Lung Tissue Research Articles

In-Depth Analysis of Olea europaea L. Leaf Extract: Alleviating Pulmonary Histological Disturbances, Pro-Inflammatory Responses, and Oxidative Stress from Isolated or Combined Exposure to Inhaled Toluene and Noise in Rats

The primary objective of this study was to investigate the pulmonary damage resulting from isolated or combined exposure to inhaled toluene (300 ppm) and noise 85 dB (A), with a focus on evaluating the potential protective effects of Olea europaea L. leaf extract (OLE). Forty-eight male Wistar rats were divided into eight groups: control (C), OLE treatment (O), noise exposure (N), noise exposure with OLE treatment (N+OLE), toluene exposure (T), toluene exposure with OLE treatment (T + OLE), co-exposure to toluene and noise (NT), and co-exposure with OLE treatment (NT + OLE). OLE (40 mg/kg/day) was administered daily for six weeks via oral gavage. Exposure to toluene and noise resulted in significant disruption of the pulmonary tissue structure, accompanied by oxidative stress, as evidenced by increased lipid peroxidation, diminished catalase and superoxide dismutase activities, and elevated pro-inflammatory cytokines IL6, IL-β, and TNF-α. Notably, the administration of OLE effectively mitigated oxidative stress and inflammation and preserved pulmonary histology. In conclusion, exposure to toluene and its combination with noise significantly elevated oxidative stress, inflammatory responses, and histological disruptions in the lung tissue. In contrast, noise exposure alone is characterized by minimal effects, although it is still associated with an inflammatory response. Notably, Olea europaea L. leaf extract (OLE) exhibits a substantial protective role, effectively mitigating the adverse effects of combined exposure and highlighting its potential as a therapeutic agent for lung health.

Analisis Minimalisasi Biaya Terapi Antibiotik Ceftriaxone dan Cefotaxime Pada Pasien Pneumonia di Instalasi Rawat Inap RSUD Kabupaten Kediri

Pneumonia is an infection caused by pathogenic microorganisms such as microbes, bacteria, fungi and viruses that cause inflammation of the lung tissue. The prevalence of pneumonia has increased every year. The first-line therapy used at the Kediri District Hospital is the use of ceftriaxone and cefotaxime antibiotics. Decision making on antibiotic use has an influence on the amount of medical costs incurred by patients. This study aims to determine which antibiotics are more cost-minimal and whether there is a significant difference between the use of ceftriaxone and cefotaxime antibiotic therapy at Kediri District Hospital. The design of this study was cross sectional, data collection was done retrospectively. The analysis method used independent t-test statistical test. The perspective in this study was seen through the provider, namely the hospital, a total of 215 pneumonia patients and the sample for this study was 63 patients. With 32 patients receiving ceftriaxone antibiotic therapy and 31 patients receiving cefotaxime antibiotic therapy. Total direct medical costs were categorized into four types of costs, namely treatment costs, antibiotic costs, other drug costs, laboratory and radiology costs. The average total direct medical costs of patients who received ceftriaxone antibiotic therapy were more cost minimal than patients who received cefotaxime antibiotics, namely Rp.2,296,449 and Rp.2,464,470. The results of the difference test using independent t-test statistics show a p-value of 0.219 so that it can be interpreted that there is no significant difference.

Characteristics of Pulmonary Inflammation in Patients with Different Forms of Active Tuberculosis.

Targeted treatment of tuberculosis-associated lung damage requires an understanding of the precise mechanisms of immunopathology. A major obstacle to the longitudinal study of tuberculosis (TB) immunopathogenesis in humans is the lack of serial lung biopsies during disease progression and treatment, which could be used to characterize local immune pathways involved in tissue damage. Understanding of the immunobiology of lung tissue damage in tuberculosis has largely been based on animal models. Our study looked for signs of inflammation in TB patients' lung biopsies. Results were compared between a site of infection and relatively healthy tissue outside the site. The most significant differences in the expression of microRNAs (miRs) and cytokine/chemokines were observed between the non-decayed tuberculoma and the surrounding parenchyma. In addition, these parameters showed almost no differences between the cavitary wall and surrounding tissue. This is an indication that the inflammatory process is more prevalent in fibrotic cavitary tuberculosis (FCT). In FCT subjects, no difference was observed between the cavity wall and the parenchyma in the production of key inflammatory factors such as IL-6, IL-11, IL-17, and IFNγ. This is an indication that the limits of the inflammatory response are broader in FCT. The expression levels of miR-191, miR-193a, miR-222, miR-223, miR-18, miR-155, miR-376c, miR-26a, miR-150, and miR-124 were not significantly different between the cavernous wall and lung tissue in patients with FCT, further confirming the spread of inflammatory and destructive processes beyond the focus of infection.

