Histopathological Observations Research Articles

Role of Aluminium in Alzheimer’s disease: Ultrastructural Study in Rat Hippocampus

Background: Exposure to high levels of aluminium (Al) leads to neurotoxicity. Hippocampus is one of the preferred sites of aluminium accumulation. Nevertheless, the role of Al in Alzheimer’s disease (AD) remains controversial and there is little proof directly interlinking Al to AD. Aims: The present study was undertaken to find out the occurrence of AD pathogenesis in Hippocampus under moderate aluminium exposure in rats. Materials and Methods: Adult rats were divided into control (C) and aluminium treated (E) groups having eight animal each. The rats in group E were exposed to aluminium 4.2 mg/kg body weight for three months with due approval from Institute Animal Ethics Committee. The hippocampus was processed for histopathological and electron microscopy observation. Results: Moderate Al intake produces significant reduction in the count of Pyramidal cells in hippocampus identified by shrunken cells as well as pyknosis in cell bodies. The differences between the cell numbers in all groups were found to be statistically significant (P  0.05). Cornu Ammonis (CA) exhibited significantly reduced nissl bodies with a marked reduction in neuronal cell loss. Neurofibrillary tangle and plaques were not seen in the given dose of Al exposure. Electron microscopy from experimental group showed that the majority of neurons were disintegrating, the nuclear membrane has ruptured, and nucleoli appeared significantly distorted. The chromatin condensed and the mitochondria had disintegrated. Many vacuoles and lipofuscin sediment in cytoplasm, as compared to the control group noted. Conclusion: Present data demonstrated that moderate chronic aluminium exposure 4.2mg/kg body weight induced neurodegeneration in hippocampus but not significant for Alzheimer’s disease pathogenesis. KEYWORDS: Aluminium, Hippocampus, Alzheimer’s Disease, Neurodegeneration, Histopathology, Electron microscopy.

Pathological observation and transcriptomic analysis of thymus injury in PRRSV-infected piglets

The thymus, the central immune organ in mammals, plays an important role in immune defense. Porcine reproductive and respiratory syndrome virus (PRRSV) infection in piglets can cause thymus injury and immunosuppression. However, the mechanisms of thymus injury remain unknown. This study was aimed at investigating the specific manifestations of thymus injury through the construction of a PRRSV-infected piglet model and histopathological observation. In this study, fourteen 40-day-old PRRSV-free piglets were randomly divided into two groups, eleven of which were intramuscularly injected with 3 mL of PRRSV WUH3 virus suspension (106 PFU /mL) in the infection group, and three of which were sham-inoculated with 3 mL of RPMI-1640 medium in the control group. Clinical necropsy and samples collection were performed on day 8 after artificial infection. With the Illumina platform, the transcriptomes of piglet thymus tissues from infected and control piglets were sequenced to explore the relationships of differentially expressed genes (DEGs) and signaling pathways with thymus injury. The immune organs of PRRSV-infected piglets were severely damaged. The histopathological findings in the thymus indicated that PRRSV infection was associated with a large decrease in lymphocytes, cell necrosis and cell apoptosis; an increase in blood vessels and macrophages; thymic corpuscle hyperplasia; and interstitial widening of the thymic lobules. The transcriptomic analysis results revealed that the Gene Ontology functions of DEGs were enriched primarily in biological processes such as angiogenesis, regulation of angiogenesis and positive regulation of cell migration. Moreover, greater numbers of blood vessels and macrophages were observed in the thymus in PRRSV-infected than control piglets. KEGG pathway enrichment analysis revealed that the DEGs were significantly enriched in the Toll-like receptor signaling pathway, chemokine signaling pathway, IL-17 signaling pathway and TNF signaling pathway. The expression of TLR8, IRF5, the chemokines CCL2, CCL3L1 and CCL5; and their receptors CCR1, CCR2 and CCR5 was significantly up-regulated in PRRSV infection, thus suggesting that these cytokines were associated with the pathological processes of thymus injury.

Anti-TLR4 IgG2 Prevents Acetaminophen-induced Acute Liver Injury through the Toll-like Receptor 4/MAPKs Signaling Pathway in Mice.

