Histopathological Changes Research Articles

NLRP3 inhibitor combined with Yimusake improves erectile dysfunction in rats with diabetes mellitus through the attenuation of pyroptosis

Erectile dysfunction (ED) is a prevalent complication associated with diabetes mellitus (DM), yet pharmacological treatments for diabetes-related erectile dysfunction (DMED) continue to be inadequate in clinical settings. Our previous studies have indicated that there is a close correlation between ED and pyroptosis, but the specific mechanism remains unclear. In this study, we sought to explore the therapeutic effects of DMED through the modulation of NLRP3, aiming to elucidate its potential molecular mechanisms. The DMED rat model was established via intraperitoneal injection of streptozotocin. The rats were randomly assigned to the control group, the DMED group, the Yimusake group, the MCC950 (NLRP3 inhibitor) group, and the MCC950+Yimusake group. Erectile function of rats was observed by measuring intracavernosal pressure (ICP) and mean arterial pressure (MAP). HE staining was performed to observe the histopathological changes in penile; immunofluorescence was performed to measure the level of CD31 (Platelet endothelial cell adhesion molecule-1) in penile. Besides, immunohistochemistry, RT-qPCR and Western blot were performed to demonstrate the expression of NLRP3, caspase-1, IL-1β and eNOS. After treatment with the MCC950 and Yimusake, the number of blood sinusoids and small vessels significantly reduced in penile tissue; NLRP3, caspase-1, IL-1β proteins and mRNA expression decreased, eNOS protein and mRNA expression increased. Compare with the Y group and the MCC950 group, MCC950+Yimusake group had a more significant effect. MCC950 and Yimusake might potentially suppress pyroptosis in the penile tissue of DMED rats by modulating the NLRP3/caspase-1 pathway, thus enhancing erectile function. This discovery could offer a promising therapeutic approach for individuals with DMED.

Xinglou Chengqi Decoction Protects against Cerebral Ischemia/Reperfusion Injury by Inhibiting Ferroptosis via SLC7A11/GPX4 Signaling.

Xinglou Chengqi decoction (XLCQD) is a Chinese formula that offers benefits in ischemic stroke. However, the underlying mechanism of the effects of XLCQD-mediated anti-ischemic stroke effects remains obscure. This study investigates the ferroptosis mechanism of XLCQD against cerebral ischemia/reperfusion (I/R) injury using rat models of middle cerebral artery occlusion/reperfusion (MCAO/R). Ferroptosis differs from traditional cell death pathways and is linked to oxidative stress-induced lipid peroxidation and glutathione (GSH) depletion, which is essential to the development of ischemic stroke. In this study, it is shown that XLCQD improves brain infarction, neurological dysfunction, and histopathological changes caused by MCAO/R exposure, and improving I/R-induced oxidative damage through inhibition of ferroptosis via (Solute Carrier Family 7 Member 11) SLC7A11/ (glutathione peroxidase 4) GPX4 pathway. Interestingly, it is found that XLCQD-mediated protection in I/Ris reversed by the silence of SLC7A11. XLCQD intervention significantly promotes GSH content and suppresses Reactive Oxygen Species(ROS), iron accumulation, as well as Malondialdehyde (MDA) generation, are markedly abrogated when SLC7A11 is knockdown by SLC7A11-shRNA transfection, indicating that SLC7A11 is the main target of XLCQD to further trigger intracellular events. In conclusion, XLCQD attenuates in vivo cerebral I/R injury by reducing ferroptosis via the SLC7A11/GPX4 pathway.

Acute Toxicity Assessment of Cream Ethanolic Extract of Vitex pinnata Bark from Central Kalimantan.

Vitex pinnata bark has the ability to scavenge free radicals and has antimicrobial activity, thus accelerating wound healing. The formulation of V. pinnata bark preparations has good organoleptic properties, homogenity, pH, dispersion, and absorption. We studied the acute toxicity of V. pinnata skin cream in Wistar rats according to OECD guidelines 402. Seven nulliparous, non-pregnant female Wistar rats were exposed 2000 mg/kg of body weight. There were no signs of toxicity in any behavioral, necropsy dan histopathological changes. The Globally Harmonized System classified V. pinnata cream extract as category 5 (unclassified) with LD50 more than two thousand mg/kg.