Spatially resolved gene expression profiles of fibrosing interstitial lung diseases

Fibrosing interstitial lung diseases (ILDs) encompass a diverse range of scarring disorders that lead to progressive lung failure. Previous gene expression profiling studies focused on idiopathic pulmonary fibrosis (IPF) and bulk tissue samples. We employed digital spatial profiling to gain new insights into the spatial resolution of gene expression across distinct lung microenvironments (LMEs) in IPF, chronic hypersensitivity pneumonitis (CHP) and non-specific interstitial pneumonia (NSIP). We identified differentially expressed genes between LMEs within each condition, and across histologically similar regions between conditions. Uninvolved regions in IPF and CHP were distinct from normal controls, and displayed potential therapeutic targets. Hallmark LMEs of each condition retained distinct gene signatures, but these could not be reproduced in matched lung tissue samples. Based on these profiles and unsupervised clustering, we grouped previously unclassified ILD cases into NSIP or CHP. Overall, our work uniquely dissects gene expression profiles between LMEs within and across different types of fibrosing ILDs.

Hemin attenuates bleomycin-induced lung fibrosis in mice by regulating the TGF-β1/MAPK and AMPK/SIRT1/PGC-1α/HO-1/NF-κB pathways.

The objective of this study was to investigate the protective effect and potential mechanism of action of hemin on bleomycin-induced pulmonary fibrosis in mice. Male C57BL/6 mice were randomly divided into control, bleomycin and bleomycin + hemin groups. Mice in the bleomycin and bleomycin + hemin groups were injected intratracheally with bleomycin to establish the pulmonary fibrosis model. The bleomycin + hemin group mice were injected intraperitoneally with hemin starting 7 days before modeling until the end of Day 21 after modeling. Pathological changes in lung tissue were assessed by HE and Masson staining. Malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) levels were determined in lung tissue. Immunohistochemistry was performed to assess the expression of α-SMA and collagen I. The serum levels of IL-6 and TNF-α were measured via ELISA. Western blotting was used to determine the expression of TGF-β1, SIRT1, PGC-1α and HO-1 and the phosphorylation levels of p38, ERK1/2, JNK, AMPK and NF-κB p65 in lung tissue. Hemin significantly reduced lung indices, increased terminal body weight. It also significantly increased SOD and CAT activities; decreased MDA, IL-6 and TNF-α levels; reduced the levels of α-SMA and collagen I-positive cells; upregulated SIRT1, PGC-1α and HO-1 expression; promoted AMPK phosphorylation; and downregulated TGF-β1 expression and p38, ERK1/2, JNK and NF-κB p65 phosphorylation. Hemin might attenuate oxidative damage and inflammatory responses and reduces extracellular matrix deposition by regulating the expression and phosphorylation of proteins associated with the TGF-β1/MAPK and AMPK/SIRT1/PGC-1α/HO-1/NF-κB pathways, thereby alleviating bleomycin-induced pulmonary fibrosis.

Ginsenoside Rb1 targets to HO-1 to improve sepsis by inhibiting ferroptosis

Sepsis remains the leading cause of mortality among Intensive Care Unit (ICU) patients, with its pathogenesis and treatment not yet fully elucidated. Ferroptosis plays a critical role in sepsis, suggesting that ferroptosis-related genes may serve as potential therapeutic targets. This study aims to identify key ferroptosis-related genes in sepsis and explore targeted therapeutics. Through differential expression analysis of the GSE13940 and GSE26440 datasets, heme oxygenase-1 (HO-1) was identified as a hub gene associated with ferroptosis. Additionally, single-cell analysis of the GSE175453 dataset revealed a significant upregulation of HO-1 expression in monocyte lineages during sepsis. The cecal ligation and puncture (CLP) method was employed to induce sepsis in a mouse model, lung and intestinal tissues exhibited typical ferroptosis characteristics, with a significant increase in HO-1 expression. However, treatment with the HO-1 inhibitor zinc protoporphyrin (ZNPP) significantly ameliorated ferroptosis in CLP-induced lung and intestinal tissues, as well as in lipopolysaccharide (LPS)-induced THP-1 cells. Subsequently, molecular docking, surface plasmon resonance (SPR), and microscale thermophoresis (MST) experiments demonstrated that ginsenoside Rb1 specifically targets HO-1, identifying K18A as the key binding residue. Finally, experiments conducted both in vitro and in vivo verified that ginsenoside Rb1 significantly reduces HO-1 expression, inhibits ferroptosis in sepsis-induced lung, and intestinal tissues and THP-1 cells, and improves sepsis-induced pulmonary and intestinal damage. In conclusion, this study identifies HO-1 as a key ferroptosis target in sepsis and suggests ginsenoside Rb1 as a potential novel HO-1 inhibitor for the therapeutic approach of sepsis-induced organ dysfunction.