Acetaminophen (APAP) is a widely used antipyretic and analgesic. If taken in excess, it can cause severe drug-induced acute liver injury. The purpose of this study was to investigate the effects of anti-TLR4 IgG2 on APAP-induced liver injury and its underlying mechanisms. We injected APAP into the abdominal cavity of mice to establish a liver injury model. Mice were divided into the control group, APAP group, and APAP + anti-TLR4 IgG2 group. In order to verify the implication of the toll-like receptor4 and mitogen-activated protein kinases activation (TLR4/MAPKs) signaling pathway, mice were intraperitoneally injected with a TLR4 / MAPKs inhibitor anti-TLR4 IgG2. We evaluated the effects of TLR4 IgG2 on the antioxidant, anti-apoptotic, anti-inflammatory, and liver histopathology of APAP mice. In addition, the expression of the TLR4 / MAPKs signaling pathway was detected by Western blot. Our study showed that APAP mouse models were successfully established; however, pretreatment with anti-TLR4 IgG2 alleviated APAP-induced hepatic injury, as evidenced by the 24-h survival rate. Meanwhile, anti-TLR4 IgG2 prevented the elevation of serum biochemical parameters and lipid profile. Furthermore, compared with the APAP group, hepatic antioxidants, including 3- Nitrotyrosine, high mobility group protein B1, superoxide dismutase, catalase, and glutathione, were increased in APAP + anti-TLR4 IgG2 group. In contrast, a significant decrease was observed in the levels of the malondialdehyde, which is a lipid peroxidation product. Moreover, the western blotting analysis showed that anti-TLR4 IgG2 treatment inhibited the activation of the apoptotic pathway by increasing Bcl-2 and decreasing Bax, P53, and cleaving caspase-3 / caspase-3 protein expression. These results were further validated by TUNEL staining and immunohistochemical. Histopathological observation also revealed that pretreat-ment with anti-TLR4 IgG2 could significantly reverse hepatocyte inflammatory infiltration, congestion, and necrosis in liver tissues by APAP. Importantly, anti-TLR4 IgG2 effectively alleviated APAP-induced liver injury by inhibiting tolllike receptor4 and mitogen-activated protein kinases activation signaling pathways (TLR4/MAPKs). The results clearly suggest that the underlying molecular mechanisms in the hepatoprotection of anti-TLR4 IgG2 in APAP-induced hepatotoxicity may be due to its antioxidation, anti-apoptosis, and anti-inflammation effects through inhibition of the TLR4/MAPKs signaling axis.

Long Non-Coding RNA Plasmacytomavariant Translocation1 as ceRNA Adsorbs miR-195 to Relieve Osteoarthritis Symptoms

This study aimed to investigate the role of the long non-coding RNA plasmacytomavariant translocation1 (LncRNA PVT1) in the development of osteoarthritis (OA). The study used mice and performed DMM surgery to establish an OA model. PVT1 and miR-195 agomir were inhibited in the knee joints, and cartilage tissue specimens were collected for gene expression analysis, apoptotic protein detection, histopathological observation, and Mankin’s score evaluation. Enzymelinked immunosorbent assay (ELISA) was used to measure the levels of inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in peripheral blood and cartilage tissues. Terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining was used to detect cartilage cell apoptosis. The results indicated that the OA group had higher plasmacytomavariant translocation1 (PVT1) expression and lower miR-195 expression than the sham-operated group. PVT1 was found to act as a molecular sponge for miR-195 through a dual-luciferase reporter assay. Knocking down PVT1 or upregulating miR-195 significantly attenuated inflammation and apoptosis in the bone joints of OA mice. This study provides new insights into the mechanism of OA progression and suggests that regulating the PVT1-miR-195 axis may be beneficial for improving the inflammatory microenvironment of cartilage tissue and apoptosis.

Esculin alleviates LPS-induced acute lung injury via inhibiting neutrophil recruitment and migration.

Acute lung injury (ALI) poses a serious threat to human health globally, particularly with the Coronavirus 2019 (COVID-19) pandemic. Excessive recruitment and infiltration of neutrophils is the major etiopathogenesis of ALI. Esculin, also known as 6,7-dihydroxycoumarin, is a remarkable compound derived from traditional Chinese medicine Cortex fraxini. Accumulated evidence indicates that esculin has potent anti-inflammatory effects, but its pharmaceutical effect against ALI and potential mechanisms are still unclear. This study evaluated the protective effect of esculin against ALI by histopathological observation and biochemical analysis of lung tissues and bronchoalveolar lavage fluid (BALF) in lipopolysaccharide (LPS)-challenged ALI mice in vivo. The effects of esculin on N-formyl-met-leu-phe (fMLP)-induced neutrophil migration and chemotaxis were quantitatively assessed using a Transwell assay and an automated cell imaging system equipped with a Zigmond chamber, respectively. The drug affinity responsive target stability (DARTS) assay, in vitro protein binding assay and molecular docking were performed to identify the potential therapeutic target of esculin and the potential binding sites and pattern. Esculin significantly attenuated LPS-induced lung pathological injury, reduced the levels of pro-inflammatory cytokines in both BALF and lung, and suppressed the activation of NF-κB signaling. Esculin also significantly reduced the number of total cells and neutrophils as well as myeloperoxidase (MPO) activity in the BALF. Esculin impaired neutrophil migration and chemotaxis as evidenced by the reduced migration distance and velocity. Furthermore, esculin remarkably inhibited Vav1 phosphorylation, suppressed Rac1 activation and the PAK1/LIMK1/cofilin signaling axis. Mechanistically, esculin could interact with β2 integrin and then diminish its ligand affinity with intercellular adhesion molecule-1 (ICAM-1). Esculin inhibits β2 integrin-dependent neutrophil migration and chemotaxis, blocks the cytoskeletal remodeling process required for neutrophil recruitment, thereby contributing to its protective effect against ALI. This study demonstrates the new therapeutic potential of esculin as a novel lead compound.