Uncovering cocoa shell as a safe bioactive food ingredient: Nutritional and toxicological breakthroughs

This work aimed to analyze the chemical composition of the cocoa shell flour (CSF) and aqueous extract (CSE) and conduct acute and sub-chronic toxicity assays to ensure safety. CSF was composed of 59.1 % dietary fiber, primarily insoluble fiber (48.1 %). CSE consisted entirely of soluble fiber (72.8 %), mainly pectic polysaccharides. The amino acid profile revealed CSF's potential as a high-quality plant protein source, whereas CSE was rich in free amino acids, especially leucine, valine, and alanine (59.6–62.5 mg 100 g−1). Higher levels of macroelements, especially potassium, were found in CSE. Silicon was the most abundant microelement, and heavy metal levels were within acceptable limits for both cocoa shell matrices. This study confirmed the bioactive potential of CSF and CSE, enriched in phenolic compounds and methylxanthines, including theobromine, caffeine, gallic acid, (+)-catechin, as well as N-phenylpropenoyl-l-amino acids. The high theobromine content in both matrices, especially in CSE (2605.3 mg 100 g−1), did not harm the mice's health; acute and sub-chronic toxicity studies demonstrated the safety of oral administration of CSF and CSE in mice, showing no lethality or remarkable histopathological changes. This research supports the safe use of CSF and CSE, establishing these ingredients as suitable for incorporation into food products.

Pseudoxanthoma elasticum: An unique entity in the coronary surgery.

Pseudoxanthoma elasticum is a rare connective tissue disease which affects the synthesis of the elastic fibres. It is an area of uncertainty for patients affected by this disease requiring surgical revascularization. We present a case report of a 43-year-old gentleman was known to have pseudoxanthoma elasticum syndrome (PXE) and coronary artery disease. He underwent successful off pump LIMA to LAD. Interestingly, this is the first case in UK and second one in the literature managed with an off-pump strategy. The predilection of the histopathological changes in PXE patients, should not prevent the routine use in of the left internal mammary artery as a standard conduit in CABG. Further studies are required to standardize the practice for surgical revascularization in the patients with PXE.

Anti-inflammatory effects of Tao Hong Si Wu Tang in mice with lung cancer and chronic obstructive pulmonary disease.

Lung cancer (LC) combined with chronic obstructive pulmonary disease (COPD) is a common combination of comorbidities. Anti-inflammation and modulation of oxidative/antioxidative imbalance may prevent COPD-induced LC, and are also crucial to the treatment of LC combined with COPD. Modern studies have shown that Tao Hong Si Wu Tang (THSW) has vasodilatory, anti-inflammatory, anti-fatigue, anti-shock, immunoregulatory, lipid-reducing, micronutrient-supplementing, and anti-allergy effects. To observe the effects of THSW on COPD and LC in mice. A total of 100 specific pathogen-free C57/BL6 mice were randomly divided into five groups: Blank control group (group A), model control group (group B), THSW group (group C), IL-6 group (group D), and THSW + IL-6 group (group E), with 20 mice in each group. A COPD mouse model was established using fumigation plus lipopolysaccharide intra-airway drip, and an LC model was replicated by in situ inoculation using the Lewis cell method. The blank control group exhibited a clear alveolar structure. The model control and IL-6 groups had thickened alveolar walls, with smaller alveolar lumens, interstitial edema, and several inflammatory infiltrating cells. Histopathological changes in the lungs of the THSW and THSW + IL-6 groups were less than those of the model control group. The serum IL-1β, IL-6, and TNF-α levels and IL-6R, JAK, p-JAK, STAT1/3, p-STAT1/3, FOXO, p-FOXO, and IL-7R expression levels in lung tissues of mice in the rest of the groups were significantly higher than those of the blank control group (P < 0.01). Compared with the model control group, the IL-6 group demonstrated significantly higher levels for the abovementioned proteins in the serum and lung tissues (P < 0.01), and the THSW group had significantly higher serum IL-1β, IL-6, and TNF-α levels and IL-7R expression levels in lung tissues (P < 0.01) but significantly decreased IL-6R, JAK, p-JAK, STAT1/3, p-STAT1/3, FOXO, p-FOXO, and IL-7R levels (P < 0.01). THSW reduces the serum IL-1β, IL-6, and TNF-α levels in the mouse model with anti-inflammatory effects. Its anti-inflammatory mechanism lies in inhibiting the overactivation of the JAK/STAT1/3 signaling pathway.

