. Many mechanisms, including epigenetics, may be involved in the effects of phthalates on brain development. In this study, Sprague-Dawley male rats were obtained 21-23days after their birth (post-weaning) and were exposed to DEHP during the prepubertal period with low-dose DEHP (DEHP-L, 30mg/kg/day) and high-dose DEHP (DEHP-H, 60mg/kg/day, 37days) until the end of adolescence (PND 60). The rats in the study groups were sacrificed during adulthood, and histopathological changes, epigenetic changes, and oxidative stress parameters were evaluated in brain tissues. Histopathological findings indicating the presence of deterioration in brain tissue morphology were obtained, more prominently in the DEHP-H group. Examining the hippocampus under the light microscope, pyramidal neuron loss was detected only in CA1 of the DEHP-L group, while in DEHP-H rats, pyramidal neuron losses were detected in the CA1, CA2, and CA3 regions. No significant change was observed in brain lipid peroxidation levels with DEHP compared to control. Significant increases in total glutathione (GSH) in both dose groups were considered to be an adaptive response to DEHP-induced oxidative stress. The decrease in DNA methylation in the brain, although not statistically significant, and the increase in histone modification showed that exposure to DEHP may cause epigenetic changes in the brain and these epigenetic changes may also take place as one of the mechanisms underlying the damage observed in the brain. The results suggest that DEHP exposure during early development may have a significant effect on brain development.

Impact of Smoking on Cervical Histopathological Changes in High-Risk HPV-Positive Women: A Matched Case–Control Study

Impact of Smoking on Cervical Histopathological Changes in High-Risk HPV-Positive Women: A Matched Case–Control Study

Astaxanthin Alleviates Lung Injury by Regulating Oxidative Stress, Inflammatory Response, P2X7 Receptor, NF-κB, Bcl-2, and Caspase-3 in LPS-Induced Endotoxemia.

Sepsis remains the leading cause of multiple-organ injury due to endotoxemia. Astaxanthin (ASTA), widely used in marine aquaculture, has an extraordinary potential for antioxidant and anti-inflammatory activity. Purinergic receptor (e.g., P2X7R) activation is a powerful signaling in the modulation of inflammation. The effect of ASTA was investigated on the regulation of oxidative stress, inflammatory response, apoptotic mediators, and P2X7R expression in the lung injury during lipopolysaccharide (LPS)-induced endotoxemia. Twenty-four rats were blocked into four groups as Control, LPS, ASTA, and LPS + ASTA. LPS was administered by intraperitoneal injection and ASTA by gavage. Blood and lung samples were taken 6 h after the administrations. The methods were ELISA, western blotting, histopathology, and immunohistochemistry. Sepsis was confirmed by the elevations of IL-1β, IL-6, IL-10, and TNF-α levels in bloodstream. Lung injury was determined by histopathological changes. There were increased P2X7R expression, malondialdehyde (MDA), IL-1β, TNF-α, nuclear factor kappa B (NF-κB), and Caspase-3 and decreased B-cell lymphoma 2 (Bcl-2) and glutathione (GSH) in the septic lung tissue (p < 0.05). ASTA treatment improved MDA, GSH, IL-1β, TNF-α, P2X7R, NF-κB, Caspase-3, and Bcl-2 levels and reduced P2X7R immunoreactivity and histological abnormalities in the lung (p < 0.05). The production of pro-inflammatory cytokines, oxidative stress, P2X7R expression, and apoptotic mediators in the lung is associated with LPS-induced endotoxemia. The ASTA administration appears to regulate the expressions of P2X7R, NF-κB, Bcl-2, and Caspase-3 improving the antioxidative and anti-inflammatory response of the lung tissue in sepsis, invivo.

Angiotensin 1-7 Attenuates the Development of Ischemia-Reperfusion-Induced Arrhythmia in Rats: Electrophysiology, Molecular, and Immunohistochemical Study.