The Healing Effect of Biodegradable Scaffolds Treated with Bone-Marrow Obtained Mesenchymal Stem Cells on Major Tendon Damage in the Dog as a Model.

The present study aimed to evaluate the implantation of decellularized small intestinal submucosa- extracellular matrix )SIS-ECM( seeded with bone marrow mesenchymal stem cells (BM-MSCs) to repair full-thickness Achilles tendon defect. For this purpose, 20 healthy adult stray dogs aged 8-12 months old (15±3 kg of weight) were enrolled in this study under an aseptic environment and general anesthesia. A 1.5 cm-long segment-based resection was performed in the mid-substance of the Achilles tendon in the control group (n=10) that did not receive treatment. While, in the experimental group (n=10), regarding the defect of the tendon, the stumps were bridged with decellularized SIS seeded with BM-MSCs (5×106) cells implanted. Afterward, the stumps of the tendon were sutured using the modified Kessler technique (4-0) polypropylene thread. The biomechanical observations of the tendon defect showed an increase in the tensile strength in the experimental group, compared to the control animals. It should be mentioned that this difference was significant (P≤0.05). Histopathological observations of biopsies harvested after the 4th, 8th, and 12th weeks revealed that the implanted graft had seeded with MSCs enhanced high-quality cellular infiltration and the host tissue healing was improved. Similar to the normal tendon, a dense organized collagenous tissue with high cellularity and vascularity was observed due to the presence of the remodeled ECM. However, the arrangement of collagen-fiber-derived connective tissue appeared to be more dominant than that in the experimental group, with less adhesion in the 12th week post-treatment. These findings suggest that the BM-MSCs inoculated with SIS can be employed to repair a damaged Achilles tendon due to the fact that this combination enhances the regeneration of the affected tendon.

The Effectiveness of Ethanol Extract of Ketapang Leaves (Terminalia catappa L.) as a Larvicidal Against Aedes aegypti Mosquito Larvae

This study aimed to determine the effectiveness of the ethanol extract of ketapang leaves (Terminalia catappa) on mortality and histopathology of Ae. aegypti instar III as an effort to control Ae. aegypti as a disease vector without causing resistance to the larvae. The extraction process uses the maceration method. This type of research is a laboratory experiment with research subjects Ae. aegypti instar III of 700 individuals, which were divided into 7 treatment groups (1 negative control group using distilled water and 1% DMSO, 1 positive control group using 1000 ppm abate, ethanol extract of ketapang leaves with concentrations of 2000 ppm, 2500 ppm, 3500 ppm, 5000 ppm, 7000 ppm) and 4 repetitions. The results of this study indicate that the negative control (K-) mortality rate is 0% and the positive control (K+) mortality percentage is 100%. The results of the ethanol extract of ketapang leaves (Terminalia catappa) on the mortality of Ae. aegypti instar III in treatment one (P1) the percentage of mortality was 17%, treatment two (P2) the percentage of mortality was 21%, treatment three (P3) the percentage of mortality was 23%, treatment four (P4) the percentage of mortality was 27%, and treatment five (P5) the percentage of mortality is 42%. Histopathological observations on larvae that died after being exposed to ethanol extract of ketapang leaves (Terminalia catappa) for 24 hours damaged the basement membrane, peritropic membrane and epithelial cells. It can be concluded that the ethanol extract of ketapang leaves (Terminalia catappa) is effective as a larvicidal against Ae. aegypti instar III but can cause damage to the midgut and cause death of the larvae.

Fitness assessment of Mytilus galloprovincialis Lamarck, 1819 after exposure to herbicide metabolite propachlor ESA