MPO interacts with hRSV particles, contributing to the virucidal effects of NETs against clinical and laboratory hRSV isolates

Human Respiratory syncytial virus (hRSV) mainly affects immunosuppressed patients requiring hospitalization. No specific treatment is financially accessible, and available vaccines do not cover all risk groups. During hRSV infection, there is a robust neutrophilic influx into the airways. hRSV-activated neutrophils release substantial neutrophil extracellular traps (NETs) in lung tissue, comprising DNA, histones, cytosolic, and granular proteins. NETs form mucus buildup in the lungs, compromising respiratory capacity and neutralizing viral particles. Understanding responsible NETs molecules requires improvement. We evaluated NETs interacting with hRSV particles and their contribution to anti-hRSV NET effects. Immunoblotting, immunoprecipitation, and peptide sequencing assays confirmed hRSV binding to a 50–75 kDa NET protein, Myeloperoxidase (MPO). MPO, a microbicide enzyme in NETs, interacts with hRSV, likely at F0 protein (site IV) on the viral surface. Additionally, MPO (32 μM) and NETs (0.4 μg/mL) reduced in vitro replication of clinical (hRSV A and B) and laboratory (Long) hRSV isolates by approximately 30 %, reversible by selective MPO inhibitor (PF-06281355; 48 μM). Thus, MPO contributes to virucidal NET effects on diverse hRSV strains, enhancing comprehension of NETs' role in infection and aiding treatment strategies for respiratory diseases.

Personalized in silico model for radiation-induced pulmonary fibrosis.

Radiation-induced pulmonary fibrosis (RIPF) is a severe late-stage complication of radiotherapy (RT) to the chest area, typically used in lung cancer treatment. This condition is characterized by the gradual and irreversible replacement of healthy lung tissue with fibrous scar tissue, leading to decreased lung function, reduced oxygen exchange and critical respiratory deficiencies. Currently, predicting and managing lung fibrosis post-RT remains challenging, with limited preventive and treatment options. Accurate prediction of fibrosis onset and progression is therefore clinically crucial. We present a personalized in silico model for pulmonary fibrosis that encompasses tumour regression, fibrosis development and lung tissue remodelling post-radiation. Our continuum-based model was developed using data from 12 RT-treated lung cancer patients and integrates computed tomography (CT) and dosimetry data to simulate the spatio-temporal evolution of fibrosis. We demonstrate the ability of the in silico model to capture the extent of fibrosis in the entire cohort with a less than 1% deviation from clinical observations, in addition to providing quantitative metrics of spatial similarity. These findings underscore the potential of the model to improve treatment planning and risk assessment, paving the way for more personalized and effective management of RIPF.

The preparation of casein-chitosan based naringin nanoparticles and their therapeutic application against Pseudomonas aeruginosa-induced pneumonia

Naringenin is a natural dihydro-flavonoid compound with various beneficial pharmacological activities. However, the extremely poor water solubility and bioavailability pose significant challenges for its application. In this study, chitosan-casein-naringenin nanoparticles (Cs-cas-Nar) were prepared to enhance naringenin's water solubility and bioavailability. Cs-cas-Nar had a particle size of 225.8 nm, a PDI of 0.155, a zeta potential of 27.2 mV, and an encapsulation efficiency of 87.02 %. FT-IR analysis indicated that casein encapsulated naringenin through hydrogen bonds and non-covalent interactions. Simulated gastrointestinal digestion and in vivo release results demonstrated that Cs-cas-Nar significantly improved naringenin's water solubility and bioavailability. The site-controlled and rapid release of Cs-cas-Nar provided higher drug concentration in the intestine and lung tissue. In in vivo experiments, Cs-cas-Nar provided a protective effect against Pseudomonas aeruginosa-induced pneumonia in mice. Cs-cas-Nar not only reduced weight loss and lung bacterial load of pneumonia mice, but also reduced the production of pro-inflammatory factors IL-1β and IL-6 by >50 % and restored the antioxidant capacity of the lungs. Through computer simulation technology and qPCR detection, we confirmed that Cs-cas-Nar can inhibit the assembly of P. aeruginosa flagella. These findings suggest that Cs-cas-Nar is a simple and easily manufactured formulation with potential application value in treating P. aeruginosa infection.

