Related Signaling Research Articles

Acid-Cleavable Guanosine Triphosphate-Photoaffinity Probe for Global Profiling of Guanosine Triphosphate-Binding Proteins and Their Active Sites.

Guanosine triphosphate (GTP)-binding proteins function as molecular switches in cell signaling, playing critical roles in various biological pathways. Their dysregulation is associated with the causes and progression of many diseases. Systematic analysis of GTP-binding proteins would facilitate studies of related signaling pathways and drugs. Previously reported acyl-phosphate GTP-affinity probes, which react with and label lysine residues near GTP-binding pockets, have proven efficient in identifying labeling sites but suffer from poor stability due to their high reactivity. We report here new GTP-photoaffinity probes that employ a UV-triggered photoreactive group for covalent labeling of proteins, greatly improving probe stability. The inclusion of a terminal alkyne group allows labeled proteins to be tagged either with a fluorophore for fluorescence analysis or with a biotin group to enrich for LC-mass spectrometry (MS)/MS analysis. We further designed a GTP-N probe featuring an acid-cleavable P-N bond. The P-N bond enabled the release of GTP from labeling sites upon incubation under acidic conditions after labeling and enrichment, which reduced protein-modification mass shift and facilitated MS-based modification-site identification. This new method demonstrates good potential for identifying new GTP-binding proteins and systematically analyzing GTP-binding sites. These novel GTP-photoaffinity probes could be further applied in studying related biochemical mechanisms and in evaluating GTPase inhibitors.

Structure-function relation of cytokinins determines their differential efficiency in mediating tobacco resistance against Pseudomonas syringae.

The classic plant growth-promoting phytohormone cytokinin has been identified and established as a mediator of pathogen resistance in different plant species. However, the resistance effect of structurally different cytokinins appears to vary and may regulate diverse mechanisms to establish resistance. Hence, we comparatively analysed the impact of six different adenine- and phenylurea-type cytokinins on the well-established pathosystem Nicotiana tabacum-Pseudomonas syringae. The efficiency of resistance effects was evaluated based on impacts on the host plant defence response by scoring infection symptoms and the direct impact on the pathogen by assessment of proliferation in planta. To identify common and cytokinin-specific components involved in resistance effects, transcriptome profiling and targeted metabolomics were conducted in leaves treated with the different cytokinins. We observed clearly different potentials of the tested cytokinins in either suppressing infection symptoms or pathogen proliferation. Gene regulation and metabolite analyses revealed cytokinin-type specific impacts on defence components, such as salicylic acid and related signalling, expression of PR proteins, and regulation of specialised metabolism. Cytokinins also strongly affected plant cell physiological parameters, such as a remarkable decrease in amino acid pools. Hence, this study provides comparative information on the efficiency of diverse cytokinins in mediating resistance in one well-studied pathosystem and insights into the specific regulation of resistance effects mediated by different cytokinin molecules. This is particularly relevant for studies on the function of cytokinins or other phytohormones and compounds interacting with cytokinin activities in the context of pathogen infections and other stress scenarios, considering the diverse cytokinins present in plants.

Isobavachin attenuates FcεRI-mediated inflammatory allergic responses by regulating SHP-1-dependent Fyn/Lyn/Syk/Lck signaling

Isobavachin, isolated from Psoralea corylifolia L. exhibits therapeutic potential for osteoporosis or skin disease. Here, we evaluated the pharmacological effects of isobavachin on IgE-dependent inflammatory allergic reactions, as well as the underlying mechanisms, in bone marrow-derived mast cells and a mouse model of passive cutaneous anaphylaxis (PCA). Isobavachin reduced IgE/Ag-stimulated degranulation, eicosanoid (leukotriene C4 and prostaglandin D2) generation, and release of pro-inflammatory cytokines (tumor necrosis factor-α (TNF-α) and interleukin (IL)-6). Mechanistic studies revealed that isobavachin suppressed activation of Fyn, Lyn, spleen tyrosine kinase (Syk), and lymphocyte-specific-protein-kinase (Lck), receptor-proximal tyrosine kinases that initiate and play a central role in FcɛRI-mediated mast cell activation, as well as their common downstream signaling molecules including linker for activation of T cells, phospholipase Cγ1, AKT, mitogen-activated protein kinases (MAPKs), and intracellular Ca2+. Additionally, isobavachin increased phosphorylation of Src homology region 2 domain-containing phosphatase-1 (SHP-1), thereby strengthening its interaction with Syk and Lck as well as Fyn and Lyn, resulting in de-phosphorylation of these proximal tyrosine kinases. Genetic knockdown of SHP-1 reversed the inhibitory effects of isobavachin on mast cell activation, as well as the related signaling pathways, indicating that the inhibitory effects of isobavachin are mediated by negative regulation of SHP-1-dependent Fyn, Lyn, Syk and Lck. The anti-inflammatory properties of isobavachin were also examined in macrophages. Isobavachin suppressed production of lipopolysaccharide-stimulated production of pro-inflammatory cytokines and nitric oxide. Furthermore, oral administration of isobavachin attenuated mast cell-mediated PCA reactions in mice. These results suggest that isobavachin is a potential treatment for mast cell-mediated allergic inflammatory diseases.