The lack of data on the chronic effects of chloroacetanilide herbicide metabolites on non-target aquatic organisms creates a gap in knowledge about the comprehensive impacts of excessive and repeated pesticide use. Therefore, this study evaluates the long-term effects of propachlor ethanolic sulfonic acid (PROP-ESA) after 10 (T1) and 20 (T2) days at the environmental level of 3.5 μg.L−1 (E1) and its 10x fold multiply 35 μg.L−1 (E2) on a model organism Mytilus galloprovincialis. To this end, the effects of PROP-ESA usually showed a time- and dose-dependent trend, especially in its amount in soft mussel tissue. The bioconcentration factor increased from T1 to T2 in both exposure groups – from 2.12 to 5.30 in E1 and 2.32 to 5.48 in E2. Biochemical haemolymph profile and haemocyte viability were not affected by PROP-ESA exposure. In addition, the viability of digestive gland (DG) cells decreased only in E2 compared to control and E1 after T1. Moreover, malondialdehyde levels increased in E2 after T1 in gills, and DG, superoxidase dismutase activity and oxidatively modified proteins were not affected by PROP-ESA. Histopathological observation showed several damages to gills (e.g., increased vacuolation, over-production of mucus, loss of cilia) and DG (e.g., growing haemocyte trend infiltrations, alterations of tubules). This study revealed a potential risk of chloroacetanilide herbicide, propachlor, via its primary metabolite in the Bivalve bioindicator species M. galloprovincialis. Furthermore, considering the possibility of the biomagnification effect, the most prominent threat poses the ability of PROP-ESA to be accumulated in edible mussel tissues. Therefore, future research about the toxicity of pesticide metabolites alone or their mixtures is needed to gain comprehensive results about their impacts on living non-target organisms.

A strontium/vancomycin composite coating on titanium implants for preventing bacterial infection and improving osseointegration

Postoperative bacterial infection and poor osseointegration with surrounding bone tissue are not only the critical causes of implant failure but also the major challenges in clinical application. Hereby, Titania (TiO2) microporous/nanofiber coating was firstly prepared on the surface of titanium (Ti) material by alkali-heat treatment process, and strontium (Sr)-doped coating (TS) was then prepared by hydrothermal synthesis. Finally, polydopamine-assisted co-deposition technology was utilized to deposit vancomycin (Van) onto TS surface (TSV), thus obtaining a medical composite coating with antibacterial property and improved osseointegration ability. According to antibacterial test, TSV has not only a 99.99% killing effect on methicillin-resistant Staphylococcus aureus (MRSA), but also a good anti-biofilm ability. The cell experiments showed that TSV could enhance the adhesion and proliferation of osteoblasts. Compared with pure Ti and TS, TSV induces the formation of bone apatite in simulated body fluid (SBF) test quickly, indicating that the coating has good biological activity. According to histopathological observation and bone morphology analysis, TSV showed highly effective infection prevention and excellent osteointegration in vivo. These test results suggested that it is feasible to prevent implant-associated infection and promote osseointegration by functionalization of the implant, so as to provide a fresh direction for clinical research.

Effect of Phyllosilicates As Toxin Binder On Productivity, Intestinal Morphology, and Liver Toxicity in Broiler Fed Afb1 Contaminated Feed

Aflatoxin B1 is a toxin produced by the fungus Aspergillus flavus which reduces the development and function of organs in broilers. The aim of this study was to determine the effect of adding binder toxin from different bentonite to feed contaminated with AFB1 on productivity, intestinal morphology, and liver toxicity in broilers. A total of 60-day old chick male broilers were placed in 12 pens. Each treatment consisted of three replicates, each replicate containing five broilers. Treatment in the study consisted of P0 (control, basal diet, without the addition of AFB1), P1 (P0 + 100 µg/kg AFB1 + 4 g/kg calcium bentonite Type A), P2 (P0 + 100 µg/kg AFB1 + 4 g/kg calcium bentonite Type B), and P3 (P0 + 100 µg/kg AFB1 + 4 g/kg calcium bentonite Type B + kerolite + saponite). Treatment diets were given to broilers from day 22 to 35 (finisher phase). The results showed that the toxin binder on AFB1 contaminated feed had no effect on feed consumption, body weight gain and feed conversion (p>0.05). Addition of toxin binder on AFB1 contaminated feed increased the relative weight of the duodenum (p=0.024), although P3 was not significantly different. Treatments had no effect on villus length, crypt depth, and ratio of villus length to crypt depth (p>0.05), but decreased villus width (p=0.013). The addition of toxin binder tended to decrease the villus area (p=0.055). SGOT and SGPT did not show differences between treatments. AFB1 contamination with the addition of toxin binder showed signs of toxicity on liver histopathological observations. Based on the research, it can be concluded that the addition of binder toxin in feed contaminated with AFB1 can reduce the negative effect on the development of intestinal villus and chemical effect to the liver. Toxin binder Type B has the best efficacy for reduce the negative effect

Geniposide ameliorates dextran sulfate sodium-induced ulcerative colitis via KEAP1-Nrf2 signaling pathway