Arrhythmia is a common and serious global health problem, contributing to cardiovascular morbidity and mortality. The cardiac muscle is susceptible to ischemia-reperfusion (I/R) injury, which can lead to fatal arrhythmias during open-heart surgery. We investigated the potential prophylactic effect of angiotensin 1-7 (Ang 1-7) using an invivo rat model of I/R injury and examined the underlying mechanisms. Rats were treated with Ang 1-7 (1 mg/kg, IP) 30 min before the surgical procedures. Twenty-four rats were equally divided into four groups: sham control, sham-treated with Ang 1-7, I/R injury group, and I/R injury group treated with Ang 1-7. Invivo I/R injury was induced by clamping the left coronary artery for 30 min, followed by 1 hour of reperfusion. The I/R group showed abnormal electrophysiological changes and arrhythmic episodes during electrocardiography (ECG) recording, increased oxidative stress, downregulation of peroxisome proliferator-activated receptor gamma (PPAR-γ), and upregulation of C-X-C motif chemokine ligand 16 (CXCL16) expression in cardiac tissue, which increased cardiac NF-kB expression and IL-17 levels. Moreover, I/R injury caused significant histological disruption and increased cyclooxygenase 2 (COX-2) and heat shock protein 90 (HSP90) immunoreactions, correlating with the extent of cardiac damage. However, preoperative Ang 1-7 administration significantly improved the electrophysiological, biochemical, and histopathological changes induced by I/R injury. This study demonstrated that Ang 1-7 exerted protective anti-arrhythmic, anti-inflammatory, and pro-healing effects by upregulating PPAR-γ and downregulating CXCL16, IL-17, and NF-kB pathways, suggesting it is a promising cardioprotective agent for preventing arrhythmias induced by I/R injury.

Modulation of placental angiogenesis by metformin in a rat model of gestational diabetes.

Gestational diabetes mellitus (GDM) significantly disrupts placental structure and function, leading to complications such as intrauterine growth restriction (IUGR) and preeclampsia. This study aimed to investigate the effects of GDM on placental histology, angiogenesis, and oxidative stress, as well as evaluate metformin's protective role in mitigating these changes. A total of 60 pregnant Sprague-Dawley rats were divided into four groups: control, metformin-treated, GDM, and GDM with metformin. GDM was induced using streptozotocin (STZ) at 40mg/kg, and metformin was administered at 200mg/kg from gestational day (GD) 4 to GD17. Blood glucose and insulin levels were assessed, and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) was calculated. Placentae were weighed and subjected to histological, immunohistochemical, and molecular analyses, focusing on key angiogenesis markers (VEGF, VEGFR, CD31, KLF2) and oxidative stress indicators (MDA, eNOS). GDM increased placental weight, angiogenesis (elevated VEGF, VEGFR, CD31), and oxidative stress (elevated MDA, eNOS). Histopathological changes included villous edema, membrane rupture, and hemosiderin deposition. Metformin treatment reduced placental weight; normalized VEGF, KLF2, and PlGF expression; and improved placental architecture. Additionally, oxidative stress was significantly reduced in metformin-treated GDM rats. In conclusion, GDM induces placental abnormalities, promoting excessive angiogenesis and oxidative stress, potentially leading to IUGR and other complications. Metformin showed protective effects by reducing placental overgrowth and restoring vascular and oxidative balance. These findings suggest that metformin may play a therapeutic role in improving placental health in GDM pregnancies, warranting further investigation into its long-term effects on fetal development and maternal health.

Exploration of the Combined Mechanism of Direct and Indirect Effects of Paeoniflorin in the Treatment of Cholestasis.

