Background Asthma is a chronic inflammatory airway disease characterized by Th2- dominant immune responses, airway hyperresponsiveness, and structural remodeling. Although inhaled corticosteroids and biologics are effective for many patients, a substantial proportion remains poorly controlled and experiences treatment-related adverse effects. Probiotics have emerged as immunomodulatory agents in asthma, but existing studies predominantly focus on oral administration and gut-lung axis regulation. Whether direct respiratory administration of probiotics can modulate the pulmonary immune microenvironment and alleviate asthma remains largely unexplored. Methods An ovalbumin (OVA)-induced asthma model was established in C57BL/6 J mice. Clostridium butyricum, Lactobacillus casei, or Bifidobacterium infantis were administered intranasally during the challenge phase. Inflammation and remodeling were evaluated using bronchoalveolar lavage fluid (BALF) cell counts, serum IgE measurement, and histological staining (H&E, PAS, Masson). Lung immune and stromal cell heterogeneity and intercellular interactions were analyzed by single-cell RNA sequencing, while airway microbiota composition was characterized by 16S rDNA sequencing. Results Intranasal probiotic administration attenuated OVA-induced airway hyperresponsiveness, eosinophilic inflammation, and was associated with reduced mucus hypersecretion and collagen-associated histological changes. Probiotics suppressed Th2-biased immune responses, evidenced by reduced Th2 cell proportions, downregulation of Gata3, and decreased expression of Il4, Il5, and Il13. These effects were associated with inhibited dendritic cell activation and weakened DC-T cell interactions, particularly via the Cxcl16-Cxcr6 axis. Probiotic treatment was associated with an increased proportion of M2 macrophages, reduced pro-inflammatory signaling, reduced predicted macrophage-fibroblast communication through the Osm-Osmr pathway, and a lower proportion of fibrotic fibroblasts. Additionally, intranasal probiotic treatment was associated with changes in airway microbial composition, including a reduced relative abundance of Neisseria, which was positively correlated with Th2 cytokine expression. Conclusion Intranasal probiotic administration was associated with attenuation of airway inflammation and early remodeling-associated changes, accompanied by coordinated changes in Th2 immunity, dendritic cell activation, macrophage polarization, fibroblast subtypes, and the airway microbiota. These effects appeared to be strain-specific, with Lactobacillus casei, Bifidobacterium infantis, and Clostridium butyricum being predominantly associated with suppression of Th2 immunity, modulation of predicted DC-T cell communication, and macrophage polarization, respectively. These findings provide preclinical evidence supporting further investigation of intranasal probiotics as a potential strategy for asthma management.

Photoaging, driven by chronic UVR, disrupts skin structure and function. Traditional retinoids enhance extracellular matrix regeneration but cause irritation. Hydroxypinacolone 9-cis retinoate (9-cis HPR), a derivative of 9-cis retinoic acid, selectively activates retinoic acid receptor α and retinoid X receptor α, improving efficacy and tolerability. In a UVR-induced SKH-1 mouse photoaging model, 9-cis HPR reduced erythema, desquamation, and loss of elasticity while promoting collagen and elastin production. Single-cell RNA sequencing and spatial transcriptomics revealed restoration of fibroblast, basal cell, and melanocyte proportions; suppression of myofibroblast differentiation; and upregulation of extracellular matrix-related genes (eg, Col1a2, Col3a1, Elastin). In addition, 9-cis HPR inhibited melanogenesis by downregulating melanogenesis-related genes (Tyr, Dct, Tyrp1), melanosome biogenesis genes (Mlana, Pmel), and the melanocyte proliferation gene Kit, likely through ROS suppression. Cell-cell interaction analysis showed that 9-cis HPR promoted fibroblast-driven repair through neuropeptide Y-NPY1R, PTN-SDC2, and POSTN-ITGA/BV signaling, while inhibiting KITL-KIT-mediated melanocyte proliferation. In a single-blind, split-face clinical trial involving 31 Chinese women, 0.03% 9-cis HPR applied daily for 4 weeks demonstrated comparable or superior improvements in wrinkles, elasticity, hydration, dermal density, and radiance versus 0.3% retinol, without observed irritation. These findings support 9-cis HPR as a safe and effective retinoid that mitigates photoaging through extracellular matrix restoration, inflammation modulation, and pigmentation control.