Protective Effect of Epigallocatechin-3-gallate against Hepatic Oxidative Stress Induced by tert-Butyl Hhydroperoxide in Yellow-Feathered Broilers.

Recent studies have shown that epigallocatechin-3-gallate (EGCG), as an effective antioxidant, could attenuate the oxidative damage, inflammation and necrosis in the liver in response to oxidative stress. The present study investigated whether oral administration of EGCG could effectively alleviate the hepatic histopathological changes and oxidative damage in yellow-feathered broilers induced by tert-butyl hydroperoxide (t-BHP). Broilers were exposed to 600 μmol t-BHP/kg body weight (BW) to induce oxidative stress by intraperitoneal injection every five days, followed by oral administration of different doses of EGCG (0, 20, 40 and 60 mg/kg BW) and 20 mg vitamin E (VE)/kg BW every day during 5-21 days of age. The results showed that t-BHP injection decreased (p < 0.05) body weight and the relative weight of the spleen; the enzyme activities of total antioxidant capacity (T-AOC), catalase (CAT) and total superoxide dismutase (SOD); and gene mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), CAT, SOD1, SOD2 and acetyl-CoA carboxylase (ACACA); as well as increased (p < 0.05) necrosis formation, malondialdehyde (MDA) content, reactive oxygen species (ROS)accumulation, and peroxisome proliferator activates receptor-α (PPARα) mRNA expression in the liver of yellow-feathered female broilers at 21 days of age. Treatment with 60 mg EGCG/kg BW orally could enhance antioxidant enzyme activities and reverse the hepatic damage induced by t-BHP injection by reducing the accumulation of ROS and MDA in the liver and activating the Nrf2 and PPARα pathways related to the induction of antioxidant gene expression (p < 0.05). In conclusion, intraperitoneal injection of t-BHP impaired body growth and induced hepatic ROS accumulation, which destroyed the antioxidant system and led to oxidative damage in the liver of yellow-feathered broilers from 5 to 21 days of age. It is suggested that EGCG may play an antioxidant role through the Nrf2 and PPARα signaling pathways to effectively protect against t-BHP-induced hepatic oxidative damage in broilers, and the appropriate dose was 60 mg EGCG/kg BW by oral administration.

Artemisinin Ameliorates the Neurotoxic Effect of 3-Nitropropionic Acid: A Possible Involvement of the ERK/BDNF/Nrf2/HO-1 Signaling Pathway.

Neurodegenerative disorders like Huntington's disease (HD) are a major threat to human health, with severe gait abnormalities and pathological changes (oxidative stress, neuroinflammation, and apoptosis) playing important roles in their development. The effects of artemisinin (ART) alone and in combination with the ERK antagonist PD98059 against 3-nitropropionic acid (3-NPA)-induced cell death and oxidative stress in SH-SY5Y cells were determined using the MTT and DCFH-DA assays, as well as RT-qPCR assays. In vivo, possible neuroprotective effects of ART (10, 20, and 40mg/kg i.p.) against the neurotoxicity generated by 21-day 3-NPA (10mg/kg i.p.) treatment was evaluated in rats by assessing behavioral parameters on days 1, 14, and 21. Further, various biochemical, inflammatory, apoptotic markers, histopathological changes, and protein expression were assessed using brain striatal samples. ART significantly mitigated the neurotoxic effect of 3-NPA in SH-SY5Y cells by regulating the mRNA expression of ERK, Bax, Bcl2, and cytochrome C. However, ART's neuroprotective activity was reduced in the presence of PD98059. Also, ART treatment for 21days substantially alleviated the behavioral impairments associated with 3-NPA toxicity. It reduced the oxidative stress induced by 3-NPA, as evidenced by the lower levels of MDA, nitrite, and improved catalase, SOD activity, and GSH levels. ART treatment restored 3-NPA-induced histopathological alterations in the striatal area. ART effectively suppressed neuroinflammatory (IL-6) and apoptotic markers (caspase 3 and 9), increasing BDNF levels and restoring the p-ERK1/2, Nrf2, and HO-1 expression. ART could exert its neuroprotective effect via antioxidant, anti-inflammatory, and antiapoptotic properties with a possible involvement of the ERK/BDNF/Nrf2/HO-1 pathway.