Cholestasis is a multifactorial hepatobiliary disorder, characterized by obstruction of bile flow and accumulation of bile, which in turn causes damage to liver cells and other tissues. In severe cases, it can result in the development of life-threatening conditions, including cirrhosis and liver cancer. Paeoniflorin (PF) has been demonstrated to possess favourable therapeutic potential for the treatment of cholestasis. The objective of this research was to examine the molecular mechanism of PF in the treatment of ANIT-induced cholestasis and to propose novel avenues for further research on the pharmacological effects of PF. In vivo and in vitro models of cholestasis were developed. The histopathological changes in the bile ducts and liver were evaluated through the use of hematoxylin and eosin (HE) staining. The extent of apoptosis was evaluated through the use of immunofluorescence (IF), immunoblotting (WB), and electron microscopy. The JNK signalling pathway was identified as the direct mechanism of action of PF through the utilisation of HuProt™ 20K chips and other technologies. The present study demonstrated that PF markedly alleviated liver injury in an ANIT-induced cholestasis model. Specifically, PF was observed to attenuate cholestasis-induced liver injury by reducing the abnormal elevation of liver function indices and suppressing the expression of inflammatory mediators. Furthermore, PF exhibited anti-apoptotic properties in both in vivo and in vitro experiments, thereby mitigating cholestasis-induced hepatocyte apoptosis. These protective effects are attributable to the fact that PF exerts its action through direct interaction with the JNK pathway. It has been demonstrated that PF is capable of binding directly to MAPK8 (JNK1) and MAPK9 (JNK2), thereby inhibiting JNK activation and reducing apoptosis. With regard to the protection of bile ducts, PF may indirectly inhibit hepatocyte apoptosis by maintaining the structural integrity and tight junctions of bile duct cells. PF improved cholestasis by inhibiting hepatocyte apoptosis directly by targeting the JNK signaling pathway and indirectly inhibited hepatocyte apoptosis by improving the tight junctions of bile duct cells to regulate the bile duct microenvironment.

ANTI-TOXOPLASMA GONDII ANTIBODIES IN EGYPTIAN PATIENTS WITH SCHIZOPHRENIA

For nearly 100 years, infectious microbes dominated discourse on the causes of schizophrenia, Toxoplasma gondii is an obligate intracellular parasite and is found in 2 forms in humans. Toxoplasma gondii has been associated with several congenital CNS anomalies and more subtle delayed neurologic sequelae, The evidence of infection as a cause of schizophrenia was supported by the brain histopathological changes found in patients with schizophrenia, the risk of schizophrenia increased among individuals who were exposed in utero to elevated maternal T. gondii immunoglobin G (IgG) antibody, based on assay of archived maternal sera, was more than twice that of comparison subject, The main aim of this study was to compare the prevalence of T. gondii infection between individuals with schizophrenia and healthy controls in Egyptian choret samples. 100 patients with schizophrenia, and 100 healthy donors where screened for Toxoplasma gondii IgG and IgM antibodies in serum by ELISA. A significant increase in the incidence of Toxoplasma gondii IgG positive antibody in patient group compared to control group (P 0.004)

Correlations between postmortem quantitative MRI parameters and demyelination, axonal loss and gliosis in multiple sclerosis: A systematic review and meta-analysis.

Magnetic resonance imaging (MRI) is frequently used to monitor disease progression in multiple sclerosis (MS). This study aims to systematically evaluate the correlation between MRI measures and histopathological changes, including demyelination, axonal loss, and gliosis, in the central nervous system of MS patients. We systematically reviewed post-mortem histological studies evaluating myelin density, axonal loss, and gliosis using quantitative imaging in MS. Relevant studies were identified through searches in PubMed, EMBASE, and Web of Science. A total of 38 studies involving 1782 regions of interest from 229 subjects were included. Pooled random-effects models were used to calculate the correlation between demyelination, axonal loss, gliosis, and various MRI parameters, including magnetization transfer ratio (MTR), T1 and T2 relaxation times, myelin water fraction (MWF), proton density (PD), and diffusivities. Pair-wise analyses compared results between lesioned and non-lesioned tissues. Our results demonstrated moderate to strong correlations between MRI parameters and myelin density in MS, with correlation coefficients: T1 (0.72), T2 (0.72), MTR (-0.73), FA (-0.73), RD (0.70), MD (0.70), MWF (-0.82), and PD (0.73). Interestingly, stronger correlations were found in lesioned tissues compared to non-lesioned tissues (P < 0.001). Moderate correlations were found between MRI parameters and axonal loss and gliosis. Our study reveals significant correlations between MRI techniques and histological assessments of myelin, axonal damage, and gliosis in MS. MRI metrics exhibited a more robust association with demyelination in lesioned areas than in non-lesioned brain tissue, highlighting the pronounced degree of myelin degradation in MS lesions. Further investigation is warranted to corroborate these results and refine MRI-based monitoring of MS pathology.

Injection of hyaluronic acid versus platelet rich plasma for treatment of vulvovaginal atrophy in post-menopausal females.