The number of goblet cells is an important biomarker for the diagnosis and prognosis of asthma disease. However, visual assessment of a plethora of goblet cells is laborious and time-consuming, making it difficult to precisely identify and count the number of the cells in pathohistological slides. We developed an automated method for identifying goblet cells. It was a combination of coarse and refined recognition, including extraction of the cell region and separation of the adjacent cells. The goblet cell region was extracted with the superpixel method. Initial segmentation results were obtained by merging over-segmented superpixels based on their average intensities in the CIE L*a*b* color space. After that, in refining recognition process, the iCut algorithm was applied to segment touching cells and identify individual goblet cells. The proposed method was evaluated on the images of the lung tissue in a rat model of asthma. With a precision of 84.9 +/- 4.2 % and the F1-score of 82.8 +/- 4.0 %, the proposed method surpassed the other four conventional cell recognition algorithms on the both metrics with relatively higher computational cost. This work demonstrated precise identification of goblet cells based on histopathological slides, which is crucial for evaluating the extent of asthma, enabling investigation into the disease's origin and the efficacy of related medications. It can also be flexibly extended to cell segmentation in other biomedical images.

Psoriasis is an incurable chronic inflammatory disease that requires new interventions. Here, we found that fibroblasts exacerbate psoriasis progression by promoting macrophage recruitment via CCL2 secretion by single-cell multi-omics analysis. The natural small molecule celastrol was screened to interfere with the secretion of CCL2 by fibroblasts and improve the psoriasis-like symptoms in both murine and cynomolgus monkey models. Mechanistically, celastrol directly bound to the low-density lipoprotein receptor-related protein 1 (LRP1) β-chain and abolished its binding to the transcription factor c-Jun in the nucleus, which in turn inhibited CCL2 production by skin fibroblasts, blocked fibroblast–macrophage crosstalk, and ameliorated psoriasis progression. Notably, fibroblast-specific LRP1 knockout mice exhibited a significant reduction in psoriasis like inflammation. Taken together, from clinical samples and combined with various mouse models, we revealed the pathogenesis of psoriasis from the perspective of fibroblast-macrophage crosstalk, and provided a foundation for LRP1 as a novel potential target for psoriasis treatment.

BACKGROUND:Single-cell RNA sequencing (scRNA-seq) has become a significant tool for addressing complex issuess in the field of biology. In the context of scRNA-seq analysis, it is imperative to accurately determine the type of each cell. However, conventional supervised or semi-supervised methodologies are contingent on expert labels and incur substantial labeling costs, In contrast self-supervised pre-training strategies leverage unlabeled data during the pre-training phase and utilise a limited amount of labeled data in the fine-tuning phase, thereby greatly reducing labor costs. Furthermore, the fine-tuning does not need to learn the feature representations from scratch, enhancing the efficiency and transferability of the model. METHODS:The proposed methodology is outlined below. The deep learning framework, TransAnno-Net, is based on transfer learning and a Transformer architecture. It has been designed for efficient and accurate cell type annotations in large-scale scRNA-seq datasets of mouse lung organs. Specifically, TransAnno-Net is pre-trained on the scRNA-seq lung data of approximately 100,000 cells to acquire gene-gene similarities via self-supervised learning. It is then migrated to a relatively small number of datasets to fine-tune specific cell type annotation tasks. To address the issue of imbalance in cell types commonly observed in scRNA-seq data, we applied a random oversampling technique is applied to the fine-tuned dataset. This is done to mitigate the impact of distributional imbalance on the annotation outcomes. RESULTS:The experimental findings demonstrate that TransAnno-Net exhibits superior performance with an AUC of 0.979, 0.901, and 0.982, respectively, on three mouse lung datasets, outperforming eight state-of-the-art (SOTA) methods. In addition, TransAnno-Net demonstrates robust performance on cross-organ, cross-platform datasets, and is competitive with the fully supervised learning-based method. CONCLUSION:The TransAnno-Net method is a highly effective cross-platform and cross-data set single-cell type annotation method for mouse lung tissues and supports cross-organ cell type annotation. This approach is expected to enhance the efficiency of research on the biological mechanisms of complex biological systems and diseases.