Abstract TP338: Functional recovery during the chronic phase after cerebral infarction via CSF-derived microRNA

(Background): Stroke has become a leading cause of mortality and disability worldwide. However, except rehabilitation, there is no effective therapy during chronic phase after ischemic stroke. Interestingly, it has been reported that young cerebrospinal fluid (CSF) restores oligodendrogenesis and memory in aged mice. However, if specific components in CSF could contribute to functional recovery during chronic phase of ischemic stroke remains unclear. We tried to distinguish a specific microRNA in CSF and identify its potential therapeutic functions. (Method): Rats were subjected to permanent left middle cerebral artery occlusion (pMCAO). Brains and CSF were collected at 3, 7, 14, 28 days after pMCAO. MicroRNA in CSF was evaluated by microRNA Array. A specific microRNA was selected based on heatmap and volcano plot. IPA analysis was used to distinguish possibly related signaling pathways. Mimic/ inhibitor of this microRNA was applied respectively to primary cultured astrocytes and neurons soon after 3 hours’ oxygen glucose deprivation (OGD). Associated proteins were tested through western blot at 96 hours after OGD. In vivo, time course of pStat3 was assessed in peri-infarct cortical area. At 7 days after pMCAO, mimic of this microRNA was applied to CSF through Cisterna Magna. behavioral tests were assessed at 7, 14, 21, 28 days after pMCAO. At 28 days after pMCAO, brains were harvested for ischemia volume calculation and immunofluorescence. (Result): miR-204-5p was significantly upregulated in CSF at 7 days after pMCAO and selected as target microRNA. According to IPA analysis, miR-204-5p might be associated with Stat3 signaling pathway. Based on results of astrocyte culture, GFAP was significantly decreased by applying miR-204-5p mimic. Regarding neuron culture, pStat3, MAP2 showed a tendency to increase by applying miR-204-5p mimic. In vivo, pStat3 significantly increased at 3 days and began to decrease from 7 days after pMCAO. miR-204-5p mimic group showed a tendency of ameliorated functional dysfunction, smaller ischemia volume at 28 days after pMCAO. In peri-infarct area, compared with non-treatment group, GFAP was significantly decreased, and pStat3 showed tendency to increase in miR-204-5p mimic group. (Conclusion): Applying miR-204-5p mimic into CSF contributed to functional recovery via activating Stat3 signalling pathway and reducing astrocytosis, which might be a candidate of exosome or liposome preparation for future clinical trials.

Expression Profile of Thymidine Kinase Genes in Cervical Squamous Cell Carcinoma Confirmed by Various Detection Methods.