Female sexual dysfunction is highly prevalent among postmenopausal females approaching 50%, with vulvovaginal atrophy (VVA) being a cardinal sign. For decades, hormone replacement therapy was the only solution to relieve symptoms associated with this atrophy. However, it was limited by its serious side effects, raising the need for new treatment strategies. This study aims to compare the efficacy and safety of injection of hyaluronic acid (HA) versus platelet rich plasma (PRP) in post-menopausal females presented with VVA to improve female sexual dysfunction. Twenty post-menopausal females presented with VVA were randomly divided into two groups, 10 patients in each group. Both groups received three sessions of injections into the vulva and vagina, one month apart. Group I was injected with non cross linked HA while Group II received PRP injection. Subjective assessment was carried out through female sexual function index questionnaire and global aesthetic improvement scale.While objective assessment was carried out by measuring the labia majora length and reviewing the histopathological changes occurring in the vulva through skin biopsies before and after treatment. The change in vaginal thickness was estimated by transvaginal ultrasound. Results showed that both HA as well as PRP were effective in the treatment of post-menopausal vulvovaginal atrophy. However, HA showed more significant improvement in female sexual dysfunction. There were also higher values in vaginal wall thickness as measured by transvaginal ultrasound in favor of HA injected group. Histopathological assessment showed more collagen deposition in papillary dermis in HA treated group. No complications were reported in both groups.

In vivo toxic and lethal cardiorespiratory effects of a synthetic quaternary ammonium salt derivative of haloperidol in mice.

To investigate the toxicity of N-n-butyl haloperidol iodide (F2), a quaternary ammonium salt derivative of haloperidol, in mice for potential therapeutic purposes. The acute median lethal dose (LD50) of F2 was determined using the Bliss method following intravenous administration in mice. Routine surface electrocardiograms (ECGs) and arterial blood pressures (aBPs) were recorded under general anesthesia in untreated and pharmacologically vagotomized mice injected with F2. Sublethal doses of F2 were tested for their effects on aBP, heart rate, and biochemical parameters such as lactate dehydrogenase (LDH), blood urea nitrogen (BUN), and serum lactate levels. Histopathological changes in the heart, lungs, liver, and kidneys were evaluated after F2 administration. The acute LD50 of F2 was determined to be 5.11 mg/kg. A 10 mg/kg dose of F2 caused severe hypotension, second-degree atrioventricular block, progressive prolongation of Pmurr intervals, and death due to cardiac asystole. Similar ECG and aBP changes were observed in atropine-pretreated mice, indicating that cholinergic effects do not play a major role in F2-induced toxicity. Sublethal doses of F2 (1.2 and 2.4 mg/kg) caused dose-dependent decreases in aBP and increases in heart rate. F2 induced significant, dose-dependent increases in LDH, BUN, and serum lactate levels. Histopathological analysis revealed acute lung lesions at 10 mg/kg, with no significant changes observed in the heart, liver, or kidneys. Acute intravenous injection of F2 exhibits dose-dependent cardiopulmonary toxicity, characterized by severe hypotension, arrhythmias, and biochemical changes. These findings highlight the potential risks of F2 and the need for further evaluation of its safety profile for therapeutic use.

Mechanistic insights of methylcinnamate in improving oxidative stress and inflammation in acetaminophen-induced hepatotoxic mice by upregulating Nrf2 pathway.