Rationale: The hyperactivity of lateral habenula (LHb) has been implicated in the pathophysiology of depression, but the regulatory mechanisms of inhibitory synapses in this context remains unclear. MDGA1 and neuroligin2 (Nlgn2), both regulators of inhibitory synapses, selectively interact in the LHb. We aimed to investigate if their interaction contributes to chronic restrained stress (CRS)-induced depression by modulating inhibitory synapses. Methods: Transgenic mouse models were established to conditional knockout/recover of MDGA1 expression or knockin Nlgn2 variant incapable of binding MDGA1 in the LHb, using viral Cre-recombinase expression. Synaptic function and density were assessed through electrophysiology and immunostaining, respectively. An acute restrained stress (ARS) model and chemogenetic activation of the lateral hypothalamus (LH) were used to stimulate the LHb. Behavioral tests related to depression were conducted following CRS. Results: MDGA1 and Nlgn2 selectively interacted in the LHb, which was elevated following CRS. Germline knockout of MDGA1 increased inhibitory transmission and GABAergic synapse density in the LHb, effects that were reversed by adult re-expression of MDGA1. Introduction of the Nlgn2 variant incapable of binding MDGA1 similarly enhanced inhibitory transmission and increased GABAergic synapse density in the LHb. Both germline MDGA1 deficiency and introduction of the Nlgn2 variant mitigated ARS- and LH activation-induced LHb neuron hyperactivation. MDGA1 deficiency in the LHb during adulthood increased inhibitory synaptic strength and conferred significant resistance to CRS-induced depressive behaviors, similar to the effects of introducing the Nlgn2 variant in the LHb. Conclusions: Our findings suggests that MDGA1-mediated suppression of Nlgn2 facilitates depression onset through limiting GABAergic synapse formation within the LHb. Targeting MDGA1/Nlgn2 complexes residing at GABAergic synapses within the lateral habenula may be viable for alleviating core behavioral symptoms of major depression.

BACKGROUND:Eosinophilic chronic sinusitis (ECRS) is a refractory condition resistant to therapies and prone to relapse. Hexokinase 2 (HK2), a key glycolysis enzyme, regulates inflammation but its role in ECRS is unclear. METHODS:An ECRS mouse model was established using intranasal administration of papain. Single-cell RNA sequencing (scRNA-seq) was employed to analyze the expression patterns of Hk2 in ECRS. The HK2 inhibitor lonidamine (LND) was administered orally to assess symptoms, inflammatory cells and cytokines in the nasal lavage fluid (NALF), serum total IgE, and pathological characteristics of nasal mucosal inflammation. Mechanistic insights were investigated using scRNA-seq and an in vitro human nasal epithelial cells (HNEpC) model stimulated with IL-4, IL-13, and TNF-α. RESULTS:Elevated Hk2 expression was found in the nasal mucosa in the ECRS model. LND alleviated ECRS symptoms, reducing sneezing, cytokine release, inflammatory cell infiltration, and goblet cell hyperplasia. Epithelial cell damage was identified as a key driver of inflammation and remodeling. LND suppressed inflammation by inhibiting differentiation of inflammatory epithelial cells and neutrophils via Cxcl1-Cxcr2. Additionally, LND reduced epithelial cell-induced nasal mucosal remodeling by inhibiting the Ptn-Ncl signaling pathway between epithelial cells and neurons. In the in vitro experiment, LND significantly and dose-dependently reduced both CXCL1 and PTN expression, confirming its direct anti-inflammatory and anti-remodeling effects on nasal epithelial cells. CONCLUSIONS:LND significantly suppressed nasal inflammation and tissue remodeling in the ECRS model. These findings suggest that HK2 inhibition holds promise as a safe and effective therapeutic approach for the management of ECRS.

Pulmonary inflammation is closely associated with macrophage polarization and lipid metabolic reprogramming. Miconazole (MCZ), traditionally used as an antifungal agent, exhibits emerging anti-inflammatory potential, yet its underlying mechanisms remain unclear. A mouse model of lipopolysaccharide (LPS)-induced lung inflammation was employed to evaluate MCZ’s anti-inflammatory efficacy. In vivo inflammatory cell infiltration, cytokine expression (IL-6, IL-1β, TNF-α), and lung histopathology were assessed. Single-cell RNA sequencing (scRNA-seq) characterized alveolar macrophage subpopulations and associated lipid metabolism pathways. In vitro experiments with bone marrow-derived macrophages (BMDM) validated the changes of macrophage polarization. MCZ treatment significantly alleviated lung inflammation by decreasing inflammatory cell infiltration and suppressing pro-inflammatory cytokines. ScRNA-seq analysis revealed subcluster of Itgam (Cd11b) negative, Mrc1, Marco, and Lgals3 high AMs, MCZ decreased the proportions of pro-inflammatory neutrophils and macrophages, and promoted the phenotypic shift of alveolar macrophages from a pro-inflammatory subtype (AM1) to an anti-inflammatory subtype (AM2). Further cell-cell communication analysis showed that MCZ suppressed interactions between AM1 alveolar macrophages and neutrophils via TNF-TNFR, CCL3/5-CCR1, and CXCL1-CXCR2 signaling pathways. Mechanistically, MCZ inhibited lipid synthesis in AM1 alveolar macrophages while enhancing lipid catabolism in AM2 alveolar macrophages. In vitro studies using BMDM further confirmed that MCZ inhibited LPS-induced macrophage M1 polarization and lipid droplet accumulation marked by perilipin 3 (PLIN3), while promoting IL-4/IL-13-induced M2 polarization. MCZ exerts therapeutic effects against pulmonary inflammation primarily by modulating macrophage polarization through lipid metabolic reprogramming, highlighting its promise as a novel therapeutic approach for inflammatory lung diseases.