Ethnopharmacological relevanceThe dried fruit of Gardenia jasminoides Ellis (Zhizi in Chinese) is a traditional medicine used for thousands of years in China, Japan and Korea. Zhizi was recorded in Shennong Herbal, as a folk medicine, it reduces fever and treats gastrointestinal disturbance with antiphlogistic effects. Geniposide, an iridoid glycoside, is an important bioactive compound derived from Zhizi and possesses remarkable antioxidant and anti-inflammatory capacities. The pharmacological efficacy of Zhizi is highly related to the antioxidant and anti-inflammatory effects of geniposide. Aim of the studyUlcerative colitis (UC) is a common chronic gastrointestinal disease as a global public health threat. Redox imbalance is an essential factor in the progression and recurrence of UC. This study aimed to explore the therapeutic effect of geniposide on colitis and uncover the underlying mechanisms of geniposide-mediated antioxidant and anti-inflammatory activities. Experimental designThe study design involved investigating the novel mechanism by which geniposide ameliorates dextran sulfate sodium (DSS)-induced colitis in vivo and lipopolysaccharide (LPS)-challenged colonic epithelial cells in vitro. Materials and methodsThe protective effect of geniposide against colitis was evaluated by histopathologic observation and biochemical analysis of colonic tissues in DSS-induced colitis mice. The antioxidant and anti-inflammatory effects of geniposide were evaluated in both DSS-induced colitis mice and LPS-challenged colonic epithelial cells. Immunoprecipitation, drug affinity responsive target stability (DARTS), and molecular docking were performed to identify the potential therapeutic target of geniposide and the potential binding sites and patterns. ResultsGeniposide ameliorated the symptoms of DSS-induced colitis and colonic barrier injury, inhibited pro-inflammatory cytokine expression, and suppressed activation of the NF-κB signaling in colonic tissues of DSS-challenged mice. Geniposide also ameliorated lipid peroxidation and restored redox homeostasis in DSS-treated colonic tissues. In addition, in vitro experiments also showed that geniposide exhibited significant anti-inflammatory and antioxidant activity, as evidenced by suppressed IκB-α and p65 phosphorylation and IκB-α degradation, and enhanced the phosphorylation and transcriptional activity of Nrf2 in LPS-treated Caco2 cells. ML385, a specific Nrf2 inhibitor, abolished the protective effect of geniposide against LPS-induced inflammation. Mechanistically, geniposide could bind to KEAP1, thereby disrupting the interaction between KEAP1 and Nrf2, preventing Nrf2 from degradation and activating the Nrf2/ARE signaling pathway, ultimately suppressing the onset of inflammation caused by redox imbalance. ConclusionsGeniposide ameliorates colitis by activation of Nrf2/ARE signaling, while preventing colonic redox imbalance and inflammatory damage, indicating that geniposide can be considered as a promising lead compound for the treatment of colitis.

Administration of necrostatin-1 ameliorates glucocorticoid-induced osteonecrosis of the femoral head in rats.

Glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH) is a serious complication of glucocorticoid treatment and is characterized by dysfunctional bone reconstruction at necrotic sites. Our previous study confirmed the protective potential of necrostatin-1, a selective blocker of necroptosis, in glucocorticoid-induced osteoporosis. In this study, rat models of GC-induced ONFH were established to evaluate the effects of necrostatin-1 on osteonecrotic changes and repair processes. Osteonecrosis was verified by histopathological staining. An analysis of trabecular bone architecture was performed to evaluate osteogenesis in the osteonecrotic zone. Then, necroptotic signaling molecules such as RIP1 and RIP3 were examined by immunohistochemistry. Histopathological observations indicated that necrostatin-1 administration reduced the incidence of osteonecrosis and the osteogenic response in subchondral areas. Additionally, bone histomorphometry demonstrated that necrostatin-1 intervention could restore bone reconstruction in the necrotic zone. The protective mechanism of necrostatin-1 was related to the inhibition of RIP1 and RIP3. Necrostatin-1 administration alleviated GC-induced ONFH in rats by attenuating the formation of necrotic lesions, recovering the function of osteogenesis, and suppressing glucocorticoid-induced osteocytic necroptosis by inhibiting the expression of RIP1 and RIP3.

Ischemic postconditioning protects against acute kidney injury after limb ischemia reperfusion by regulating HMGB1 release and autophagy