Methylcinnamate (MC), a safe flavoring agent naturally found in Occimum basilicum L. is reported to have an anti-inflammatory responses in various disease models. Acetaminophen (APAP) toxicity is a significant contributor to acute liver injury, which leads to oxidative stress and inflammation. The transcriptional factor nuclear factor erythroid 2-related factor 2 (Nrf2) regulated the cellular defense mechanisms aid to antioxidant response facilitation and reduction in inflammation against various disorders. This study evaluated the protective effects of MC in APAP-induced hepatotoxicity in mice and its anti-oxidant, anti-inflammatory, and Nrf2 mechanisms were studied. In-vitro 2,2-diphenyl-1-picrylhydrazyl assay showed the antioxidant capacity of MC. Mice were pretreated with MC (25, 50, 75, and 100mg/kg) orally for 7 days. After a fasting period of 16h, hepatotoxicity was induced by injecting APAP 300mg/kg intraperitoneal on day 7. Liver profile, oxidative test, and histopathological changes were studied. Gene expression of interlukin-1β (IL-1β), interlukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), cytochrome P450 2E1 (CYP2E1), Nrf2, and NAD(P)H dehydrogenase (quinone) 1 (NQO-1) were estimated by real time quantitative polymerase chain reaction (RT-qPCR). IL-1β, IL-6, and TNF-α concentrations were also analyzed by enzyme-linked immunosorbent assay (ELISA). The MC treatment showed a notable reduction in alanine transaminase, aspartate aminotransferase and alkaline phosphatase activities, and total bilirubin level of serum. Moreover, MC significantly attenuated oxidative stress by rising the antioxidant enzymes catalase, glutathione, and superoxide dismutase and reducing the malondialdehyde and nitric oxide levels in the liver. Furthermore, MC successfully mitigated the levels of IL-1β, IL-6, and TNF-α, which were estimated through RT-qPCR and ELISA. The RT-qPCR revealed a CYP2E1 enzyme inhibition and significant upregulation of hepatic Nrf2 and NQO-1 levels after MC therapy. Histopathological analysis showed improvement in liver injury within the MC treatment groups. It was concluded from this study that pretreatment of MC had successfully protected the liver through anti-inflammatory, anti-oxidant activity upon subsequent activation of Nrf2.

Potential of Lambda-Cyhalothrin for Controlling the Black Armyworm Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae): Toxicity and Midgut Histopathological Effects.

Caterpillars of the genus Spodoptera are the main pests in soybean and cotton crops and Spodoptera cosmioides causes more severe losses than other caterpillars in these agricultural crops. However, there are few recommended insecticides for controlling this pest. Lambda-cyhalothrin is a pyrethroid used to control a wide spectrum of arthropods including lepidopterans. Therefore, the objective of this study was to evaluate the potential of lambda-cyhalothrin for the control of S. cosmioides. Specifically, toxicity and histopathological changes in the midgut were evaluated. The effectiveness of the insecticide was determined by estimating the different lethal concentrations (LCs) ​​in the laboratory upon S. cosmioides. Lambda-cyhalothrin was found toxic to S. cosmioides (LC50 = 23.03mg L-1 and LC90 = 174.8mg L-1), with the survival reduced from 83.33% in the control to 37.89%, 16.66%, 0%, and 0% after 72h of exposure to the LC25, LC50, LC75, and LC90 of lambda-cyhalothrin, respectively. Histopathological studies revealed that lambda-cyhalothrin caused damage to midgut cells, including epithelial disorganization, increased cytoplasmic vacuolization, brush border degeneration, nuclear chromatin condensation, and cell fragmentation, indicating cell death by apoptosis. It was concluded that lambda-cyhalothrin, a neurotoxic insecticide, caused damage to the midgut of S. cosmioides, compromising its physiology and indicating that it has potential to be used to control this pest.

Gabexate mesylate thermo-sensitive in-situ gel is effective for treating grade-III pancreatic trauma in beagle dogs guided by contrast-enhanced ultrasound.

This study evaluates the efficacy of gabexate mesylate thermosensitive in-situ gel (GMTI) in the treatment of beagle grade III pancreatic trauma (PT) with the assistance of contrast-enhanced ultrasound (CEUS) and investigates its mechanism of action. A grade III PT model consisting of 15 beagle dogs with severed main pancreatic ducts was created and treated with cephalic vein injection of gabexate mesylate (GM) (1.54 mL/10 kg, TID) and peripancreatic injection of GMTI (4.63 mL/10 kg, QD) guided by CEUS within 24 h post-surgery. Ascites and serum levels of amylase (AMY), lipase (LPS), C-reactive protein (CRP), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and urinary trypsinogen activating peptide (TAP) were detected by ELISA. Histopathological changes in the canine pancreas were observed by Hematoxylin and Eosin staining. CEUS accurately displayed pancreatic lesions and guided catheterisation. Compared to the control group, the ascites was significantly reduced after treatment (p < 0.01). AMY and LPS ascites significantly decreased on post-operative 1st and 2nd day (p < 0.01). The levels of AMY, LPS, CRP, IL-6, and TNF-α in serum were decreased (p < 0.05 or p < 0.01). Urinary TAP was decreased 1 and 2 days after treatment (p < 0.05 or p < 0.01, respectively). In the control group, pancreatic tissue necrosis was evident in the wound area. Normal glandular cell structures and fibrous tissue hyperplasia were observed in the wound area after GMTI treatment. The GMTI group performed better than the GM group in improving pancreatic histology and reducing AMY levels in the early post-operative period. Guided by CEUS, daily peripancreatic injections of GMTI in Beagles effectively inhibit pancreatic enzyme activity and aid in the adjuvant treatment of pancreatic trauma.