Thymidine kinases (TKs) are key enzymes involved in DNA synthesis and repair, with alterations in their expression associated with various cancers. Thymidine kinase 1 (TK1) and TK2 are cytosolic enzyme proteins that catalyze the addition of a gamma-phosphate group to thymidine. The existing literature on TK1 in cervical squamous cell carcinoma (CESC) fails to address the clinical role of TK1 overexpression and its possible molecular mechanism in CESC. The clinical significance of TK2 in CESC is also unknown. The objective was to explore the differential expression, clinical significance, and molecular mechanisms of TK1 and TK2 in CESC. The researchers collected global high-throughput data, extracted the expression levels of TK1 and TK2, and calculated the integrated standardized mean difference (SMD) and summarized receiver's operating characteristics (sROC) of TK1 or TK2 mRNA to investigate the expression profiles of TK genes fully and objectively in 918 CESC tissues and 360 control tissues. In-house tissue microarrays for immunohistochemical testing were used to verify the protein level of TK1 in 62 CESC tissues and control tissues. The growth effect of TK1 and TK2 in CESC cell lines was assessed using Chronos dependency scores derived from CRISPR knockout screen in the Achilles project. We also analyzed the potential mechanism of TK genes by studying the relationship between TK gene expression and immune infiltration, gene alternations as well as the related signal pathways. The various detection methods employed all confirmed that the TK1 expression is upregulated and TK2 is downregulated in CESC tissues (SMD: 2.44, 95% confidence interval (CI): 1.36 - 3.51, area under curve (AUC): 0.88, 95% CI: 0.85 - 0.90; SMD: -0.69, 95% CI: -1.25 to -0.14, AUC: 0.75, 95% CI: 0.71 - 0.78). Inhibition of TK1 expression by CRISPR knockout had negative influence on the biological functions of 11 CESC cell lines. The expression of TK2 was negatively correlated with the malignant progression of CESC. Expression of TK genes showed significant association with the immune infiltration of macrophages, CD4+ T cells, and neutrophils. Genes related with TK1 or TK2 were involved in pathways related to DNA replication, proteasome, and homologous recombination. Clinically, these findings suggest that the differential expression of TK1 and TK2 could serve as potential biomarkers, as well as therapeutic targets for personalized treatment strategies in CESC patients.

A comprehensive analysis of the pyruvate kinase M1/2 (PKM) in human cancer.

Pyruvate Kinase Muscle Isozyme (PKM), as a member of the pyruvate kinase, is a key enzyme in glycolysis. Numerous tumors have demonstrated its oncogenic properties. There is, however, no pan-carcinogenic analysis for PKM. A thorough analysis of PKM across various types of cancer was carried out using bioinformatics resources like The National Cancer Institute's Clinical Proteomic Tumor Analysis Consortium (CPTAC) and The Cancer Genome Atlas (TCGA) database. This study involved analyzing the role of PKM in 33 various types of cancers, along with investigating gene expressions, survival rates, clinical importance, genetic changes, immune system presence, and related signaling pathways. Furthermore, we evaluated the effects of PKM knockdown on human colon carcinoma, and glioblastoma cell lines by in vitro experimentation. In most tumors, PKM expression was markedly increased and was associated with unfavorable overall survival (OS) in certain individuals. In addition, infiltration of macrophages was associated with PKM expression in various tumors. PKM was linked to glycolysis/gluconeogenesis, HIF-1 signaling, carbon metabolism, and NADPH regeneration in a mechanistic manner. Additionally, cell experiments showed that the knockdown of PKM could reduce the proliferation and migration abilities while promoting the apoptosis of Caco-2, and U-87 MG cells. PKM controls immune cell infiltration, impacts patient outcomes in various types of cancer, and plays an essential role in proliferation and migration in some tumor cells by affecting glycometabolism. The PKM molecule may serve as a potential prognostic biomarker and therapeutic target for human cancers.

Molecular and cellular mechanisms of PDAC progression based on RETN-CAP1-mediated macrophage-fibroblast crosstalk: Action of ITGB5 and ITGB1 recombinant proteins.

Pancreatic ductal adenocarcinoma (PDAC) has a very poor prognosis, and the main objective of this study was to reveal the specific mechanism of action of TN-CAP1-mediated macrophage-fibroblast crossinulation in the progression of PDAC, and to evaluate the function and potential therapeutic value of ITGB5 and ITGB1 recombinant proteins in this process. The expression of TN-CAP1 in tumor tissues of PDAC patients was analyzed by immunohistochemistry and compared with normal pancreatic tissues. The co-culture system of macrophages and fibroblasts was constructed using in vitro cell culture model. The intercellular interactions and their effects on the proliferation, migration and invasion of tumor cells were observed by adding or knocking down ITGB5 and ITGB1 proteins. Western blot and RT-PCR were also used to detect the expression changes of related signaling pathway proteins and mRNA, which verified the effects of ITGB5 and ITGB1 recombinant proteins on tumor growth and metastasis in vivo. In vitro experiments showed that the addition of ITGB5 and ITGB1 recombinant proteins significantly enhanced the interaction between macrophages and fibroblasts, and promoted the proliferation and migration of tumor cells. Specifically, ITGB5 and ITGB1 recombinant proteins promote tumor cell aggressiveness by activating the FAK/PI3K/AKT signaling pathway.