Enhancement of clinical signs in C3H/HeJ mice vaccinated with a highly immunogenic Leptospira methyl-accepting chemotaxis protein following challenge.

Leptospirosis is the most widespread zoonosis and a life-threatening disease in humans and animals. Licensed killed whole-cell vaccines are available for animals; however, they do not offer heterologous protection, do not induce long-term protection, or prevent renal colonization. In this study, we characterized an immunogenic Leptospira methyl-accepting chemotaxis protein (MCP) identified through a reverse vaccinology approach, predicted its structure, and tested the protective efficacy of a recombinant MCP fragment in the C3H/HeJ mice model. The predicted structure of the full-length MCP revealed an architecture typical for topology class I MCPs. A single dose of MCP vaccine elicited a significant IgG antibody response in immunized mice compared to controls (P < 0.0001), especially the IgG1 and IgG2a subclasses. The vaccination with MCP, despite eliciting a robust immune response, did not protect mice from disease and renal colonization. However, survival curves significantly differed between groups, and the MCP-vaccinated group developed clinical signs faster than the control group. There were differences in gross and histopathological changes between the MCP-vaccinated and control groups. The factors leading to enhanced disease process in vaccinated animals need further investigation. We speculate that anti-MCP antibodies may block the MCP signaling cascade and may limit chemotaxis, preventing Leptospira from reaching its destination, but facilitating its maintenance and replication in the blood stream. Such a phenomenon may exist in endemic areas where humans are highly exposed to Leptospira antigens, and the presence of antibodies might lead to disease enhancement. The role of this protein in Leptospira pathogenesis should be further evaluated to comprehend the lack of protection and potential exacerbation of the disease process. The absence of immune correlates of protection from Leptospira infection is still a major limitation of this field and efforts to gather this knowledge are needed.

Taraxasterol attenuates zearalenone-induced kidney damage in mice by modulating oxidative stress and endoplasmic reticulum stress

Taraxasterol is one of the bioactive ingredients from traditional Chinese herb Taraxacum, which exhibits multiple pharmacological activities and protective effects. However, the underlying influence and mechanism of its use against kidney damage caused from zearalenone (ZEA) remain unexplored. The ZEA-induced kidney damage model of mice was established by feeding diets containing ZEA (2 mg/kg), and taraxasterol (5 and 10 mg/kg) was administered by gavage for 28 days. Results demonstrated taraxasterol increased average daily gain (ADG) and average daily feed intake (ADFI), reduced feed-to-gain ratio (F/G) and kidney index of mice induced by ZEA. Taraxasterol alleviated histopathological changes of kidney, reduced ZEA residue and the levels of blood urea nitrogen (BUN), uric acid (UA), and creatinine (CRE). Concurrently, taraxasterol reduced the contents of oxidative stress indicator reactive oxygen species (ROS) and malondialdehyde (MDA), and increased the activities of antioxidant enzymes catalase (CAT), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px). Further, taraxasterol up-regulated the mRNA and protein expression of nuclear factor erythroid-2-related factor 2 (Nrf2), GSH-Px, NAD(P)H quinone oxidoreductase 1 (NQO1), and heme oxygenase-1 (HO-1), and down-regulated the mRNA and protein expression of KELCH like ECH associated protein (Keap1) in Nrf2/Keap1 pathway. Taraxasterol down-regulated the mRNA and protein expression of immunoglobulin binding protein (Bip), C/EBP homologous protein (CHOP), Bcl-2 associated X (Bax), cysteine protease (Caspase)-12, and Caspase-3, and up-regulated B-cell lymphoma 2 (Bcl-2) expression in endoplasmic reticulum stress pathway. This study suggests that taraxasterol attenuates ZEA-induced mouse kidney damage through the modulation of Nrf2/Keapl pathway to play antioxidant role and endoplasmic reticulum stress pathway to enhance anti-apoptotic ability. It will provide a basis for taraxasterol as a potential drug to prevent and treat ZEA-induced kidney damage.