Etiologies and Comorbidities of Meningitis Deaths in Children Under 5 Years in High-Mortality Settings: Insights from the CHAMPS Network in the Post-Pneumococcal Vaccine Era

BackgroundThe role of meningitis in causing deaths and in children under 5 is unclear, especially since widespread use of vaccines to prevent common causes of meningitis. Child Health and Mortality Prevention Surveillance (CHAMPS) uses post-mortem minimally invasive tissue sampling (MITS) and ante-mortem data to explore death causes. We aimed to assess meningitis’s contribution to mortality and identify causative pathogens in children under 5 within CHAMPS Network sites. MethodIn this observational study, we analyzed deaths in live-born children <5 years of age that occurred between December 16, 2016, and December 31, 2023, in CHAMPS catchments in six sub-Saharan African countries (Ethiopia, Kenya, Mali, Mozambique, Sierra Leone, South Africa) and Bangladesh. MITS was conducted within 24–72hours of death, including blood and cerebrospinal fluid (CSF) culture, multi-organism targeted nucleic acid amplification tests on blood, CSF and lung tissue, and histopathology of lung, liver and brain. Expert panels at each site reviewed data to attribute causes of death following ICD-10 standards. ResultMeningitis was in the causal pathway for 7.0% (270/3857) of deaths; in 4.8% (13/270) meningitis was considered the underlying condition. Neonates accounted for 65.9% (178/270) and infants or children 34.1% (92/270). Among neonatal meningitis deaths, 55.6% (99/178) occurred ≥72hours post-hospital admission; and common pathogens were Acinetobacter baumannii (49.5%, 49/99; mainly from South Africa) and Klebsiella pneumoniae (40.4%, 40/99). Forty-four percent (79/178) of neonatal meningitis deaths were community-associated, primarily due to K. pneumoniae (35.4%, 28/79) and Escherichia coli (13.9%, 11/79). Among infant and child meningitis deaths, 43.5% (40/92) occurred ≥72hours post-admission; and common pathogens were K. pneumoniae (42.5%,17/40) and A. baumannii (17.5%, 7/40). Among community-associated meningitis deaths in infants and children (56.5%, 52/92), Streptococcus pneumoniae (34.6%, 18/52) and K. pneumoniae (19.2%, 10/52) were common pathogens. Pathogen prevalence varied by region. ConclusionOur study highlights meningitis as a significant contributor to under-5 mortality in low-middle-income countries. The prominent role of K. pneumoniae and A. baumannii, particularly in healthcare settings and specific regions, highlights the need for better infection control, targeted interventions, and more effective treatment strategies.

Yohimbine treatment improves pulmonary fibrosis by attenuating the inflammation and oxidative stress via modulating the MAPK pathway

Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disorder characterized by the accumulation of extracellular matrix and collagen, resulting in significant parenchymal scarring and respiratory failure that leads to mortality. Yohimbine (YBH) is an α-2 adrenergic receptor antagonist with anti-oxidant and anti-inflammatory properties. In the current study, we aimed to investigate the anti-inflammatory, anti-oxidant and anti-fibrotic activity of YBH against LPS/TGF-β-induced differentiation in BEAS-2B/LL29 cells and bleomycin (BLMN) induced pulmonary fibrosis model in rats. Network pharmacology, gene expression, Western-blot analysis, immune-cytochemistry/immunohistochemistry, lung function and histology techniques were used to assess the fibrotic marker expression/levels in cells or rat lung tissues. YBH treatment significantly attenuated the LPS-induced pro-inflammatory (identified through a network-pharmacology approach) and oxidative stress markers expression in lung epithelial cells. TGF-β stimulation significantly elevated the fibrotic cascade of markers and treatment with YBH attenuated these markers’ expression/levels. Intra-tracheal administration of BLMN caused a significant elevation of various inflammatory/oxidative stress and fibrotic markers expression in lung tissues and treatment with YBH significantly mitigated the same. Ashcroft score analysis revealed that BLMN exhibited severe distortion of the lungs, elevation of thickness of the alveolar walls and accumulation of collagen in tissues, further treatment with YBH significantly suppressed these events and improved the lung architecture. Lung functional parameters demonstrated that BLMN-induced stiffness and resistance were reduced considerably upon YBH treatment and restored lung function dose-dependently. Overall, this study reveals that YBH treatment significantly attenuated the BLMN-induced fibrosis by regulating the MAPK pathway and provided insightful information for progressing towards translational outcomes.