CTSL Promotes Autophagy in Laryngeal Cancer Through the IL6-JAK-STAT3 Signalling Pathway.

Cathepsin L (CTSL) is an important oncogene. However, its mechanism of action in laryngeal cancer is still unclear. This study aims to explore the role of CTSL in laryngeal cancer and its clinical significance. Conducting bioinformatics analysis utilising the Cancer Genome Atlas (TCGA) database and the Gene Expression Omnibus (GEO) database. Performing CCK8 analysis, Western blotting, qRT-PCR, wound healing assay and transwell invasion assay. Additionally, conducting immunoprecipitation experiments and immunohistochemical staining to investigate the impact of CTSL on cell proliferation, autophagy and related signalling pathways. We observed a significant upregulation of CTSL in laryngeal cancer tissues, and its elevated levels are indicative of poor prognosis in laryngeal cancer patients. The proliferation of laryngeal cancer cells relies on the expression of CTSL, with overexpression of this gene enhancing the proliferative capacity of these cells. Concurrently, CTSL is closely associated with the autophagic levels in laryngeal cancer cells. During the autophagic process mediated by CTSL, the IL6-JAK-STAT3 signalling pathway is activated, suggesting that CTSL promotes autophagy through the IL6-JAK-STAT3 pathway. Considering the correlation between CTSL and autophagy, we developed and validated a multi-gene signature. The risk score derived from this signature demonstrates significant potential in predicting various aspects. We found that CTSL upregulates autophagy in laryngeal cancer cells by activating the IL6-JAK-STAT3 signalling pathway. By taking into account the autophagy-regulating role of CTSL, the clinical predictive ability of CTSL in HNSC can be enhanced.

Enhanced underwater three-dimensional imaging using acoustic orbital angular momentum waves and mode matching beamforming.

Improving the underwater three-dimensional imaging resolution of a sonar system is of great significance to achieving high-precision ocean exploration results. Actually, improving the resolution can be considered from the perspective of information acquisition. The acoustic orbital angular momentum (AOAM) wave has a modal dimension and can carry more target information. However, there are few studies on the application of AOAM waves for underwater three-dimensional imaging. This paper establishes the related signal models of the AOAM wave in underwater imaging and examines the sound field characteristics from both simulation and underwater real test data. A method called mode matching beamforming (MMBF) is proposed, and a composite structure of uniform circular array (UCA) plus spiral array is combined to study the imaging performance of AOAM waves and plane waves. The simulation results indicate that, compared with the plane wave, the sidelobe in the AOAM wave imaging beam pattern reaches -22.43 and -29.10 dB in the azimuth and elevation angles, respectively, and the mainlobe widths are reduced by 1.70° and 0.40° in both directions. Finally, this paper uses the MMBF method to process the echo data of underwater real object and realizes the imaging of the corner reflector.

Hepatotoxicity of Phytolacca acinosa Roxb mediated by phytolaccagenin via ferroptosis/PPAR/P53/arachidonic acid metabolism.

The traditional Chinese medicine Phytolacca acinosa Roxb (PAR), known as Shanglu, possesses recognized therapeutic benefits against many diseases. PAR is also hepatotoxic, making it a major public health problem. However, the specific toxic substances and molecular mechanisms of PAR remain unclear. Therefore, appropriate animal models and methods are essential to confirm the toxic components and related mechanisms of PAR. L-02 cells and zebrafish larvae at 4 days post-fertilization (4 dpf) were used as models and treated with various concentrations of phytolaccagenin (Phy), esculentoside A (EsA), and esculentoside H (EsH). The hepatotoxicity of three samples was assessed based on liver phenotype, pathological assessments, and biochemical index in zebrafish and proliferative activity, apoptosis level, and biochemical index in L02 cells. The transcriptomic technique was used to explore the related signaling pathways and potential mechanisms in vitro and in zebrafish , and the findings were validated by RT-PCR. The results of acute toxicity tests indicated that Phy exhibited substantially more severe hepatotoxicity than EsA, while EsH did not lead to any obvious toxic effects. Especially, under sublethal exposure (<LC10), both Phy and EsA induced similar liver damage in zebrafish and L-02 cells, increasing mortality, disrupting morphology, enhancing apoptosis, altering liver enzyme levels, and leading to significant structural changes in cells and zebrafish. Multiomics analysis of 605 genes in L-02 cells and 780 genes in zebrafish showed that exposure to Phy significantly altered gene expression in various biological processes. Further enrichment analysis demonstrated that Phy predominantly affects the P53\apoptosis\cell cycle arrest, ferroptosis, PPAR signaling, and arachidonic acid metabolism, leading to notable cellular damage. This study identified Phy as a key hepatotoxic component of PAR. Furthermore, using transcriptomic techniques, we preliminarily investigated the hepatotoxic mechanisms of Phy in vitro and in vivo. The results of the present study showed that Phy affects several signaling pathways, including those involved in lipid metabolism, oxidative stress, and apoptosis, finally leading to hepatotoxicity. These findings provide invaluable insights into the safe use of PAR in clinical settings.