A Cross-Sectional Study Comparing Placental Characteristics in Pregnancy-Induced Hypertension and Sickle Cell Anaemia.

Placental health plays a critical role in pregnancy outcomes as it serves as the interface between the mother and fetus. High-risk pregnancies, such as those complicated by pregnancy-induced hypertension (PIH) and sickle cell anaemia (SCA), are associated with significant alterations in placental morphology and histopathology, potentially leading to adverse maternal and fetal outcomes. While previous studies have explored placental changes associated with either PIH or SCA, nobody has comparatively analysed these conditions to understand their unique and overlapping effects on placental pathology. This study aimed to compare and contrast the morphological and histopathological changes in the placenta associated with high-risk pregnancies likePIH and SCA. A comparative analysis was conducted using data fromstudies at two different tertiary care centres. The study populations included 100 pregnant women diagnosed with PIH and 56 pregnant women with SCA, alongside a matched control groupof 100 healthy pregnant women for the PIH group and 56 healthy pregnant women for the SCA group. Inclusion criteria were restricted to singleton pregnancies in women aged 18 to 35 years with gestational ages ranging from 28 to 40 weeks. Following delivery, placental specimens were collected, and various parameters such as weight, volume, surface area, number of cotyledons, and umbilical cord attachment were meticulously measured. Histopathological examinations were conducted to identify specific pathological features like infarcts, calcifications, syncytial knots, and fibrin deposition. Data analysis was performed using SPSS software version 25.0 (IBM Corp., Armonk, NY, USA), with a p-value of less than 0.05 considered statistically significant. The study revealed significant differences in placental parameters between the PIH and SCA groups compared to their respective controls. Placental weight, volume, and surface area were significantly reduced in both the PIH and SCA groups, with more pronounced reductions observed in PIH (p < 0.001). The umbilical cord attachment was predominantly marginal in the PIH group (75%), compared to a central attachment in the SCA group (70%), suggesting different patterns of placental development. Histopathological analysis demonstrated a higher incidence of infarcts (65% vs. 30%), calcifications (80% vs. 45%), syncytial knots (90% vs. 50%), and fibrin deposition (70% vs. 35%) in the PIH group compared to the SCA group, indicating more severe placental pathology in PIH. This study offered a comprehensive comparison of placental changes in pregnancies affected by PIH and SCA. It identified both structural abnormalities in terms of placental weight, volume, and surface area and distinctive pathological features like infarctions, calcifications, syncytial knot formation, and fibrin deposits in the placenta. When compared to control groups, these findings strongly suggest that placental dysfunction is a key contributor to the adverse outcomes associated with these high-risk pregnancies.

Deciphering the ameliorative effect of Aloe vera (L.) burm. F. extract on histopathological alterations in Streptozotocin-induced WNIN/GR-ob rats

Ethnopharmacological relevanceNatural products have emerged as a novel source in the management of non-communicable diseases, more so in diabetes mellitus and its comorbidities. Aloe vera is widely recognized for its anti-hyperglycemic and anti-hyperlipidemic properties and numerous researchers have identified component (s) from Aloe vera attributing to these therapeutic effects. Aim of the studyThe current work was undertaken to gain insight into the protective effect of Aloe vera (L.) Burm. f. extract to study the cytoarchitecture/histopathological alterations in the target organs in mutant Obese WNIN/GR-Ob rats that were made frank diabetic with streptozotocin. Materials and methodsRats were divided into five groups. 1)WNIN-GR-Ob/control group 2)WNIN-GR-Ob treated with STZ 3)WNIN-GR-Ob + STZ + Sitagliptin 4)WNIN-GR-Ob + STZ + Aloe vera 5)WNIN-GR-Ob/control group + Aloe vera. Histopathological analysis of the pancreas, kidney, liver, and adipocytes was done after 4 weeks of treatment. ResultsThe histopathological examination of STZ-induced diabetic rats revealed significant changes in all the vital organs including cell infiltration, degeneration, and necrosis. Treatment with A. vera negated most of the histopathological changes seen in STZ induced rats. Sitagliptin—which served as a positive control in the present study—reversed the alterations seen in streptozotocin rats. ConclusionConsidering the hypoglycaemic and hypolipidemic activities of Aloe vera that have been previously demonstrated by us, the present study further re-instates the therapeutic efficacy of Aloe vera towards vital organs. It was able to restore islet cells and reduce β-cell damage. In addition, it was also able to aid in kidney tubular regeneration and reverse the degenerative changes brought on by streptozotocin on liver. Further, Aloe vera treated group exhibited moderate hyperplasia with decreased size of adipocytes and reduced macrophage infiltration. Thus, our findings advocate its application as an important nutraceutical in the therapeutic management of diabetes mellitus and associated complications.