Ischemic postconditioning (I-PostC) has a protective effect against acute kidney injury (AKI) induced by limb ischemia–reperfusion (LIR); however, the exact mechanism remains to be elucidated. Our study aims to investigate the potential involvement of high-mobility group box 1 protein (HMGB1) and autophagy in renoprotection generated by I-PostC. A rat model of LIR-induced AKI was established and rats were randomly assigned to five groups: (i) sham-operated control, (ii) I/R, (iii) I/R + I-PostC, (iv) I/R + I-PostC + rapamycin (autophagy activator), and (v) I/R + I-PostC + 3-methyladenine (autophagy inhibitor). Morphological changes in the kidneys were assessed by histology, and ultrastructural changes in renal tubular epithelial cells and glomerular podocytes were observed by transmission electron microscopy. The levels of kidney function parameters, serum inflammatory factors, and autophagy markers were detected. The results showed that the levels of HMGB1, Beclin1, LC3-II/LC3-I, and inflammatory cytokines (TNF-α and IL-6) were significantly higher in the I/R group compared to the sham control in serum and in renal tissues. I-PostC significantly reduced the levels of HMGB1, Beclin1, LC3-II/LC3-I, and inflammatory cytokines in renal tissues and improved renal function. Renal histopathology and ultrastructural observations indicated that I-PostC alleviated renal tissue injury. In addition, rapamycin (autophagy activator) treatment increased the levels of inflammatory cytokine expression levels and decreased renal function, reversed the protective effect of I-PostC against LIR-induced AKI. In conclusion, I-PostC could play a protective role against AKI by regulating the release of HMGB1 and inhibiting autophagy activation.

DMBT1 Alleviates Nasal Airway Inflammatory Response in the LPS-Induced Nasal Polyp Model

Introduction: The aim of this study was to investigate the effects and mechanism of deleted in malignant brain tumors 1 (DMBT1) protein on the mouse model of nasal polyps. Methods: The mouse model of nasal polyps was induced by intranasal drip intervention of lipopolysaccharide (LPS) 3 times a week for 12 weeks. A total of 42 mice were randomly divided into blank group, LPS group, and LPS+DMBT1 group. DMBT1 protein was applied by intranasal drip intervention in each nostril after LPS. After 12 weeks, 5 mice in each group were randomly picked for the mouse olfactory disorder experiment, 3 mice were randomly picked for histopathological observation of nasal mucosa, 3 mice for olfactory marker protein (OMP) immunofluorescence analysis and the last 3 mice were grabbed for nasal lavage, and the levels of cytokines interleukin (IL)-4, IL-5, IL-13, and phosphatidylinositide 3-kinases (PI3K) in the nasal lavage fluid were detected by enzyme-linked immunosorbent assay (ELISA). Results: Compared with the blank group, mice in LPS group had olfactory dysfunction, the level of OMP was significantly reduced, the nasal mucosa was swollen, discontinuous, and contained a large number of inflammatory cells. The levels of IL-4, IL-5, IL-13, and PI3K in the nasal lavage fluid were significantly increased in LPS group (p < 0.01). Compared with the LPS group, the number of mice with olfactory dysfunction in the LPS+DMBT1 group was less, the infiltration of inflammatory cells was reduced, the OMP-positive cells were significantly increased, and the IL-4, IL-5, IL-13, and PI3K in the nasal lavage fluid were significantly increased, p < 0.01. Conclusions: DMBT1 protein alleviates the nasal airway inflammatory response in the mouse nasal polyp model, and the mechanism may be through the PI3K-AKT signaling pathway.

Dynamic Distribution of Mesanophrys sp. and Tissue Enzyme Activities in Experimentally Infected Mud Crab Scylla paramamosain

Mesanophrys sp. is reported to be highly pathogenic to marine crustaceans. This study presents the first report of Mesanophrys sp. infection in the mud crab (Scylla paramamosain). In this study, we first recorded the survival rates of an experimentally infected group and a control group; the cumulative survival rate in the infected group was significantly lower compared to the control group after 72 h (73.20% vs. 94.19%), while the highest mortality of S. paramamosain occurred within the first 24 h post-infection. Then, we investigated the dynamic distribution and tissue tropism of the Mesanophrys sp. in the infected S. paramamosain by a quantitative real-time polymerase chain reaction (qPCR). The result showed that a significant increase in the number of Mesanophrys sp. could be detected in all tested tissues (obtained from the eyestalks, gills, heart, nerves, muscles and hepatopancreas) at 3 h post-infection. The numbers of Mesanophrys sp. in the gill, eyestalk and nerve tissues were relatively higher than in the other tissues. The gill tissue showed the highest numbers from 6 to 48 h. Histopathological observation found a severe collapse in the filament structure, which indicated tissue-specific pathogen infection. Furthermore, the antioxidant enzyme activity of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in three representative tissues (gill, muscle and hepatopancreas) were compared between the infected and control groups, and a significant increase in enzyme activity was observed in all three tested tissues in the infected group, indicating a relatively strong innate immune defense reaction that could have been induced by Mesanophrys sp. infection. These results will be helpful to Mesanophrys sp. pathogenicity-related research and the control of this pathogen in S. Paramamosain in the future.