Identification of gene expression change associated with isoflurane anesthesia and neurocognitive disorders using bioinformatics methods.

Isoflurane, a commonly used anesthetic, has been linked to neurocognitive dysfunction (NCD). However, the precise relationship between isoflurane anesthesia and NCD remains unclear. Datasets related to isoflurane anesthesia were obtained from a public database. The hub genes of isoflurane anesthesia were selected using logistic regression with the least absolute shrinkage and selection operator (LASSO). Human neuroblastoma cell line SH-SY5Y was induced to differentiate into terminal neuron-like cells. The signaling pathways involved in isoflurane anesthesia and NCD were subjected to analysis, resulting in the identification of 35 signaling pathways, including the PI3K/AKT pathway, the AGE-RAGE signaling pathway and the apelin signaling pathway, which were found to be associated with both NCD and isoflurane anesthesia. A total of nine cross-expressed genes (Nfe2l2, Fgf2, Edn1, Spp1, Hmox1, Picalm, Gnb5, Eif2s1 and Gls) were identified between isoflurane anesthesia and NCD. The LASSO logistic regression model was employed to identify three hub genes (Fgf2, Gnb5, Spp1). The expression of these three hub genes was validated using other expression datasets. In human neuron-like cells, isoflurane also significantly affected the expression of three corresponding human homologous genes. The expression trends of these three genes in human cells were consistent with the expression in rat brains after isoflurane treatment, providing further evidence for the rationality of the three hub genes. The present study revealed key candidate genes and related functional signaling pathways through bioinformatics analysis and cell experiments, which may serve as the basis for isoflurane-related NCD.

Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network

Neuromuscular diseases usually manifest as abnormalities involving motor neurons, neuromuscular junctions, and skeletal muscle (SkM) in postnatal stage. Present in vitro models of neuromuscular interactions require a long time and lack neuroglia involvement. Our study aimed to construct rodent bioengineered spinal cord neural network-skeletal muscle (NN-SkM) assembloids to elucidate the interactions between spinal cord neural stem cells (SC-NSCs) and SkM cells and their biological effects on the development and maturation of postnatal spinal cord motor neural circuits. After coculture with SkM cells, SC-NSCs developed into neural networks (NNs) and exhibited a high proportion of glutamatergic and cholinergic neurons, low proportion of neuroglia and gamma-aminobutyric acidergic neurons, and increased expression of synaptic markers. In NN-SkM assembloids, the acetylcholine receptors of SkM cells were upregulated, generating neuromuscular junction-like structures with NNs. The amplitude and frequency of SkM cell contraction in NN-SkM assembloids were increased by optogenetic and glutamate stimulation and blocked by tetrodotoxin and dizocilpine, respectively, confirming the existence of multisynaptic motor NNs. The coculture process involves the secretion of neurotrophin-3 and insulin growth factor-1 by SkM cells, which activate the related ERK-MAPK and PI3K-AKT signaling pathways in NNs. Inhibition of the ERK-MAPK and PI3K-AKT pathways significantly reduces neuronal differentiation and synaptic maturation of neural cells in NN-SkM assembloids, while also decreasing acetylcholine receptor formation on SkM cells. In brief, NN-SkM assembloids simulate the composition of spinal cord motor NNs and respond to motor regulatory signals, providing an in vitro model for studying postnatal development and maturation of spinal cord motor NNs.

Early-onset sleep alterations found in patients with amyotrophic lateral sclerosis are ameliorated by orexin antagonist in mouse models.