Dietary Eicosapentaenoic Acid Improves Ozone-Induced Pulmonary Inflammation in C57/BL6 Mice

Ambient concentrations of the air pollutant, ozone, are rising with increasing global temperatures. Ozone is known to increase incidence and exacerbation of chronic lung diseases, which will increase as ambient ozone levels rise. Studies have identified diet as a variable able to modulate the pulmonary health effects associated with ozone exposure. Eicosapentaenoic Acid (EPA) is an n-3 polyunsaturated fatty acid (PUFA) consumed through diet that lowers inflammation through conversion to oxylipins including Hydroxy Eicosapentaenoic Acids (HEPEs). However, the role of dietary EPA in ozone-induced pulmonary inflammation is unknown. Therefore, we hypothesize increasing dietary EPA will decrease ozone-induced pulmonary inflammation and injury through the production of HEPEs. To test this, male C57BL/6J mice were fed a purified control diet or EPA-supplemented diet for 4 weeks and then exposed to filtered air (FA) or 1 part per million ozone for 3 hours. 24 or 48 hours post exposure, bronchoalveolar lavage (BAL) fluid was collected to assess airspace inflammation/injury and lung tissue was collected for targeted Liquid Chromatography-Mass Spectrometry (LC-MS/MS) lipidomics. Following ozone exposure, EPA supplementation did not alter markers of lung injury, but decreased ozone-induced airspace neutrophilia. LC-MS/MS targeted lipidomics reveal dietary EPA supplementation increased pulmonary EPA-derived metabolites including 5-HEPE and 12-HEPE. Additionally, EPA supplementation decreased pulmonary levels of pro-inflammatory Arachidonic Acid (AA)-derived metabolites. To evaluate if dietary EPA reduces ozone-induced pulmonary inflammation through increased pulmonary HEPEs, C57BL/6J mice were administered 5- and 12-HEPEs systemically prior to filtered air or ozone exposure. Pretreatment with 5- and 12-HEPEs reduced ozone-driven increases in airspace macrophages. Together, these data indicate that an EPA-supplemented diet protects against ozone-induced airspace inflammation which is, in part, due to increasing pulmonary concentrations of 5- and 12-HEPEs. These findings suggest that dietary EPA and/or increasing EPA-derived metabolites in the lung can reduce ozone-driven incidences and exacerbations of chronic pulmonary diseases.

Improved Therapeutic Efficacy of Doxorubicin Chemotherapy With Cannabidiol in 4T1 Mice Breast Cancer Model.

High dose chemotherapy is one of the therapeutic strategies for breast cancer and doxorubicin (DOX) as a chemotherapy agent is widely used. DOX indication is limited due to its dose-depended cardiotoxicity. Recently, cannabidiol (CBD) shows antitumoral and cardioprotective effects, so we hypothesized that CBD administration with high-dose DOX chemotherapy can improve anticancer activity and reduce cardiotoxic side effects. Mice breast cancer model established by injecting 4T1 cell lines. One group was not injected by 4T1 cells as a not cancerous group and received normal saline (NS, 0.1 mL). In cancerous groups, first group was considered as cancerous control and received NS (0.1 mL); the second group received CBD (5 mg/kg, IP) on Days 1,7, and 14; in the third group DOX (5 mg/kg, IV) as CBD schedule was administrated; the fourth group treated with CBD 1 day before DOX injection as pretreatment, and the last group was treated with CBD and DOX at same time with previous doses and schedules. On Day 21, all mice were sacrificed, heart and lungs tissues were obtained and histological sections were isolated. SOD2, iNOS, MMP2, MMP9 were evaluated through western blot and TUNEL test preformed for breast tumor. Tumor size and weight significantly decreased in DOX, pretreatment CBD + DOX and CBD + DOX groups. Administration of CBD with DOX could not prevent weight loss. TUNEL test demonstrated the highest tumor cell apoptosis in pretreatment CBD + DOX and CBD + DOX. In lungs belonged to CBD + DOX, there was not any sign of metastasis. Cardiac histopathological examination of pretreatment CBD + DOX and CBD + DOX did not show any sign of congestion or inflammation. In CBD + DOX SOD2 increased, also iNOS, MMP2, and MMP9 decreased compared to DOX. This study demonstrated that simultaneous administration of CBD and DOX can increase antitumoral effect and reduce DOX cardiotoxicity. Nevertheless, CBD can induce cardiotoxicity as administrated alone.