Unveiling the Nutritional Profile and Safety of Coffee Pulp as a First Step in Its Valorization Strategy.

The coffee pulp, a significant by-product of coffee processing, is often discarded but has potential for recycling and high-value uses. This study aimed to investigate the chemical composition of two coffee pulp ingredients, a flour (CPF) and an aqueous extract (CPE), and conducted acute and sub-chronic toxicity assays to determine their safety. The proximate composition revealed the high fiber content of both ingredients; the CPF mainly contained insoluble fiber, while CPE consisted exclusively of soluble pectic polysaccharides. The CPF had higher concentrations of amino acids and a better balance of essential/non-essential amino acids, whereas the CPE exhibited higher concentrations of free amino acids, ensuring higher bioavailability. Both ingredients showed elevated mineral content, while heavy-metal concentrations remained within acceptable limits. This study established the bioactive potential of the CPF and the CPE, demonstrating the high content of caffeine and gallic, protocatechuic, and 4-caffeoylquinic acids. The toxicity studies revealed that the CPF and the CPE exhibited safety when orally administered to mice. Administered doses were non-toxic, as they did not induce lethality or adverse effects in the mice or produce significant histopathological or biochemical adverse changes. This study represents a first step in valorizing the CPF and the CPE as safe novel food ingredients with health benefits for functional and nutritional foods.

Distribution of mechanical properties of native human ligamentum flavum depending on histopathological changes

This study aimed to characterize the mechanical properties of native human ligamentum flavum (LF) and correlate them with histopathological changes. Mechanical property gradients across the cranial, medial, and caudal regions of LF were mapped and compared with histological sections. We also compared lumbar spinal stenosis (LSS) samples with disc herniation (DH) samples as reference material to identify differences in mechanical properties and histopathological features. Our results revealed significant heterogeneity in LF mechanical properties, with local variations correlating with specific histopathological changes such as chondroid metaplasia and loss of elastic fibers. These findings underscore the importance of considering LF heterogeneity in mechanical characterization and provide insights into its behavior under pathological conditions.

Potential Protective Effect of Hesperidin (Vitamin P) against Glyphosate-Induced Spermatogenesis Damage in Male Rats: Biochemical and Histopathological Findings on Reproductive Parameters.

The aim of this study was to investigate the effectiveness of hesperidin (HES) on testicular histopathological changes, biochemical changes, and semen characteristics in rats exposed to glyphosate (GLP). The control group was given a normal diet devoid of GLP and HES, the HES group was given 100 mg/kg/day HES with the normal diet, the GLP group was given GLP at the LD50/10 dose of normal feed, which was 787.85 mg/kg/day, and the GLP + HES group was given normal feed containing 787.85 mg/kg/day LD50/10 dose of GLP in addition to 100 mg/kg/day HES. GLP administration reduced sperm motility, sperm plasma membrane integrity, glutathione levels, and total antioxidant levels in the testicular tissues of rats. Moreover, it caused an increase in right testis and left epididymis weights, abnormal sperm counts, malondialdehyde levels, total oxidant status, and DNA damage. The HES treatment showed curative effects on these parameters. Furthermore, HES was effective in lessening the histopathological damage that was caused by GLP. The results showedthat HES protects spermatological parameters and DNA integrity, improves antioxidant defenses, and lowers the damage and lipid peroxidation caused by GLP in testicular tissue.

