IL-1β Gene Research Articles

An Aligned-to-Random PLGA/Col1-PLGA/nHA Bilayer Electrospun Nanofiber Membrane Enhances Tendon-to-Bone Healing in a Murine Model.

The challenge of achieving effective tendon-to-bone healing remains a significant concern in sports medicine, necessitating further exploration. Biomimetic electrospun nanomaterials present promising avenues for improving this critical healing process. To investigate the biological efficacy of a novel aligned-to-random PLGA/Col1-PLGA/nHA bilayer electrospun nanofiber membrane in facilitating tendon-to-bone healing. Controlled laboratory study. The bilayer membrane's composition, combining PLGA/Col1 for tendon attachment and PLGA/nHA for bone integration, was examined using scanning electron microscopy, Fourier transform infrared spectroscopy, and mechanical testing. Positioned between the Achilles tendon and bone, its design aimed for harmonious integration with both types of tissue. In vitro, biocompatibility, cell adhesion, and proliferation of the biomaterial were evaluated using live/dead staining and the CCK-8 assay. Collagen secretion and mineralization were measured for 2 cell types. In vivo, tendon-to-bone insertion samples harvested from mice were analyzed: micro-computed tomography assessed bone formation; histological staining evaluated chondrogenesis, tendinogenesis, and the 4-layer structure of the insertion; and biomechanical testing measured insertion strength. Real-time polymerase chain reaction identified genes involved in tendon-to-bone healing, and transcriptome analysis elucidated the underlying cellular and molecular mechanisms. The optimal composition was determined as 10% 3:1 for aligned PLGA/Col1 and 9% 5:1 for PLGA/nHA. Coculture showed minimal cell death, firm cell adherence, and steady proliferation, with PLGA/Col1 enhancing collagen secretion. In vivo, the material promoted bone and cartilage formation and improved tendon-to-bone interface strength. Transcriptome analysis indicated links to TNF and NF-κB pathways and to genes IL-1β, ADAM8, and EGR2. The novel aligned-to-random PLGA/Col1-PLGA/nHA bilayer nanofiber membrane outperformed other materials in both in vitro and in vivo evaluations, significantly enhancing tendon-to-bone healing. It notably improved cartilage and bone formation, tendon maturation, and biomechanical strength at the surgical interface. These effects may be associated with the TNF and NF-κB pathways and with the genes IL-1β, ADAM8, and EGR2. This study introduces a biomimetic nanofiber membrane enhancing tendon-to-bone healing, which is crucial for sports medicine. Its efficacy in improving healing outcomes, including bone and cartilage formation and biomechanical strength, could significantly lower failure rates in surgical procedures such as rotator cuff repair and anterior cruciate ligament reconstruction. This advancement offers promising implications for patient recovery and the effectiveness of surgical interventions in tendon-to-bone injuries.

ASIC1a mediated nucleus pulposus cells pyroptosis and glycolytic crosstalk as a molecular basis for intervertebral disc degeneration.

One of the etiologic components of degenerative spinal illnesses is intervertebral disc degeneration (IVDD), and the accompanying lower back pain is progressively turning into a significant public health problem. Important pathologic characteristics of IVDD include inflammation and acidic microenvironment, albeit it is unclear how these factors contribute to the disease. To clarify the functions of inflammation and the acidic environment in IVDD, identify the critical connections facilitating glycolytic crosstalk and nucleus pulposus cells (NPCs) pyroptosis, and offer novel approaches to IVDD prevention and therapy. By developing keywords search strategy, literature was found and screened using databases such as PUBMED, Google Scholar, Web of Science, China National Knowledge Infrastructure, and others. Hub genes, protein interaction networks, clinical transcriptome data validation, and enrichment analysis were used to further validate relevant biological pathways. It is clear that disc degeneration is associated with apoptosis or pyroptosis, inflammation, and an acidic environment based on literature review. The process of IVDD is intimately associated with pyroptosis, inflammation, and an acidic environment. The precise mechanism may entail the regulation of key genes such NLRP3, ASIC1a, IL1β, TNF-a, and GSDMD. While the acidic environment exacerbated extracellular matrix degradation and promoted cellular senescence and inflammatory factor expression, it was found to be unfavorable for NPCs survival and proliferation. Moreover, NPCs pyroptosis in an acidic environment, the molecular mechanism behind this phenomenon may be connected to ASIC1a mediated Ca + influx. On the other hand, IVDD can be constantly promoted by the interaction between the degenerating disc's acidic and inflammatory environments through "crosstalk" between anaerobic glycolysis and positive feedback. In summary, the inflammatory process in NPCs is made worse by the buildup of glucose brought on by metabolic problems, such as anaerobic glycolytic processes, and pyroptosis caused by excessive glucose may be mitigated by inhibiting endoplasmic reticulum stress. A new therapeutic approach for IVDD will involve using ASIC1a as a regulatory target to enhance the inflammatory environment and decrease the incidence of NPCs pyroptosis. Following this, anaerobic glycolysis will be regulated, lactic acid generation will be reduced, and the degenerative vicious loop will be blocked.