Huaxian Formula Prevents the Progression of Radiation-Induced Pulmonary Fibrosis by Inhibiting the Pro-Fibrotic Effects of Macrophages

Ethnopharmacological relevanceThe Huaxian formula (HXF), a traditional Chinese medicine (TCM) remedy, specifically targets the pathological factors of "heat toxicity" and "phlegm stasis" induced by radiation in radiation-induced pulmonary fibrosis (RIPF). It works by clearing heat and invigorating the blood, addressing these key factors in the development of RIPF. Aim of the studyThe HXF has demonstrated potential in preventing RIPF, although its underlying mechanisms remain unclear. This study aims to investigate the efficacy, molecular targets, and mechanisms of action of HXF. Materials and methodsThe major constituents of the HXF were identified by ultra performance liquid chromatography and tandem mass spectrometry (UPLC-MS). C57BL/6j mice were divided into four groups: control (Ctrl), HXF alone (HXF), 17Gy-irradiation alone (IR), and irradiation plus HXF (IR + HXF). Lung damage and fibrosis were assessed by histopathological staining, and the flow cytometry and immunohistochemistry (IHC) were used to detect the macrophages phenotype of lung tissues in vivo at 16 weeks post-irradiation. Transcriptomic sequencing and bioinformatics analyses identified key genes modulated by HXF. In vitro assays included flow cytometry, western bolt, and quantitative PCR (qPCR) explored the impact of HXF on macrophage polarization and fibrotic activity, while co-culture experiments of the macrophage conditional medium and mouse embryo fibroblast NIH/3T3 investigated macrophage-fibroblast interactions. Results20 major constituents of HXF were identified. And the in vivo results revealed significant lung damage and fibrosis in the IR group, which were notably mitigated in the IR + HXF group. And HXF has been shown to significantly inhibit the infiltration of M2-type macrophages in lung tissues. Transcriptomic analysis identified differentially expressed genes (DEGs) such as Arg1, Mmp10, and Fgf23. Bioinformatics enrichment analysis indicated that these DEGs are involved in pathways related to the inhibition of extracellular matrix formation and inflammation. In vitro, HXF-containing serum reduced M2-type macrophage polarization and decreased the secretion of Arginase1 and TGFβ1. Conditioned medium from HXF-treated macrophages suppressed fibroblast activation. ConclusionHXF's preventive effects on RIPF involve multiple targets and mechanisms, including the modulation of Arg1, Mmp10, and Fgf23 expression. By inhibiting the pro-fibrotic capacity of macrophages, HXF suppresses fibroblast activation and collagen production, thereby alleviating lung fibrosis. These findings underscore the potential of HXF as a preventive strategy in managing RIPF.

Euphorbia helioscopia L. inhibits lung tumorigenesis through alleviating exhausted T cell induced by chronic inflammation