Pterocarya hupehensis Skan total flavones ameliorate rheumatoid arthritis in rats by suppressing formation of neutrophil extracellular traps

To investigate the therapeutic mechanism of Pterocarya hupehensis Skan total flavonoids (PHSTF) for rheumatoid arthritis (RA). Twenty-five male SD rats were randomly divided into normal control group, RA model group, PHSTF treatment (45 and 90 mg/kg) groups, and Tripterygium glycosides (TPG) tablet (10 mg/kg) group (n=5). Except for those in the normal control group, all the rats were subjected to collagen-induced arthritis (CIA) modeling using a secondary immunization method, after which PHSTF and TPG were administered via gavage once daily for 4 weeks. After the treatments, serum levels of TNF-α and IL-1β were measured using ELISA, and ankle joint pathologies were assessed with HE staining; the expression of citrullinated histone H3 (Cit-H3), a neutrophil extracellular trap (NET) marker, in the ankle joints was evaluated with immunohistochemistry. In primary cultures of rat peripheral blood neutrophils stimulated with phorbol ester (PMA), the effects of PHSTF (100 and 200 μg/mL) on the expressions of Cit-H3, peptidylarginine deiminase 4 (PADI4), neutrophil elastase (NE), and myeloperoxidase (MPO) were examined with Western blotting; immunofluorescence assay was used to observe Cit-H3 expression and NET formation in the cells. In the CIA rat models, PHSTF significantly alleviated ankle swelling, decreased serum levels of TNF-α and IL-1β, improved histopathological changes in the ankle joints, and reduced Cit-H3 expression in both the serum and ankle joint cartilage. In the isolated rat neutrophils, PHSTF showed no significant effect on cell viability but strongly inhibited PMA-induced NET release. PHSTF can alleviate RA by inhibiting the formation of NETs.

Synthetic bacterial consortia transplantation attenuates vaginal inflammation and modulates the immune response in a mouse model of Gardnerella vaginalis-induced bacterial vaginosis

This study aimed to evaluate the efficacy of synthetic bacterial consortia transplantation (SBCT) and compare it with VMT (vaginal microbiota transplantation) in a mouse model of Gardnerella vaginalis-induced Bacterial vaginosis (BV). A murine model of G. vaginalis-induced BV was established, and mice were treated with SBCT, VMT, or saline. Histopathological changes, inflammatory cytokine levels, pro-inflammatory biomarker expression, helper T cell transcription factor expression, and vaginal microbiota composition were assessed. SBCT and VMT effectively suppressed G. vaginalis growth, reduced inflammation, and restored vaginal microbiota diversity. Both treatments attenuated epithelial damage, downregulated pro-inflammatory cytokines (IL-1β and IL-8), and upregulated the anti-inflammatory cytokine IL-10. SBCT and VMT also inhibited NF-κB activation, suppressed IL-17 expression, and enhanced Foxp3 expression in vaginal tissues. SBCT is a promising therapeutic approach for treating BV, as it effectively modulates the immune response and restores vaginal microbiota diversity in a mouse model of G. vaginalis-induced BV.

Integrated transcriptomics and proteomics analyses reveal the ameliorative effect of hepatic damage in tilapia caused by polystyrene microplastics with chlorella addition

Fish exhibit varying responses to polystyrene microplastics (MPs) depending on particle size. Previous studies suggested that microorganisms adhering to the surface of MPs can induce toxic effects. In this study, Tilapia were exposed to MPs of control (group A), 75 nm (B), 7.5 μm (C), 750 μm (D), as well as combinations of all sizes (E) and 75 nm MPs with Chlorella vulgaris addition (F) for 7, 10 and 14 days. Histopathological changes in liver of tilapia were assessed using enzyme activities, transcriptomics and proteomics. The results showed that in groups combined MPs of different particle sizes and those supplemented with chlorella, MPs were localized on the surface of goblet cells, leading to vacuoles, constricted hepatic sinuses and nuclei displacement. Exposure to 7.5 and 750 μm MPs significantly increased the contents of fatty acid synthase (FAS), adenosine triphosphate (ATP), acetyl-CoA carboxylase (ACC), lipoprotein lipase (LPL), total cholesterol (TC), total triglyceride (TG) contents at 7 and 10 days. In particular, cytochrome p450 1a1 (EROD), reactive oxygen species (ROS) and superoxide dismutase (SOD) were markedly elevated following exposure to MPs. Apoptotic markers caspase-3, and inflammatory markers, including tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β), had a similar upward trend in comparisons of group C vs A at 7 d, group D vs A at 14 d. The peroxisome proliferator activated receptor (PPAR) signaling pathway, spliceosome, was highly enriched during the 7-day exposure of medium sized MPs, while largest MPs in the comparison of group D vs A at 14 d activated pathways such as phagosome, apoptosis, salmonella infection. Transcriptomic analysis revealed that after 14 days, the kyoto encyclopedia of genes and genomes (KEGG) pathways associated with protein processing in endoplasmic reticulum and the PPAR signaling has been significantly enriched in the Chlorella-supplemented group, which was further confirmed via the proteomic analysis. Overall, the findings highlight the size-dependent effects of MPs on histopathological changes, gene and protein expression in the liver of tilapia, and C. vulgaris effectively attenuated liver damages, likely through modulation of endoplasmic reticulum protein processing and PPAR signaling pathways.