Rhanterium Epapposum Essential Oil and Its Primary Compounds Control Infection, Inflammation, and Serum Electrolyte Imbalance in Mice with Giardiasis.

The present experimental study seeks to evaluate the in vitro and in vivo effects, as well as the potential mechanisms of action, of Rhanterium epapposum essential oil (REE) and its main constituents against Giardia lamblia infection. The analysis of REE was performed using the Gas Chromatography-Mass Spectrometry (GC-MS) detector. The in vitro effects of REE and its main constituents on viability of G. lamblia cysts and the human intestinal epithelial cell line, as well as their effects on plasma membrane permeability, the reactive oxygen species (ROS) generation, and trophozoite adherence were assessed by MTT assay. Effects of oral administration of REE and its main constituents at doses 10 and 20mg/kg on the number of Giardia cysts, the serum level of electrolytes of sodium (Na+) and potassium (K+), as well as the gene expression of inflammatory genes of TNF-α, IL-1β, IL-10, NF-κB p65, and Toll-like receptor 4 (TLR4)) were assessed. The 50% inhibitory concentrations (IC50) value of REE, MC, CP, LN and MTZ against G. lamblia cysts was 26.7, 32.1, 35.3, 45.1, and 42.1µg/mL, respectively; indicated that REE, MC, and CP displayed high antigiardial effects in comparison with MTZ. The obtained 50% cytotoxic concentration (CC50) value of REE, MC, CP, LN, and MTZ on HIEC-6 cells was 279.1, 363.4, 394.2, 478.3, and 422.1µg/mL, respectively. REE and its main compounds significantly (p < 0.001) increased the plasma membrane permeability and ROS generation in Giardia trophozoites, while, reduced the Giardia trophozoites adherence. In vivo, REE, MC, CP, and LN mainly at the dose of 20mg/kg considerably reduced the number and viability of Giardia cysts, modulated the serum level of electrolytes Na and K in the infected mice compared with the mice treated with MTZ (P < 0.01); whereas inhibited the inflammation cytokines in the infected mice compared with the mice treated with MTZ (p < 0.001). The current investigation showed that R. epapposum essential oil and its main compounds controlled the Giardia infection in mice by inhibiting inflammation, and serum electrolyte imbalance. After confirmation in further studies, these compounds may be considered for development as a novel therapeutic option for the treatment of giardiasis.

P0697 Ozanimod Inhibits Pro-Inflammatory Mediators of the IL-17 Pathway in the Tissues of Participants in the Phase 2 TOUCHSTONE Study

Abstract Background OZA was approved for the treatment of moderately to severely active ulcerative colitis (UC). IL-17A, a proinflammatory cytokine, sustains the release of other inflammatory mediators and is associated with inflammatory bowel disease (IBD). Methods This analysis assessed gene expression from FFPE colon tissue in baseline and post treatment samples during TOUCHSTONE study. 197 Pts were randomized 1:1:1 to double-blind treatment with once-daily oral OZA 0.5 mg and 1 mg or matching placebo (PBO) for a 1-week (W) dose escalation period followed by 8W of induction treatment. Pts with clinical response at W8 (after dose escalation) entered a maintenance phase through W32. A NanoString whole transcriptome atlas (WTA) assay was run on these FFPE colon samples stained with antibodies to S1P1, panCK, and CD45. Six regions of interests (ROIs) were selected in each sample based on the staining and each ROI were split into two areas (CD45+ or CD45-). Differential enriched pathways analysis including IL-17 signaling pathway gene expression was analyzed in colon tissue samples collected before treatment, W8 and W32. Results OZA, not Placebo (PBO), induced significant PD spatial gene expression changes over time in colon tissue. No significant PD effects on gene expression changes were observed in CD45+ or Rest segment in PBO. OZA 0.5 mg induced significant spatial whole transcriptome gene expression changes at week 32 in the CD45+ segment (n=336 genes) or Rest (n=267 genes), and no differential expression genes were observed at week 8. OZA 1mg induced the most significant spatial whole transcriptome gene expression changes as early as W8 in segment CD45+ (n=121 genes) or Rest (n=171 genes). Significant overlap in OZA-included PD gene expression changes across doses, visits or cell types. OZA significantly downregulated 24 pathways in colon tissue from the CD45+ segment at W8 (p&amp;lt;0.05), with IL-17 signaling as the most significantly downregulated pathway. Significant IL-17 related gene differences were identified with OZA treatment, including CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, MMP1, MMP3, MMP9, S100A8, S100A9, LCN2, CEBPB, HSP90B1 and IL1B genes, which involved in inflammation and immune response. Conclusion OZA led to circulating IL-17A reductions (previously reported) and IL-17 pathway related gene expression reduction in colon tissue, indicative of decreases in inflammatory responses. These finding may help support the use of supporting IL-17A and IL-17 pathway related genes as a good PD biomarker for OZA in UC.

Anti-inflammatory Effects of the Fucoidan from Sea Cucumber Apostichopus japonicus.