Sleep alterations have been described in several neurodegenerative diseases yet are currently poorly characterized in amyotrophic lateral sclerosis (ALS). This study investigates sleep macroarchitecture and related hypothalamic signaling disruptions in ALS. Using polysomnography, we found that both patients with ALS as well as asymptomatic C9ORF72 and SOD1 mutation carriers exhibited increased wakefulness and reduced non-rapid eye movement sleep. Increased wakefulness correlated with diminished cognitive performance in both clinical cohorts. Similar changes in sleep macroarchitecture were observed in three ALS mouse models (Sod1G86R, FusΔNLS/+, and TDP43Q331K). A single oral administration of a dual-orexin receptor antagonist or intracerebroventricular delivery of melanin-concentrating hormone (MCH) through an osmotic pump over 15 days partially normalized sleep patterns in mouse models. MCH treatment did not extend the survival of Sod1G86R mice but did decrease the loss of lumbar motor neurons. These findings suggest MCH and orexin signaling as potential targets to treat sleep alterations that arise in early stages of the disease.

3D imaging of underwater scanning photon counting lidar based on multiscale spatio-temporal resolution

This manuscript presents an underwater scanning photon counting lidar system specifically designed for the 3D imaging of underwater targets. A multiscale spatio-temporal resolution method is proposed to enhance the accuracy and resolution of 3D scanning imaging. Signal extraction is achieved through macro pulse accumulation number and macro time resolution, based on the spatio-temporal correlation constrained by relative signal intensity difference factor. Subsequently processing focuses exclusively on the photon-counting events extracted in the previous step. By employing micro pulse accumulation number and micro time resolution, each pixel is expanded into multiple pixels, thereby improving timing precision. This not only enhances imaging accuracy but also enables the detection of more detailed information about the target. The reconstructed images of the resolution plate located approximately 3.5 meters away demonstrate that both the imaging accuracy and resolution are within 10.0 mm. In addition, high-performance 3D reconstruction of a coral model located approximately 4 meters away with complex surface shape is also realized, where each branch of the coral can be distinctly identified. It is verified that the developed lidar system has the ability of high-performance 3D imaging for underwater targets. This lidar system will play an important role in the field of fine mapping of seabed topography and underwater target detection and recognition.

The cellular response of lipopolysaccharide-induced inflammation in keratoconus human corneal fibroblasts to RB-PDT: Insights into cytokines, chemokines and related signaling pathways.

Rose Bengal Photodynamic Therapy (RB-PDT) offers dual therapeutic benefits by enhancing corneal stiffness and providing antibacterial activity, presenting significant potential for patients with keratoconus complicated by keratitis. Our purpose was to assess the effect of rose bengal photodynamic therapy (RB-PDT) on the expression of pro-inflammatory cytokines and chemokines, as well as on extracellular matrix (ECM)-related molecules, in lipopolysaccharide (LPS)-induced inflammation of keratoconus human corneal fibroblasts (KC-HCFs). Additionally, the involvement of the mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways which are downstream of the Toll-like receptor 4 (TLR4) pathway were examined. KC-HCFs were stimulated with varying concentrations of LPS (0-10 μg/ml), which was followed by RB-PDT. The expression levels of interleukin-1β (IL-1β), IL-6, IL-8, interferon alpha 2 (IFNA2), IFNB1, intercellular adhesion molecule 1 (ICAM-1), chemokine (C-C motif) ligand 4 (CCL-4), collagen I, collagen V, lysyl oxidase (LOX), transforming growth factor β 1(TGF-β1) were measured using qPCR, ELISA, or western blot. The activation of the NF-κB and MAPK pathways was assessed using qPCR and western blot. In LPS-induced inflammation of KC-HCFs, the expression of IL-6 was further amplified by the treatment with RB-PDT (p = 0.001). However, the activation of the MAPK and NF-κB pathways did not increase following RB-PDT. Additionally, RB-PDT reduced the transcription of collagen I and collagen V (p≤0.03), while the transcription of LOX and TGF-β1 secretion remained unchanged in KC-HCFs exposed to LPS. In LPS-induced inflammation of KC-HCFs treated with RB-PDT, despite the increased expression of pro-inflammatory cytokines, the activation of the TLR4 signaling pathways is lacking. RB-PDT may have no adverse effects on corneal scar formation of keratoconus corneas in the short term.

Bacopa monnieri Extract Diminish Hypoxia-Induced Anxiety by Regulating HIF-1α Signaling and Enhancing the Antioxidant Defense System in Hippocampus.