Evaluation of the biocontrol potential of the fungus Botrytis euroamericana strain HZ-011 for herbicidal activity

BackgroundWeeds in farmland seriously threaten crop yield and cause huge economic losses. Due to the extensive use of chemical herbicides, a series of problems have been arisen, such as environmental pollution, soil degradation and pesticide residues. To assess the herbicidal activity, crop safety, taxonomic identity and infection process of strain HZ-011, the methods of inoculation on detached leaves in vitro and pot plants in vivo, as well as scanning electron microscopy, were used in this study.ResultsThe results indicated that strain HZ-011 had pathogenic effects on detached leaves of four weeds, including Amaranthus retroflexus Linn., Elsholtzia densa Benth., Malva crispa Linn. and Chenopodium album Linn. in vitro. Strain HZ-011 also showed high pathogenicity to C. album and A. retroflexus in vivo, in which the pathogenicity rates were 100%, meaning all plants died after 7 days in the pot test, while the pathogenicity rates for E. splendens and M. crispa were 60.00 and 29.60%, respectively. This strain was safe for local crops, including Vicia faba Linn., Pisum sativum Linn., Brassica napus Linn., Hordeum vulgare Linn. and Triticum aestivum Linn. Strain HZ-011 was identified as the fungus Botrytis euroamericana based on its morphology, molecular biology and a constructed phylogenetic tree. The infection process of B. euroamericana HZ-011 in C. album was studied by plant histopathological observations after pathogen infection. This procedure showed that the mycelium of strain HZ-011 invaded C. album tissues from the stomata, infected and propagated within the tissues, and the spores produced further damage in the C. album tissues and lesions occurred on the surfaces of C. album leaves.ConclusionThese tests provide a basis for fungus B. euroamericana HZ-011 as a potential microbial herbicide.

Efficacy of short-chain polypeptide-based EEN formulas in alleviating intestinal injury in children with Crohn's disease: a single-center study in China.

Short-chain polypeptides are composed of three to nine amino acids, which can be absorbed by the intestinal tract without digestive enzymes and ATP energy. Crohn's disease (CD) is a chronic non-specific disease derived from inflammation and damage of the gastrointestinal tract. In this study, we aim to investigate the effect of short-chain polypeptide-based exclusive enteral nutrition (EEN) formulas on intestinal injury in Chinese children with active CD. From January 2013 to January 2019, a total of 84 consecutive children with a diagnosis of Crohn's disease (CD) in the Department of Pediatric Gastroenterology, Children's Hospital of Nanjing Medical University, were divided into mild and moderate-to-severe active CD groups. Each group was further divided into two subgroups: drug group and short-chain polypeptide plus drug group. Tests were carried out on the levels of intestinal fatty acid binding protein (I-FABP) in the blood, fecal calprotectin (FC), and occludin protein in the intestinal mucosa 1 day before treatment and 8 weeks after treatment. Endoscopic and histopathological observations were detected to compare the changes in intestinal injury in children with active CD. After 8 weeks of treatment, the SES-CD scores and Chiu scores of the ileocecal area and terminal ileum of children with mild active CD and the ileocecal area of children with moderate-to-severe active CD in short-chain polypeptide plus drug group were significantly lower than those in the drug group. The OD value of occludin in the terminal ileum and ileocecal area of children with mild active CD and the ileocecal area of children with moderate-to-severe active CD after short-chain polypeptide-based EEN formulas and drug treatment was significantly higher than those in the drug group (p < 0.05). Meanwhile, the levels of FC and I-FABP were significantly decreased (p < 0.05). The results showed that short-chain polypeptide-based EEN formulas effectively alleviate intestinal injury in children with active CD.

New insight into the toxic effects of lithium in the ragworm Perinereis cultrifera as revealed by lipidomic biomarkers, redox status, and histopathological features.

Lithium (Li) is a toxic monovalent alkaline metal used in household items common to industrial applications. The present work was aimed at investigating the potential toxic effects of LiCl on the redox status, fatty acid composition, and histological aspects of the marine ragworm Perinereis cultrifera. Sea worms were exposed to LiCl graded doses (20, 40, and 80mg/L) for 48h. Compared with the control group, the saturated fatty acids (SFA) decreased while monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) increased upon exposure to LiCl. The increase in PUFA n-3 and PUFA n-6 was concomitant to an increase in docosahexaenoic (DHA: C22:6n-3), eicosapentaenoic (EPA: C20:5n-3), and docosapentaenoic acid (C22:5n-6) fatty acids. Results showed that LiCl-treated specimens accumulate lithium with increasing exposure gradient. Indeed, the exposure to LiCl doses promoted oxidative stress with an increase of the ferric reducing antioxidant power (FRAP), malondialdehyde (MDA), hydrogen peroxide (H2O2), advanced oxidation protein product (AOPP), and protein carbonyl (PCO) as well as the enzymatic and non-enzymatic antioxidants (non-protein thiols (NPSH), catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST), and metallothionein (MT)) levels in all treated groups. Our biochemical findings have been affirmed by the histopathological observations showing hyperplasia and loss of the intestine structure in treated specimens. Overall, our findings give new insights on the toxic effect of LiCl on the redox status of P. cultrifera body tissue and highlighted the usefulness of the FA composition as an early sensitive bioindicators to better understand LiCl mechanism of toxicity in marine polychaetes.