Fucoidan from Apostichopus japonicus (Aj-FUC) has shown anti-inflammatory activity, whereas its mechanism was not explicated. This study investigated the anti-inflammatory potential and mechanism of the fucoidan from green and purple A. japonicus (G-FUC and P-FUC) in lipopolysaccharide (LPS)-treated RAW264.7 cells. Results showed that Aj-FUCs at 25-400µg/mL had no toxicity to cells after 24h stimulation and promoted cell phagocytic activity. ELISA results indicated that Aj-FUC reduced the nitric oxide (NO), tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 levels and increased IL-10 level. The Aj-FUC suppressed transcription of inflammatory-related genes (tnf-α, il-1β, il-6, nlrp3, inos, cox-2, tlr4, trif, and nf-κb) in LPS-treated RAW264.7 cells, among which G-FUC had stronger anti-inflammatory effects. Moreover, Aj-FUC upregulated the mRNA expression of autophagic genes (beclin1, lc3II, and lamp2). The immunoblotting and immunofluorescence analyses of Beclin-1 and LC3II supported that Aj-FUC enhanced autophagy activity. After autophagy inhibited by 3-methyladenine, the mRNA expressions of tnf-α, il-6, il-1β, and nlrp3 were significantly upregulated in LPS-induced cells treated with Aj-FUC, suggesting the suppressed inflammation by Aj-FUC mediated via autophagy. Summarily, the present study demonstrated that Aj-FUC showed anti-inflammatory effects by elevating autophagy activity in LPS-induced macrophages.

The Role of TSLP and IL-1 β and Their Genetic Variants in the Pathogenesis of Single and Multiple Atopic Diseases in Children.

Allergic diseases commonly coexist, manifesting in a sequence described as the "allergic march". Background/Objectives: This study aimed to evaluate TSLP's and IL-1β's potential as biomarkers in both single and multi-pediatric atopic diseases like atopic eczema, food allergy, and anaphylaxis and analyze specific SNPs in the TSLP and IL-1β genes to determine their associations with their occurrence and severity. Methods: This analysis included 109 atopic children diagnosed with atopic dermatitis, food allergy, or anaphylaxis alongside a control group of 57 non-atopic children. Recruitment was facilitated through the use of a comprehensive questionnaire. For the study population, the allergen profile was characterized at the molecular level by measuring specific IgE to purified natural or recombinant allergens, assessing serum levels of circulating TSLP and IL-1β, and identifying single-nucleotide polymorphisms in TSLP (rs2289277) and IL-1β (rs16944 C-511T). Results: The serum levels of TSLP and IL-1β were elevated in the study groups compared to the control group, highlighting their significance in the pathogenesis of the studied diseases. Carrying a higher number of the risk allele [C] in the TSLP SNP rs2289277 is associated with the greatest likelihood of having multiple concurrent allergic conditions, with the highest risk observed in individuals with all three conditions-atopic dermatitis, food allergy, and anaphylaxis, simultaneously. Moreover, children carrying the risk allele had a twofold increased risk of polysensitization, which rose to sixfold in those with two copies of the risk allele. Although no significant variations in genotype frequencies were detected for IL-1β rs16944, significant associations were observed for TSLP rs2289277, particularly with conditions such as atopic dermatitis, food allergy, anaphylaxis, and combinations of these diseases. Conclusions: Further research is required to elucidate these pathways and their role in the development of allergic diseases.

Integrative RNA, miRNA, and 16S rRNA sequencing reveals immune-related regulation network for glycinin-induced enteritis in hybrid yellow catfish, Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂

Glycinin-induced foodborne enteritis is a significant obstacle that hinders the healthy development of the aquatic industry. Glycinin causes growth retardation and intestinal damage in hybrid yellow catfish (Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂), but its immune mechanisms are largely unknown. In the current study, five experimental diets containing 0% (CK), 1.74% (G2), 3.57% (G4), 5.45% (G6), and 7.27% (G8) immunological activity of glycinin were fed to juvenile hybrid yellow catfish to reveal the mechanism of the intestinal immune response to glycinin through RNA and microRNA (miRNA) sequencing and to explore the interrelation between immune molecules and intestinal microbiota. The results demonstrated that glycinin content in the posterior intestine increased significantly and linearly with the rise of dietary glycinin levels. More than 5.45% of dietary glycinin significantly reduced the nutritional digestion and absorption function of the posterior intestine. Notably, an obvious alteration in the expression levels of inflammatory genes (tnf-α, il-1β, il-15, and tgf-β1) of the posterior intestine was observed when dietary glycinin exceeded 3.57%. Sequencing results of RNA and miRNA deciphered 4,246 differentially expressed genes (DEGs) and 28 differentially expressed miRNAs (DEmiRNAs) between the CK and G6 groups. Furthermore, enrichment analysis of DEGs and DEmiRNA target genes exhibited significant responses of the MAPK, NF-κB, and WNT pathways following experimental fish exposure to 5.45% dietary glycinin. Additionally, at the level of 3.57% in the diet, glycinin obviously inhibited the increase of microbiota, especially potential probiotics such as Ruminococcus bromii, Bacteroides plebeius, Faecalibacterium prausnitzii, and Clostridium clostridioforme. In sum, 5.45% dietary glycinin through the MAPK/NF-κB/WNT pathway induces enteritis, and inflammatory conditions could disrupt micro-ecological equilibrium through miRNA secreted by the host in hybrid yellow catfish. This study constitutes a comprehensive transcriptional perspective of how intestinal immunity responds to excessive glycinin in fish intestines.