Hypoxia is a significant stressor, and stabilized hypoxia-inducible factor-1α (HIF-1α) regulates the expression of numerous genes, leading to various biochemical, molecular, physiological and genomic changes. The body's oxygen-sensing system activates gene expression to protect brain tissues from hypoxia. Gamma-aminobutyric acid, an inhibitory neurotransmitter, regulates brain excitability during hypoxia through the activation of HIF-1 α. Herbal medicines have been widely used for managing various toxicological effects and disorders including hypoxia; however, the data on safety, efficacy and the molecular mechanisms that increase vulnerability or lethality against hypoxia are still lacking and urgently need to be investigated. The Current study aims to investigate how Bacopa monnieri extract (BME), specially CDRI-08 affects the hippocampus of mice subjected to conditions that simulate hypoxia. The pre and co-treatment of mice involved administrating BME (200mg/kg BW) for 14days, followed by exposure to CoCl2 (40mg/kg BW). BME decreased the levels of reactive oxygen species (ROS) and lipid peroxidation, while it increased the Gamma-aminobutyric acid receptor subunit-ɑ1 (GABAAR-ɑ1) level as well as the activity of antioxidant enzymes superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx). Furthermore BME reduced the levels of HIF-1α and its downstream targets glucose transporter-1 (GLUT-1) and erythropoietin (EPO) in the DG, CA1, and CA3 regions of hippocampus. Additionally, results obtained from the open field, elevated zero maze and plus maze tests indicate that BME restores anxiety caused by hypoxia. Together, these findings suggested that BME mitigates the harmful effects of oxidative stress and altered hypoxia related signaling in hippocampus; and may provide a basis for its therapeutic use in the recovery from hypoxia-led anxiety.

Gadopiclenol Enables Reduced Gadolinium Dose While Maintaining Quality of Pulmonary Arterial Enhancement for Pulmonary MRA: An Opportunity for Improved Safety and Sustainability.

Pulmonary magnetic resonance angiography (MRA) is an imaging method with proven utility for the exclusion of pulmonary embolism and avoids the need for ionizing radiation and iodinated contrast agents. High-relaxivity gadolinium-based contrast agents (GBCAs), such as gadopiclenol, can be used to reduce the required gadolinium dose for pulmonary MRA. The aim of this study was to compare the contrast enhancement performance of gadopiclenol with an established gadobenate dimeglumine-enhanced pulmonary MRA protocol. In this retrospective single-center study, data from 152 patients who underwent pulmonary MRA at 1.5 T were analyzed. Imaging was performed with either 0.05 mmol/kg gadopiclenol (n = 75) or 0.1 mmol/kg gadobenate dimeglumine (n = 77), using dedicated multiphasic imaging protocols with precontrast, pulmonary arterial phase, immediate delayed phase, and a low flip-angle T1-weighted spoiled gradient echo acquisition. Subjective image quality evaluation was performed blinded by 2 radiologists on a 5-point Likert scale. For the estimation of interrater reliability, Cohen weighted κ was calculated. For semiquantitative assessment, signal intensities were measured in the pulmonary arteries, and relative signal enhancement was calculated. Data from groups were compared with Mann-Whitney U tests using Bonferroni corrections. Signal enhancement relative to precontrast in the first-pass pulmonary arterial phase was higher with 0.05 mmol/kg gadopiclenol compared with 0.1 mmol/kg gadobenate dimeglumine (20.0-fold ± 5.6-fold vs 17.8-fold ± 5.8-fold; P = 0.015). Readers observed no difference in subjective rating in terms of intravascular contrast, peripheral vessel depiction, and diagnostic confidence with substantial interrater reliability (Cohen κ = 0.73 [95% confidence interval: 0.57-0.89], 0.65 [0.55-0.75], and 0.74 [0.65-0.84], all P's < 0.001). No severe adverse events were recorded for any clinical MRA examination. The high-relaxivity contrast agent gadopiclenol can facilitate a reduction in gadolinium dose by 50% without compromising contrast enhancement for pulmonary MRA. This approach may enhance the safety and sustainability of pulmonary MRA in the long term.

Exploring the Mechanism of Shi-San-Wei-He-Zhong-Wan in the Treatment of Functional Dyspepsia Based on Network Pharmacology and Experimental Validation.