Seasonal fluctuations of heavy metal accumulation and gastrointestinal helminth induce oxidative stress and histological lesions in resident catfish Clarias gariepinus

The exposure of the African catfish (Clarias gariepinus) to various environmental contaminants leads to physiological and histological alterations. Therefore, the study aimed to assess the impact of seasonal variations of ecological contaminants as external stressors and internal stressors via helminth infections on oxidative stress and histopathology in resident C. gariepinus at EL-Salam Canal, Egypt. Seasonal water and sediment samples were collected to assess physicochemical parameters and heavy metals. The length, weight, and sex were recorded for each fish sample. The gastrointestinal tract was dissected from the visceral cavity, and the helminths were extracted and identified using SEM photos based on their morphological characteristics. The parasitic dominance in the gastrointestinal helminths was calculated. The heavy metal concentrations, bioaccumulation (BAF), and biosedimentation (BSF) factors were considered in the muscles and intestine. Specimens of muscles and intestines were removed to determine oxidative status. In addition, pieces of skin, muscles, stomachs, and intestines were subjected to light microscopy to determine the alterations. The calcium, magnesium, and potassium levels were within safe limits. Sulfate levels consistently remained below the maximum permissible thresholds throughout the seasons. Among the heavy metals examined, the highest accumulation was found in the intestinal tissues of C. gariepinus, while muscle tissues showed lower levels. The variability in metal concentrations across water, sediment, muscles, and intestines underscores the different capacities of these environments to accumulate heavy metals. The elevated metal levels in fish tissues raise concerns about potential health risks for humans who consume contaminated fish, highlighting significant bioaccumulation within the food chain. The result indicated that Cu in the sediment samples was associated with parasite abundance. The dual stress from environmental pollutants and parasitic infections exacerbates oxidative stress and causes notable histopathological changes in the tissues of the catfish. These results highlight the intricate interplay between external and internal stressors, emphasizing the need for continuous monitoring and management of aquatic ecosystems to safeguard the health of resident fish populations. It provides insight into how these factors affect fish health.

Fluxapyroxad induced toxicity of earthworms: Insights from multi-level experiments and molecular simulation studies

Fluxapyroxad, an emerging succinate dehydrogenase inhibitor fungicide, is widely used due to its excellent properties. Given its persistence in soil with a 50 % disappearance time of 183–1000 days, it is crucial to evaluate the long-term effects of low-dose fluxapyroxad on non-target soil organisms such as earthworms (Eisenia fetida). The present study investigated the impacts of fluxapyroxad (0.01, 0.1, and 1 mg kg−1) on Eisenia fetida over 56 days, focusing on oxidative stress, digestive and nervous system functions, and histopathological changes. We also explored the mechanisms of fluxapyroxad-enzyme interactions through molecular docking and dynamics simulations. Results demonstrated a significant dose-response relationship in the integrated biomarker response of 12 biochemical indices. Fluxapyroxad altered expression levels of functional genes and induced histopathological damage in earthworm epidermis and intestines. Molecular simulations revealed that fluxapyroxad is directly bound to active sites of critical enzymes, potentially disrupting their structure and function. Even at low doses, long-term fluxapyroxad exposure significantly impacted earthworm physiology, with effects becoming more pronounced over time. Our findings provide crucial insights into the chronic toxicity of fluxapyroxad and emphasize the importance of long-term, low-dose studies in pesticide risk assessment in soil. This research offers valuable guidance for the responsible management and application of fungicides.