Evaluating the effects of c-phycocyanin from cyanobacterium Spirulina platensis as an anti-inflammatory agent on stimulated human chondrocyte cells

ABSTRACT Inflammation is a critical defence mechanism in higher animals but uncontrolled inflammation is implicated in many diseases and can lead to chronic conditions, posing significant challenges to healthcare and the global economy. Conventional therapies often come with limitations and adverse effects, underscoring the need for safer and more effective alternatives. This study investigates the efficacy of C-Phycocyanin (C-PC), a pigment derived from Spirulina platensis cultivated under optimal light conditions, as an anti-inflammatory agent. This research focused on its effects on nitric oxide secretion, which supports the body’s immune system by killing pathogens, and interleukin-1 beta (IL-1β) gene expression as an inflammatory indicator, in phorbol 12-myristate 13-acetate (PMA)-stimulated human chondrocyte cells (C28/I2). The Methyl Thiazol Tetrazolium (MTT) assay was employed to evaluate the metabolic activity of cells and confirm the non-toxicity of C-PC. The results demonstrated that C-PC maintained 99.32% cell viability in the C28/I2 in vitro model after seven days. Notably, C-PC treatment prevented cell death and promoted cell proliferation, increasing the number of viable cells by 12.34% during the same period. Additionally, it significantly reduced IL-1β gene expression and nitric oxide secretion by 70.24% and 91.25%, respectively, effectively reducing levels to those observed in unstimulated conditions. Scanning electron microscopy (SEM) and crystal violet staining showed that C-PC treatment restored normal morphology and protected the cells from inflammation-induced changes.

Study on the Role and Pathological and Immune Responses of Silver Nanoparticles Against Two Aeromonas salmonicida subsp. salmonicida Strains at Different Virulence Levels in Rainbow Trout (Oncorhynchus mykiss)

Aeromonas species are among the main pathogens causing rainbow trout infections. Silver nanoparticles (AgNPs) have a broad spectrum of antimicrobial properties and are usually produced by various green-synthesis methods. However, the application of commercialized AgNPs has not fully been clarified. Thus, the objective of this study was to evaluate the antibacterial activities of commercialized AgNPs (range of sizes 10–12 nm) on two contrasting A. salmonicida strains (I-1 and I-4), isolated from rainbow trout; the antibacterial mechanism, histopathological alterations and the expression of immune-related genes were investigated. In vitro, the minimal inhibitory concentration (MIC) was 10 µg/mL for I-1, and lowered to 9.5 µg/mL for I-4, respectively. AgNPs were shown to disrupt both the cell wall and membrane of I-1 and I-4, resulting in cell lysis and degradation. In vivo, rainbow trout challenged by immersed or intraperitoneally injected infection, the 10 µg/mL AgNP-treated groups, both showed delayed deaths and lower mortalities compared to the control groups, without any clinical signs and pathological changes. Especially for the virulent I-4, the enhanced expressions of immune-related genes TNF-α, IL-1β, IL-10 and IL-11 were significantly reduced in the AgNP-treated group, indicating a lesser inflammation due to the application of AgNPs. This study would lay theoretical foundation for the wide application of silver nanoparticles in fish diseases.

Effects of Dietary Nano-Composite of Copper and Carbon on Antioxidant Capacity, Immunity, and Cecal Microbiota of Weaned Ira White Rabbits.

This experiment investigated the effects of dietary supplementation with nano-composites of copper and carbon (NCCC) on antioxidants, immune functions, and the cecum microbiota of weaned Ira white rabbits. A total of 240 weaned 35-day-old Ira white rabbits were randomly allocated to five dietary treatments (n = 6 per treatment, each replicate consisted of eight rabbits) that included the control group (CON) with a basal diet, the SAL group with 60 mg/kg salinomycin (SAL) in addition to the basal diet, and the NCCC I, II, III groups, which were supplemented with 50, 100, and 200 mg/kg NCCC, respectively, in addition to the basal diet. The test lasted for 28 d. The results showed that dietary NCCC supplementation increased the liver Cu/Zn-SOD content and up-regulated the gene expression of Cu/Zn-SOD (p < 0.05), while also reducing the content of MDA in the liver and enhancing the antioxidant capacity of Ira white rabbits. Moreover, the NCCC diet supplementation reduced the content of IL-6 and down-regulated the relative expression of IL-6 and IL-1β genes in the jejunum of Ira white rabbits (p < 0.05). In addition, the metagenomic analysis of 16 S rRNA showed significant differences in the cecal microbial structure of weaned Ira white rabbits in the NCCC III group compared with the CON, NCCC I, and NCCC II groups (p < 0.05). Firmicutes and Bacteroidetes were the dominant phyla of cecal microorganisms in weaned Ira rabbits in the NCCC diet groups. The dominant genera included unidentified Eubacteriaceae, unclassified Lachnospiraceae, Christensenellaceae, and Ruminococcus. Furthermore, the relative abundance of Ruminococcus in the NCCC I and II groups was lower than that in the CON group in the cecum of Ira white rabbits (p < 0.05). In summary, our results showed that diet supplementation with NCCC could enhance the antioxidant capacity in the liver, alleviate intestinal inflammation, and regulate the structure of intestinal flora, improving the health of Ira white rabbits.