The incidence of Functional Dyspepsia (FD) is gradually increasing, yet there are currently no effective treatment methods available. This study explored the effective components, potential targets, and pathways of Shi-San-Wei-He-Zhong-Wan (SSWHZW) in the treatment of FD, aiming to provide new insights into its treatment. First, the Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and GeneCards databases were utilized to identify the major active components of SSWHZW and potential therapeutic targets of FD. Subsequently, functional enrichment analyses were performed to elucidate the mechanisms of SSWHZW on FD. Molecular docking simulations were then conducted to assess the binding affinity of key targets and major active components. Next, an FD animal model was established, and the therapeutic effects of SSWHZW were validated using Hematoxylin and Eosin (HE) staining and Enzyme-linked Immunosorbent Assay (ELISA). Finally, Western blot analysis was performed to validate the involvement of key signaling pathways. A total of 229 active ingredients and 283 putative targets were identified from SSWHZW, of which 173 overlapped with the targets of FD and were considered potential therapeutic targets. Key ingredients, such as quercetin, kaempferol, wogonin, and baicalein, were identified as pivotal components of SSWHZW, potentially acting on the 173 overlapping targets and influencing FD through related signaling pathways. Functional enrichment analysis revealed that the PI3K-Akt signaling pathway, VEGF signaling pathway, and NF-kappa B signaling pathway may be involved in the mechanism of SSWHZW in treating FD. Molecular docking predicted that all five ingredients could firmly bind with the top-ranked target TP53 in the Protein- protein Interaction (PPI) network. Further experiments demonstrated that SSWHZW protected the intestinal tissues of FD rats from inflammatory damage by inhibiting the PI3K/AKT signaling pathway. Based on network pharmacology, this study explored the multi-component, multitarget, and multi-pathway characteristics of SSWHZW in treating FD. The findings suggest that SSWHZW exerts its anti-FD effects by inhibiting the expression of the PI3K/AKT signaling pathway, providing new insights and methods for further research on the mechanism of SSWHZW in treating FD.

Hepatobiliary Adverse Events Associated With the KRAS p.G12C Inhibitor Sotorasib.

The p.G12C mutation in KRAS is commonly found in many cancers and was previously untreatable until drugs like sotorasib were developed. However, up to 15% of patients treated with sotorasib have experienced hepatobiliary adverse events. To investigate whether these side effects are more common among sotorasib users, a pharmacovigilance study is necessary. This study used the FDA adverse event reporting system (FAERS) database, a publicly available repository of reported drug adverse events, and AERSMine, an open-access pharmacovigilance tool, to investigate these adverse events. A total of 428 hepatobiliary adverse events were linked to sotorasib. Hepatic cytolysis had the highest reported relative risk at 26.541 and a safety signal of 4.726. Elevated liver and biliary enzymes such as AST, ALT, ALP, and GGT were commonly observed, but with lower reported relative risk and safety signal values, which supports previous real-world reports. These findings highlight the hepatobiliary risks associated with sotorasib and underscore the importance of closely monitoring liver function in patients who are using the medication. This is particularly crucial for patients with hepatobiliary cancers, as disease progression and adverse events could be misinterpreted.

Network Pharmacology, Molecular Docking, Molecular Dynamics to Explore the Mechanism of Danggui Shaoyao Powder for Hepatic Encephalopathy.

Patients with hepatic encephalopathy (HE) have many triggers and a high mortality rate. The protective effect of existing therapeutic drugs on the liver is weak. We found that Danggui Shaoyao Powder can improve the symptoms of HE and may have a better liver protection effect. And the mechanism of it is unclear. The research explores the mechanism of Danggui Shaoyao Powder for the treatment of HE through network pharmacology, molecular docking and molecular dynamics. Targets of Danggui Shaoyao Powder were screened from Traditional Chinese Medicine System Pharmacology Platform (TCMSP), SwissTargetPrediction, and Uniport. GeneCards was used to gain targets of HE. Further, core targets and ingredients were screened by protein-protein interaction network (PPI) and herbs-compounds-targets network. Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were completed to screen relative sites and signaling pathways. Molecular docking and dynamics were used to show the stability of ligand-receptor complexes. IL6, SRC and kaempferol, beta-sitosterol were screened as the top two core targets and ingredients. Dendrites, dendritic trees, and membrane sides were defined as the main sites of action. Core signaling pathways were screened such as: PI3K-Akt and MAPK. Molecular docking shows well-defined binding sites and the stability of the binding is demonstrated by molecular dynamics. Through this study, Danggui Shaoyao Powder may act on IL6, SRC, and other targets through ingredients such as kaempferol and beat-sitosterol and regulate signaling pathways such as PI3K-Akt, MAPK and NF-κB to the treatment of HE.