GABA mitigates mitochondrial apoptosis induced by high temperature stress in the Pacific oyster (Crassostrea gigas).

High temperature is a critical environmental factor leading to mass mortality in oyster aquaculture in China. Recent advancements highlight the physiological regulation function of γ-aminobutyric acid (GABA) in the adaptation of environmental stress. This study examined the physiological responses of the Pacific oyster (Crassostrea gigas) upon high temperature exposure, focusing on the histopathological changes in gill, the GABA concentration, the mRNA expression and activities of apoptosis-related genes. Following 24h of exposure to seawater at 28°C, notable histopathological changes, including cellular swelling and vacuolization, along with an increase in TUNEL-positive cells were observed in the oyster gill, compared to the control group maintained at 18°C. Moreover, there was a significant increase in CgCaspase-3 transcripts, Caspase-3 and Caspase-9 activities in the gills, glutamate decarboxylase CgGAD transcripts in the haemocytes, and GABA concentrations in the haemolymph supernatant. Intervention with GABA markedly ameliorated these responses, including reducing the mRNA expression levels of CgBax, CgBak, CgCaspase-3, and CgCaspase-9, as well as the activities of Caspase-3/9. Furthermore, after the treatment with GABAA and GABAB receptor antagonists, the activities and expression levels of Caspase-3 and Caspase-9 significantly up-regulated under hightemperature stress. GABA treatment also significantly diminished the increased Caspase-3 activity by mitochondrial pathway apoptosis inducers. High temperature induced mitochondrial pathway apoptosis via increased caspase activities. The transcripts of CgGAD in haemocytes and GABA concentration in hemolymph supernatant also increased after high-temperature stress. GABA countered these effects through the activation of GABAA and GABAB receptors, reducing both caspase activity and expression of apoptosis-related genes.

P0029 Abnormal glycerophospholipid metabolism in CCL2+DPP4+ mesenchymal stem cells may contributes to creeping fat fibrosis in Crohn's Disease

Abstract Background Creeping fat (CF), an aberrant mesenteric adipose tissue (MAT), is characterized by abnormal lipid metabolism, inflammatory cell infiltration, and extracellular matrix (ECM) deposition in the intestines affected by Crohn's disease (CD). Our previous single-cell RNA-seq analysis identified a CCL2+DPP4+ subset of mesenchymal stem cells (MSCs) that accelerates the abnormal formation of CF. Building on these findings, this study investigates the role of CCL2+DPP4+ MSCs in CF fibrosis formation. Methods We employed H&amp;E and Masson staining to assess histopathological changes and collagen deposition. Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was conducted on CCL2+DPP4+ MSCs isolated from CF biopsies of CD patients (n=5) and MAT from controls without CD (n=5). Results We observed ECM deposition in CF and an upregulation of fibrosis-related genes (COL1A1, COL1A2, and FN1)（Figure1）. Analysis of our previous single-cell data revealed that CCL2+DPP4+ MSCs play a significant role in the abnormal accumulation of the ECM (Figure2). Lipidomic profiling revealed differential lipid levels between the CF and healthy MAT（H-MAT） groups, with downregulation of specific lipids in CCL2+DPP4+ MSCs derived from CF, including PE (20:5_16:1), PC (14:0_22:6), and PE (16:1_14:0). KEGG pathway enrichment analysis highlighted significant enrichment in Glycerophospholipid metabolism. Phospholipids (PLs), crucial for membrane structure and function, were found to be regulated by key enzymes PISD(Phosphatidylserine decarboxylase) and LPCAT3(Lysophosphatidylcholine acyltransferase 3), which convert PS（phosphatidylserine） to PE（phosphatidylethanolamine） and integrate polyunsaturated fatty acids into lysophospholipids, respectively. Quantitative real-time PCR confirmed reduced expression of these enzymes and increased fibrosis gene expression in CF-derived CCL2+DPP4+ MSCs(Figure3). Conclusion Our findings suggest that the CCL2+DPP4+ MSC subset in CF exhibits abnormal glycerophospholipid metabolism, which is associated with decreased PISD and LPCAT3 expression. This may contribute to enhanced ECM formation and promotion of CF fibrosis. Understanding these mechanisms could lead to novel therapeutic approaches for CD.