New insight into the role of the pathway NLRP1 and NLRP3 inflammasomes and IL-33 in ultraviolet-induced cutaneous carcinogenesis.

Inflammasomes NLRP1 (NLR family pyrin domain containing 1) and NLRP3 are pivotal regulators of the innate immune response, activated by a spectrum of endogenous and exogenous stressors, including ultraviolet radiation (UVR). The precise molecular mechanisms underlying the activation of these inflammasomes remain unclear. Furthermore, the involvement of interleukin-33 (IL-33) in UVR-induced skin carcinogenesis is not well defined. The objective of this study is to evaluate the expression of interleukin genes (IL-33, IL-18, IL-1β) following the activation and silencing of NLRP1 and NLRP3 at various wavelengths and doses of UV radiation, and to correlate these expressions with pertinent tumor markers (e.g., Gli1, Gli2, FOXO3A, SerpinA1, SerpinA3, and EphB2). Cultures of keratinocyte cell lines were exposed to varying doses of UV radiation using specific lamps. To inhibit the expression of NLRP1 and NLRP3 genes, cells were transfected with targeted siRNAs. Gene expression of inflammasome components and effector proteins was quantified using Real-time PCR and ELISA. There was a marked upregulation in the expression levels of cytokine genes IL-18, IL-1β, and IL-33 upon exposure to UVB and UVA radiation, compared to non-irradiated keratinocytes. Silencing NLRP1 or NLRP3 via RNA interference in primary human keratinocytes resulted in a significant reduction of cytokine gene expression. Additionally, a notable increase in tumor marker gene expression was observed in cells with functional NLRP1 and NLRP3 following UV radiation, whereas silencing these inflammasome genes altered the expression profiles of these markers. This study provides a pioneering comprehensive assessment of the roles of NLRP1, NLRP3, and IL-33 in the pathogenesis of UV-induced cutaneous carcinogenesis. Our findings substantiate the role of IL-33 as a critical early danger signal elicited in response to inflammatory UV radiation, presumably regulated by inflammasomes.

Sea Anemone Kunitz Peptide HCIQ2c1 Reduces Histamine-, Lipopolysaccharide-, and Carrageenan-Induced Inflammation via the Suppression of Pro-Inflammatory Mediators

Inflammation is a physiological response of the immune system to infectious agents or tissue injury, which involves a cascade of vascular and cellular events and the activation of biochemical pathways depending on the type of harmful agent and the stimulus generated. The Kunitz peptide HCIQ2c1 of sea anemone Heteractis magnifica is a strong protease inhibitor and exhibits neuroprotective and analgesic activities. In this study, we investigated the anti-inflammatory potential of HCIQ2c1 in histamine- and lipopolysaccharide (LPS)-activated RAW 264.7 macrophages as well as in LPS-induced systemic inflammation and carrageenan-induced paw edema models in CD-1 mice. We found that 10 μM HCIQ2c1 dramatically decreases histamine-induced intracellular Ca2+ release and LPS-induced reactive oxygen species (ROS) production in RAW 264.7 macrophages. Moreover, HCIQ2c1 significantly inhibited the production of LPS-induced tumor necrosis factor α (TNF-α), inducible NO-synthase (iNOS), and 5-lipoxygenase (5-LO) but slightly influenced the IL-1β and cyclooxygenase-2 (COX-2) expression level in macrophages. Furthermore, intravenous administration by HCIQ2c1 at 0.1 mg/kg dose reduced LPS-induced TNF-α, IL-1β, COX-2, and iNOS gene expression in CD-1 mice. The subplantar administration of HCIQ2c1 at 0.1 mg/kg dose to mice significantly reduced carrageenan-induced paw edema by a factor of two, which is comparable to the effect of diclofenac at 1 mg/kg dose. Thus, peptide HCIQ2c1 has a strong anti-inflammatory potential by the attenuation of systemic and local inflammatory effects through the inhibition of intracellular Ca2+ release, the production of ROS and pro-inflammatory cytokines, and enzymes involved in arachidonic acid metabolism.

Bacillus coagulans alleviates hepatic injury caused by Klebsiella pneumoniae in rabbits.