Intestinal inflammation caused by anti-cancer drugs is ameliorated by ginsenoside

Background &amp; Aim: It has been reported that 5-fluorouracil (5-FU), a widely used anticancer drug, causes intestinal inflammation due to shortening of small intestinal villi and destruction of the glandular fossa, resulting in diarrhea and weight loss. Although Ginsenoside Rd (GRd) and Ginsenoside Re (GRe) are saponins found in carrots and have been reported to have anti-inflammatory effects on the gastrointestinal tract, these compounds have not been reported to have anti-inflammatory effects on the small intestinal mucositis induced by 5-FU. In the present study, we investigated the effects of GRd and GRe on the 5-FU-induced mucoenteritis. Materials &amp; Methods: Male C57BL/6N mice (20-25 g) were treated with 5-FU (50 mg/kg i.p.) and GRd (60 or 100 mg/kg p.o.) or GRe (100 mg/kg p.o.) once daily for 7~8 days. 5-FU was dissolved in saline and GRd and GRe were dissolved in 0.5 % CMC. The group receiving saline and 0.5% CMC was designated as the Vehicle group. Body weight and degree of diarrhea (score 0: normal stools, 1: soft stools, 2: severe soft stools, 3: diarrhea, 4: severe diarrhea) were measured daily during the treatment period. The degree of intestinal inflammation was evaluated by histopathological examination using hematoxylin/eosin (H&amp;E) staining, measurement of myeloperoxidase (MPO) activity, and mRNA expression of inflammation-related molecules after the end of drug administration. The expression of NF-κB in the small intestine was examined by immunohistochemical staining. Results: Decreased body weight and exacerbation of diarrhea were observed from the third day of 5-FU administration, and combined administration of GRd or GRe had no effect on these symptoms. Histopathological observations showed that 5-FU administration shortened small intestinal villi and reduced the number of small intestinal crypt, compared to the vehicle group. However, co-administration of GRd and GRe, significantly suppressed the decrease in the number of small intestinal crypts and cells in the crypt by 5-FU treatment and tended to suppress the increase in expression of TNF-α mRNA. MPO activity in the small intestinal mucosa were increased by 5-FU treatment, and the increase was clearly suppressed in the GRd or GRe combination treatment groups. In addition, the expression of NF-κB was clearly increased at 24 hours after the last dose of 5-FU compared with the Vehicle group, and the increase of NF-κB expression by 5-FU administration was clearly suppressed by the concomitant use of GRd. Conclusion: The results obtained in this study suggest that GRd and GRe show anti-inflammatory effect against 5-FU-induced enteritis and that NF-κB is involved in their action. The authors declare no conflict of interest. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

0.5-5% Supramolecular Salicylic Acid Hydrogel is Safe for Long-Term Topical Application and Improves the Expression of Genes Related to Skin Barrier Homeostasis in Mice Models.

As a keratolytic, salicylic acid (SA) can be topically applied in various formulations and doses in dermatology. Supramolecular SA hydrogel, a new SA formulation with higher bioavailability, is developed and commercially available nowadays. However, there still remain concerns that the long-term and continual application of SA at low concentrations may jeopardize the cutaneous barrier properties. To reveal the long-term effects of 0.5-5% supramolecular SA hydrogel on the skin barrier in normal mice models. The 0.5%, 1%, 2%, and 5% supramolecular SA hydrogel or hydrogel vehicle without SA was applied to mice's shaved dorsal skin once per day respectively. Tissue samples of the dorsal skin were harvested on day 14 and 28 of the serial application of SA for histopathological observation and transcriptomic analysis. Following topical supramolecular SA hydrogel therapy with various concentrations of SA (0.5%, 1%, 2%, and 5%) for 14 days and 28 days, there were no obvious macroscopic signs of impaired cutaneous health and no inflammatory or degenerative abnormalities were observed in histological results. Additionally, the transcriptomic analysis revealed that on day 14, SA dramatically altered the expression of genes related to the extracellular matrix structural constituent. And on day 28, SA regulated gene expression profiles of keratinization, cornified envelope, and lipid metabolism remarkably. Furthermore, the expression of skin barrier related genes was significantly elevated after the application of SA based on RNA-seq results, and this is likely to be associated with the PPAR signaling pathway according to the enrichment analysis. Our findings demonstrated that the sustained topical administration of the 0.5-5% supramolecular SA hydrogel for up to 28 days did no harm to normal murine skin and upregulated the expression of genes related to the epidermal barrier.