Ethnopharmacological RelevanceEuphorbia helioscopia L. (ZQ) is a very effective traditional Chinese medicine for treating pneumonia and lung cancer. However, the effects and mechanisms by which ZQ prevents lung tumorigenesis in the presence of chronic inflammation remain unexplored. AimTo examine the effects and mechanisms of ZQ in alleviating chronic inflammation-induced T cell exhaustion and inhibiting lung tumorigenesis. MethodsA mice model of lung tumorigenesis under chronic inflammation conditions was established by repeated administration of lipopolysaccharide (LPS) and exposure to the tobacco carcinogen nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). Mice were treated with ZQ ( 0.9, 1.8, and 3.6 g/kg/day) for 25 weeks. Lung pathology and tumor incidence were assessed, and inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF) and serum were measured. Additionally, the proportions of CD3+ T, CD4+ T, and CD8+ T cells and their inhibitory receptors expression were evaluated. In vitro, T cell exhaustion models were induced using inflammatory-conditioned media, followed by treatment with ZQ (0.5, 2, 8 μg/mL). T cell exhaustion markers and characteristics were analyzed, and molecular mechanisms were explored using RNA sequencing and Immunoblotting analysis. ResultsIn vivo, ZQ significantly reduced inflammatory infiltration and lung damage, tumor incidence, number, size, and lung and spleen indices in mice. It also markedly lowered the levels of pro-inflammatory cytokines and immunosuppressive cytokines in BALF and serum. Additionally, ZQ improved the proportions of CD3+ T, CD4+ T, and CD8+ T cells and significantly decreased the expression of inhibitory receptors on CD4+ T and CD8+ T cells in the lung tissues and spleen. In vitro, ZQ effectively alleviated T cell exhaustion induced by the inflammatory environment, marked by reduced expression of inhibitory receptors, increased cytokine secretion, improved proliferation, and enhanced tumoricidal activity. RNA sequencing revealed that ZQ significantly downregulated the JAK-STAT signaling and upregulated stemness-associated pathways. Immunoblotting results indicated that treatment with ZQ markedly reduced the phosphorylation of Signal transducer and activator of transcription 3 (STAT3) and increased the expression of T cell factor -1/7 (TCF1/7). ConclusionZQ inhibits lung tumorigenesis in LPS/NNK-treated mice through alleviating exhausted T cells induced by chronic inflammation, which is attributed to the suppression of STAT3 activation and the maintenance of stemness characteristics in T cells. These findings provide experimental evidence for the potential use of ZQ in preventing and treating lung tumourigenesis in patients with chronic inflammation and the clinical management of lung cancer patients with concomitant chronic inflammation.

Exploring an indirect quantification strategy for PFOA in rat tissues via efficient degradation and fluorescence spectroscopy

Perfluorooctanoic acid (PFOA) has aroused global attention due to its persistence, biotoxicity and bioaccumulative properties. Monitoring PFOA levels in biological samples could give significant insights into its toxicity mechanism and health risk evaluation. Herein, to explore the distribution of PFOA in biological tissues, an indirect quantitation strategy for PFOA in biological samples was proposed based on fluorescence spectroscopy. F- ion was produced through highly efficient degradation of PFOA in polar aprotic solvent, and then F- ion was detected with highly selective F- ion fluorescent probe, which enabled the indirect quantitative detection of PFOA in biological samples. The sensitivity for PFOA was exhibited with limit of detection of 3.55 μM. The method’s accuracy and precision were validated by spiked biological sample experiment, with recovery rate ranging from 93.2 % to 105.5 % and relative standard deviation below 9.2 %. The content of PFOA was successfully detected in rats livers, kidneys and lungs tissues. Density function theory was employed to explore defluorination pathway, indicating that heating, alkaline and water conditions played a crucial role in the degradation of PFOA to produce F- ion. Thus, this novel strategy is suitable for quantitative analysis of PFOA in biological samples with high selectivity, sensitivity, recovery ratio, and simple sample preparation, which also provides a strategy for evaluating the levels of per- and polyfluoroalkyl substances in biological samples.

Endoplasmic Reticulum Oxidative Stress Promotes Glutathione-Dependent Oxidation of Collagen-1A1 and Promotes Lung Fibroblast Activation.

Changes in the oxidative (redox) environment accompany idiopathic pulmonary fibrosis (IPF). S-glutathionylation of reactive protein cysteines is a post-translational event that transduces oxidant signals into biological responses. We recently demonstrated that increases in S-glutathionylation promote pulmonary fibrosis, which was mitigated by the deglutathionylating enzyme glutaredoxin (GLRX). However, the protein targets of S-glutathionylation that promote fibrogenesis remain unknown. In the present study we addressed whether the extracellular matrix is a target for S-glutathionylation. We discovered increases in COL1A1 (collagen 1A1) S-glutathionylation (COL1A1-SSG) in lung tissues from subjects with IPF compared with control subjects in association with increases in ERO1A (endoplasmic reticulum [ER] oxidoreductin 1) and enhanced oxidation of ER-localized PRDX4 (peroxiredoxin 4), reflecting an increased oxidative environment of the ER. Human lung fibroblasts exposed to TGFB1 (transforming growth factor-β1) show increased secretion of COL1A1-SSG. Pharmacologic inhibition of ERO1A diminished the oxidation of PRDX4, attenuated COL1A1-SSG and total COL1A1 concentrations, and dampened fibroblast activation. Absence of Glrx enhanced COL1A1-SSG and overall COL1A1 secretion and promoted the activation of mechanosensing pathways. Remarkably, COL1A1-SSG resulted in marked resistance to collagenase degradation. Compared with COL1, lung fibroblasts plated on COL1-SSG proliferated more rapidly and increased the expression of genes encoding extracellular matrix crosslinking enzymes and genes linked to mechanosensing pathways. Overall, these findings suggest that glutathione-dependent oxidation of COL1A1 occurs in settings of IPF in association with enhanced ER oxidative stress and may promote fibrotic remodeling because of increased resistance to collagenase-mediated degradation and fibroblast activation.