As an opportunistic bacterial pathogen, Klebsiella pneumoniae (KP) is prone to causing a spectrum of diseases in rabbits when their immune system is compromised, which poses a threat to rabbit breeding industry. Bacillus coagulans (BC), recognized as an effective probiotic, confers a variety of benefits including anti-inflammatory and antioxidant properties. The purpose of this study was to investigate whether dietary BC can effectively alleviate hepatic injury caused by KP. In this study, the rabbits were initially pretreated with varying doses of BC (1×106, 5×106, and 1×107 CFU/g), followed by a challenge with KP at a concentration of 1011 CFU/mL. Liver tissues were harvested and processed for histological assessment using H&E and VG stains to assess structural alterations. Biochemical assays were employed to quantify the enzymatic activities of T-SOD and GSH-Px, as well as the MDA content. Furthermore, ELISA was utilized to detect the levels of inflammatory cytokine (IL-10, IL-6, IL-1β and TNF-α) and apoptotic-related gene (Bcl-2, Bax). Morphological observation indicated that BC can effectively mitigate KP-induced hepatic sinusoidal dilatation and congestion, as well as ameliorate the degree of hepatic fibrosis. Further analysis showed that BC significantly lowered MDA level in KP-treated rabbits, while enhanced the activities of T-SOD and GSH-Px. Additionally, ELISA result showed that BC pretreatment significantly reduced the levels of pro-inflammatory cytokines TNF-a, IL-6, IL-1β and pro-apoptotic gene Bax, while increasing the levels of anti-inflammatory cytokine IL-10 and anti-apoptotic gene Bcl-2 in KP-treated rabbits. Above data indicate that BC supplementation effectively attenuated oxidative stress and inflammatory response induced by KP through augmenting the activities of antioxidant enzymes and diminishing the levels of pro-inflammatory factors. Furthermore, it reduced the Bax/Bcl-2 ratio in the liver, thereby inhibiting KP-induced apoptosis. The treatment group receiving 5x106 CFU/g BC benefitted most from the protective effect.

Evaluation of bone development and organs in rat fetuses exposed to tartrazine.

Tartrazine finds widespread application in the realms of alimentation, pharmaceuticals, cosmetic formulations, and textile manufacturing. Tartrazine has a negative effect on human health such as hyperactivity, allergies and asthma in children. Substances such as tartrazine might effect the embryo in a kind of aspects, containing physical or mental disorders, and a decrease in the child's intellectual memory. In this study, Sprague dawley female rats, 70-100 days old, weighing 250-300g, with confirmed pregnancy, were divided into two groups of 5: control and tartrazine group. Rats were sacrificed on the 20th day and heart, lung, kidney and liver tissues were removed from the fetuses. The effect of tartrazine on fetal bone development was assessed by double skeletal staining, histological analysis on organs, and IL-6, IL-1β, TNF-α, and TRPM2 gene levels. It has been observed that tartrazine, which is frequently used as a food dye, damages important fetal tissues such as liver, kidney, lung and heart. A statistically meaningful reduce was observed in the total length, total area, bone length and bone area values of the limb bones in the tartrazine group compared to the control group (p<0.05). It was observed that the mRNA levels of IL-6, IL-1β, TNF-α and TRPM2 genes in the livers of the fetuses changed compared to the control group (p<0.05). In this study on the use of tartrazine during pregnancy, it was observed that both organs and bone development were damaged. More studies are needed on the effects of tartrazine.

Temperature-dependent toxicity and mechanisms of florfenicol on the embryonic development of marine medaka (Oryzias melastigma).

The extensive use of antibiotics and their persistence in the environment have seriously threatened marine ecosystems in recent years. The frequent occurrence of extreme weather due to climate change has also increased the uncertainty of effective toxicity identification and risk assessment of the chemicals of concern. This study aimed to investigate the toxic effects and potential mechanisms of florfenicol (0.5, 10, 100, 500 and 1000 μg/L) on the embryonic development of marine medaka (Oryzias melastigma) through 21-d experiment under three temperature exposure scenarios (20, 25, and 30 °C). Considering embryo development, the highest level of florfenicol at 1000 μg/L decreased the hatching success at 25 °C, whereas the total inhibition of hatching was observed at 20 °C regardless of florfenicol concentrations of concern. The ATP content was inhibited by the highest florfenicol dose at 20 °C, while stimulated ATP content at 20 °C and 30 °C, compared to 25 °C. Fluctuation of temperatures, especially at 20 ℃, induced oxidative stresses, including increased MDA contents and decreased CYP450 and HSP90 contents. For inflammatory response-related genes (nf-κb, nlrx1, pycard, caspase 1, and il-1β), an increase of florfenicol dose led to gene upregulation at 25 °C. Conversely, gene upregulation was also observed for all treatments at 30 °C, while predominantly downregulation was observed for all treatments at 20 °C. For genes related to DNA damage and apoptosis (pi3k, akt, foxo1, tp53, bcl-2, bax, apaf-1, caspase 9, and 3), alterations in gene expression were evident for all florfenicol treatments at both 20 °C and 30 °C. Therefore, this study suggests that the combined effects of florfenicol and temperature may disrupt the normal function of mitochondria, impacting ATP production, thereby inducing oxidative stress, inflammatory response, DNA damage, and apoptosis, ultimately resulting in decreased hatching rates and increased mortality. Furthermore, the key findings of this study reveal the complex interactions between florfenicol and temperature and emphasize the need to consider temperature when identifying toxicity and mechanisms, as well as the ecological risk assessment of florfenicol in marine environments.