Lutein, a non-provitamin A carotenoid, reduces cisplatin-induced cardiotoxicity.

Cardiovascular complications resulting from cisplatin (CS) are a significant factor that can disrupt the treatment plan associated with this chemotherapy. This information led us to investigate the effectiveness of lutein (LT), which has antioxidant effects, in preventing CS-induced cardiotoxic effects. After 28 rats were randomly divided into four equal groups, saline (1ml/day) was administered to the control group, LT (100mg/kg/day) to the LT group, CS (10mg/kg) to the CS group, and active agents in the LT and CS groups were administered to the CS + LT group in the same dose and manner. The examinations determined that MDA, cardiac biomarkers (CK-MB, BNP, LDH, and cTn-I) levels, TNF-α and caspase-3 expressions, and apoptosis significantly increased in the CS group. In contrast, GSH, SOD, and CAT levels were decreased. In addition, histopathological changes characterized by interstitial edema, leukocyte infiltration, and vacuolar degeneration were detected in the heart tissues of this group. It was determined that LT application prevented the above-mentioned CS-induced cardiotoxic effects to a significant extent, although not completely. The findings obtained in this study show that LT may reduce CS-induced cardiac damage thanks to its ROS-reducing, anti-inflammatory, anti-apoptotic, and cytoprotective characteristics.

P1312 Clickable nanozyme enhances colonization of Saccharomyces boulardii by spatiotemporal guidance for ameliorating inflammatory bowel disease

Abstract Background Compelling evidence indicates that dysregulated in the gut microbiota significantly contribute to the progression and pathogenesis of inflammatory bowel disease (IBD). Probiotic therapeutic interventions aimed at modulating the microbiota offer potential alternative approaches for treating IBD, but currently available probiotics often suffer from low intestinal colonization and limited targeting capability. Methods Here, we developed azido (N3)-modified Prussian blue nanozyme (PB@N3) spatio-temporal guidance enhances the targeted colonization of probiotics to alleviate intestinal inflammation. First, IVIS and Photoacoustic imaging demonstrated clickable PB@N3 targets intestinal inflammation, simultaneously, it scavenges reactive oxygen species (ROS). Subsequently, utilizing click chemistry “N3-DBCO” to spatio-temporally guide targeted colonization of dibenzocyclooctyne (DBCO)-modified Saccharomyces boulardii (S.Br@DBCO). The therapeutic effects of PB@N3 + S.Br@DBCO were verified by using the dextran sulfate sodium (DSS)-induced colitis mice model. Weight loss, fecal conditions, colon length and histopathological changes were monitored. Results PB@N3 targets and anchors to inflamed region. The "click" reaction between PB@N3 and S.Br@DBCO has excellent specificity and efficacy both in vivo and in vitro. Despite the complex physiological environment of IBD, "click" reaction can prolong the retention time of probiotics in the intestine. Dextran sulfate sodium (DSS)-induced colitis mice model, demonstrates that the combination of PB@N3 and S.Br@DBCO effectively enhances the colonization of probiotics, restores intestinal barrier function, and alleviates intestinal inflammation. Conclusion This is the first time that “click” chemistry was employed to combine fungal probiotics and nanozyme. PB@N3 spatio-temporal guidance enhances targeted colonization of S.Br@DBCO provides a promising medical treatment strategy for IBD.