The adequacy of tissue obtained by percutaneous needle autopsy in comparison to conventional autopsy for lung and liver tissues

The adequacy of tissue obtained by percutaneous needle autopsy in comparison to conventional autopsy for lung and liver tissues

Tailoring the mechanical properties of macro-porous PVA hydrogels for biomedical applications

Polyvinyl alcohol (PVA) is a biocompatible biopolymer with superior dimensional and mechanical stability when compared to naturally available biomaterials such as collagen and gelatin. Furthermore, PVA in hydrogel form behaves non-linearly during mechanical loading, generating a response like soft biological tissues. Generally, PVA hydrogels are fabricated using freeze-thaw cycles (FTCs) and changing the number of FTCs gives control over its mechanical properties. Porosity of the hydrogel is another important factor which determines its mechanical properties and is also evident in biological soft tissues. Incorporating macro-pores in PVA hydrogels substantially reduces the stiffness of the material and can mimic some porous tissues such as lung, liver, bone marrow, kidneys, urethra and penile tissues (corpus cavernosa). Within this study, we developed macro-porous PVA hydrogels using the freeze-thaw process followed by particulate leaching of sacrificial 3D-printed and milled PVA (m-PVA) filler particles. This fabrication method enables control over the porosity in macro-porous PVA hydrogels, which is crucial not only for tuning mechanical properties but also for mimicking the structure of spongy tissues, such as liver tissue and corpus cavernosum in the penile tissue, for example. We investigated the level of porosity in the specimen using optical microscopy to understand the distribution of the pores and the pore size. The tunability of the mechanical properties of PVA hydrogels is, a key finding of this study and is achieved using three factors: (i) weight percentage of sacrificial fillers, (ii) number of FTCs and (iii) concentration of PVA. These macro-porous PVA specimens have wide ranging biomedical applications as biological soft tissue analogues, or tissue engineering scaffolds, where the PVA hydrogel can be tuned to match the mechanical properties of these soft biological tissues.

Novel therapeutic effects of rifaximin in combination with methylprednisolone for LPS-induced ‎oxidative stress and inflammation in mice‎: ‎an in vivo study

Cytokine-releasing syndrome (CRS) is a special form of ‎‎systemic inflammatory response syndrome provoked by ‎factors ‎like viral infections and certain immunomodulatory drugs.‎ To elucidate the potential ‎role of rifaximin (RIF) and its combination with methylprednisolone (MP) against the development and ‎progression of CRS in ‎mice.‎ This experiment consists of two parts: protective and therapeutic interventions. The protective experiment: in the induction group, mice received an intraperitoneal injection (IP) of 5mg/kg lipopolysaccharide (LPS) without intervention. The other group received various drugs before the induction by three days, then observed for an additional two days (50mg/kg MP, 50mg/kg RIF, and a combination of 25mg/kg RIF with 25mg/kg MP. The second part of the study involves the therapeutic potential; all groups received similar doses of drugs to that received in the prevention groups, except LPS induction was given first, and after one hour, the mice received daily doses of the drugs for five days. At the end of the experiment, blood and tissue samples were obtained. Mice treated with RIF and its combination with MP showed improved serum TNF-α, IL-6, IL-8, IL-1β, INF-γ, MDA, and GSH in both prevention and therapeutic groups. Histopathologically, mice treated with rifaximin and its combination with MP ameliorates the tissue damage in both lung and liver tissues following LPS induction. In conclusion, rifaximin showed protective and therapeutic effects in LPS-induced cytokine storms in mice through anti-inflammatory and antioxidant mechanisms, and its combination with methylprednisolone showed additive/ synergistic action.