Molecular genetic characteristic of pulmonary tuberculosis associated with ABCB1 gene expression of multidrugresistance protein P-gp

Background: Tuberculous inflammation is mediated by a complex molecular signaling pathway, the analysis of which makes it possible to identify promising biomarkers and targets for the development of new diagnostic, prognostic and pharmacological approaches in order to improve the effectiveness of anti-tuberculosis chemotherapy. Determining the relationship between key inflammatory cytokines, the multidrug-resistant protein P-gp and the activity of specific inflammation in the surgical material of patients with pulmonary tuberculosis may prove to be a novel tool in the development of pathogenetic therapy and personalized medicine. Aims: to characterize molecular and genetic profiles of tuberculomas and identify genes that correlate with the expression of the ABCB1 gene of the P-gp protein in the surgical material of patients with pulmonary tuberculosis. Research objectives: 1) to obtain molecular and genetic characteristics of tuberculosis by real-time PCR and compare it with the activity of tuberculous inflammation; 2) to carry out a correlation analysis between the expression of the ABCB1 gene and key cytokines of the tuberculosis process: IL-6, IL-10, IFN-γ, TGF-β, TNF-α, IL-1β. Materials and methods: A prospective cohort study was conducted on the basis of the FSBI CTRI. The object of the study was the surgical material of 35 patients diagnosed with multiple pulmonary tuberculomas. Histological examination methods were used for the morphological assessment of the surgical material. A real-time quantitative PCR method was used to analyze gene expression. Statistical processing was performed using the GraphPad Prism Version 7.04 software package (GraphPad Software, USA). The data is presented as a median with an interquartile range. The nonparametric Mann-Whitney U-test was used to compare the two groups. All p-values were two-sided and p 0.05 was considered statistically significant. The correlation between the variables was estimated using the Spearman correlation coefficient. The correlation analysis was carried out in the Microsoft Office Excel 2010 software. Results: The study revealed that the highest level of expression of ABCB1 gene of the P-gp protein is observed in tuberculomas with high activity of tuberculous inflammation, and its expression is correlated with the expression of the IL6 gene (p0.001) and the expression of the IL10 gene (p0.01). Tuberculomas of this group are also characterized by higher expression of the TGFB1, TNF and IL1B genes, compared with the group of moderate activity of specific inflammation. Conclusions: The data obtained indicate that in addition to pro-/anti-inflammatory cytokines, the P-gp protein plays an important role in the pathogenesis of tuberculous inflammation, especially with its high activity. Further clarification of the P-gp role in tuberculous inflammation may be an important step for the development of new approaches to treat tuberculosis using methods of HDT and personalized medicine.

The Expressions of the Immunity- and Muscle Development-Related Genes of 40-Day-Old Broilers Are Promoted in Response to the In Ovo and Dietary Supplemental Administration of Calcidiol in Conjunction with the In Ovo Administration of Marek's Disease Vaccine.

Effects of in ovo and dietary sources of calcidiol (25(OH)D3), combined with Marek's disease vaccine (MDV), on the expression of genes involved with the antioxidant activity, muscle deposition, and immunity in the pectoralis major (P. major) muscle and spleen of 40 d of age (doa) broilers were investigated. The in ovo treatments were as follows: (1) non-injected; (2) the injection of 50 μL of commercial MDV, (3) MDV + 1.2, or (4) 2.4 μg of 25(OH)D3. All birds received either a commercial diet containing no supplemental 25(OH)D3 (control) or the same diet supplemented with an additional 69 µg of 25(OH)D3 per kg of feed (Hy-D diet). At 40 doa, the pectoralis major (P. major) muscle and spleen of 48 birds (six replicates per diet x in ovo treatment combination) were collected. When compared to un-supplemented commercial diet-fed birds, in birds that were fed the Hy-D diet, the expression of the TGF-β4 gene in the spleen and P. major muscle, and the GSH-P1, GSH-P7, SOD2, MyoG, MyoD1, and Pax3 genes in the P. major muscle were up-regulated, whereas the expression of the IL-1β, IL-8, and CYP24A1 genes in the spleen and P. major muscle were down-regulated. Nevertheless, birds that received any of the in ovo injection doses of 25(OH)D3 exhibited a higher expression of the IL-10, TGF-β4, and CYP27B1 genes in the spleen and P. major muscle. Furthermore, in comparison to the MDV-injected control group, the CAT, MyoD1, and Pax3 genes in the P. major muscle were up-regulated, and the expression of the INF-γ, IL-1β and CYP24A1 genes in the spleen and the IL-8, and IL-1β genes in the P. major muscle were down-regulated. In conclusion, a significant improvement in the expression of genes responsible for enzymatic antioxidant activity, protein synthesis, and inflammatory reactions in 40-day-old broilers occurred in response to in ovo and dietary supplemental 25(OH)D3, and supplemental 25OHD3 provided via either route was used to enhance the expression of genes linked to vitamin D activity (CYP27B1, CYP24A1).

Mechanism of Liraglutide Regulating the Expression of PI3K/AKT/NF-KB Pathway-related Proteins on Nonalcoholic Fatty Liver Disease

Background and Objective: GLP-1RAs can effectively prevent hepatic steatosis. It was to investigate the mechanism by which the GLP-1RA Liraglutide ameliorates non-alcoholic fatty liver disease (NAFLD) and its impact on PI3K/AKT/NF-κB pathway. Materials and Methods: eight-week-old, clean-grade male C57BL/6J mice were assigned to Normal group (NG, fed standard chow), NAFLD group (high-calorie, high-cholesterol), and Liraglutide group (LG, high-calorie, high-cholesterol for 12 weeks + Liraglutide treatment at 1 mg/kg). Serum samples were collected from the mice to assess changes in glucose and lipid metabolism and liver function (LF) indicators. Histopathological changes in liver tissues (LTs) were visualized. Real-time quantitative PCR measured expression levels (ELs) of inflammatory genes in LTs. Western blotting was conducted to detect EL of proteins related to PI3K/AKT/NF-κB pathway in LTs. Results: NAFLD group exhibited markedly elevated levels of blood glucose (BG), blood lipids, and LF indicators relative to NG. Histopathological examination revealed increased scores for steatosis, lobular inflammation, and hepatocellular ballooning. The ELs of inflammatory genes IL-1β, TNF-α, and NF-κB p65 in LTs, were greatly elevated (P&lt;0.05). In contrast, relative to NAFLD group, LG showed notably reduced levels of BG, blood lipids, and LF indicators, along with decreased histopathological scores. Furthermore, the ELs of inflammatory genes and protein ELs of p-PI3K, p-AKT, and p-NF-κBp65 in LTs were drastically lowered (P&lt;0.05). Conclusion: Liraglutide ameliorates glucose and lipid metabolism disorders and suppresses inflammatory responses in liver by inhibiting PI3K/AKT/NF-κB signaling, thereby improving NAFLD in mice.

Evaluation of the effects of Andiz extract on the experimentally wound in rats by microbiological and gene expression methods.

This study aimed to investigate the effects of andiz extract on wound healing and compare it with saline and chlorhexidine gluconate. Microbial DNA load was used to evaluate its antibacterial effects, and gene expression methods were used to assess its contribution to cytokine release and wound healing. A standardized wound site was created with a 3mm diameter punch on 32 male Wistar albino rats. The rats were divided into four groups: Control (n = 5), Saline (n = 9), Chlorhexidine gluconate (n = 9), and Andiz extract (n = 9). Five rats in the control group were euthanised without any treatment. Irrigations of the Saline, Chlorhexidine, and Extract groups were provided regularly. After the tissue samples were taken in the 1st week, 2nd week, and 3rd week, three rats were euthanized each week for each group. The total bacterial DNA load on the samples taken was determined by a nano spectrophotometer. β-actin was chosen as housekeeping, and target gene primers were created for TGF-β and IL-1β. Expression amounts of target genes were measured by Real-Time PCR with the application of the created primers. There is a significant difference between the Extract group and the other groups regarding total bacterial DNA load. The whole bacterial load was 185% less than the initial values. TGF-β and IL-1β genes evaluated regarding gene expression were measured at the highest value in the Extract group. This study showed the antibacterial effects of the Extract and its positive contributions to wound healing.

Aged Garlic Extract (AGE) and Its Constituent S-Allyl-Cysteine (SAC) Inhibit the Expression of Pro-Inflammatory Genes Induced in Bronchial Epithelial IB3-1 Cells by Exposure to the SARS-CoV-2 Spike Protein and the BNT162b2 Vaccine.

Garlic (Allium sativum L.) is a species of the onion family (Alliaceae) widely used as a food and a folk medicine. The objective of this study was to determine the effects of AGE (aged garlic extract) on pro-inflammatory genes relevant to COVID-19. To this aim, we treated bronchial epithelial IB3-1 cells with SARS-CoV-2 spike protein (S-protein) or with the COVID-19 BNT162b2 vaccine in the absence or in the presence of AGE. The results obtained demonstrated that AGE is a potent inhibitor of the S-protein-induced expression of the IL-1β, IL-6 and IL-8 genes. Bio-Plex analysis demonstrated that AGE reduced release of IL-6 and IL-8, which were highly induced by S-protein. No inhibition of cells' growth, toxicity and pro-apoptotic effects were found in AGE-treated cells. The effects of one of the major AGE constituents (S-allyl cysteine, SAC) were studied on the same experimental model systems. SAC was able to inhibit the S-protein-induced expression of IL-1β, IL-6 and IL-8 genes and extracellular release of IL-6 and IL-8, confirming that S-allyl-cysteine is one of the constituents of AGE that is responsible for inhibiting S-protein-induced pro-inflammatory genes. Docking experiments suggest that a possible mechanism of action of SAC is an interference with the activity of Toll-like receptors (TLRs), particularly TLR4, thereby inhibiting NF-κB- and NF-κB-regulated genes, such as IL-1β, IL-6 and IL-8 genes. These results suggest that both AGE and SAC deserve further experimental efforts to verify their effects on pro-inflammatory genes in SARS-CoV-2-infected cells.