Rapamycin has been shown to be effective in the treatment of a variety of neurological disorders, including epilepsy. Intranasal drug administration is a novel mode of drug delivery that bypasses the blood-brain barrier and numerous biological effects thereby entering the central nervous system directly. Thus, the objective of this study was to investigate the brain entrance efficacy of rapamycin following intranasal administration of rapamycin.First, we found that acute high-dose administration with a total dose of 0.326 mg of rapamycin in a novel dosage form produced few side effects on body weight, various organs, and nasal mucosa in rats. Then, we examined the distribution of drug concentrations in the brain, nasal mucosa, and blood using the above dosage form administered intranasally to rats at 0.04 mg/kg. We found that intranasal administration was significantly more efficacious than oral administration for rapamycin brain delivery. We also discovered gender differences in drug absorption following intranasal administration of rapamycin, wherein rapamycin exhibited faster systemic absorption in female rats compared to males. Our study demonstrated that intranasal administration of rapamycin is highly effective and low toxic, which may provide a new delivery option for rapamycin therapy in brain diseases.

Acute lung injury (ALI) is an acute inflammatory lung disease associated with both innate and adaptive immune responses. Hexokinase 2 (HK2) is specifically highly expressed in numerous types of inflammation-related diseases and models. In the present study in vitro and in vivo effects of targeted degradation of HK2 on ALI were explored. The degradation of HK2 by the targeting peptide TAT (transactivator of transcription protein of HIV-1)-ataxin 1 (ATXN1)-chaperone-mediated autophagy-targeting motif (CTM) was demonstrated by ELISA and western blotting in vitro and in vivo. The inhibitory effects of TAT-ATXN1-CTM on lipopolysaccharide (LPS)-induced inflammatory responses were examined using ELISAs. The therapeutic effects of TAT-ATXN1-CTM on LPS-induced ALI were examined via histological examination and ELISAs in mice. 10 mu M TAT-ATXN1-CTM administration decreased HK2 protein expression and the secretion of proinflammatory cytokines (TNF-alpha and IL-1 beta) without altering HK2 mRNA expression in LPS-treated both in vitro and in vivo, while pathological lung tissue damage and the accumulation of leukocytes, neutrophils, macrophages and lymphocytes in ALI were also significantly suppressed by 10 mu M TAT-ATXN1-CTM treatment. TAT-ATXN1-CTM exhibited anti-inflammatory activity in vitro and decreased the severity of ALI in vivo. HK2 degradation may represent a novel therapeutic approach for ALI.

Synapse organizers are essential for the development, transmission, and plasticity of synapses. Acting as rare synapse suppressors, the MAM domain containing glycosylphosphatidylinositol anchor (MDGA) proteins contributes to synapse organization by inhibiting the formation of the synaptogenic neuroligin-neurexin complex. A previous analysis of MDGA2 mice lacking a single copy of Mdga2 revealed upregulated glutamatergic synapses and behaviors consistent with autism. However, MDGA2 is expressed in diverse cell types and is localized to both excitatory and inhibitory synapses. Differentiating the network versus cell-specific effects of MDGA2 loss-of-function requires a cell-type and brain region-selective strategy. To address this, we generated mice harboring a conditional knockout of Mdga2 restricted to CA1 pyramidal neurons. Here we report that MDGA2 suppresses the density and function of excitatory synapses selectively on pyramidal neurons in the mature hippocampus. Conditional deletion of Mdga2 in CA1 pyramidal neurons of adult mice upregulated miniature and spontaneous excitatory postsynaptic potentials, vesicular glutamate transporter 1 intensity, and neuronal excitability. These effects were limited to glutamatergic synapses as no changes were detected in miniature and spontaneous inhibitory postsynaptic potential properties or vesicular GABA transporter intensity. Functionally, evoked basal synaptic transmission and AMPAR receptor currents were enhanced at glutamatergic inputs. At a behavioral level, memory appeared to be compromised in Mdga2 cKO mice as both novel object recognition and contextual fear conditioning performance were impaired, consistent with deficits in long-term potentiation in the CA3-CA1 pathway. Social affiliation, a behavioral analog of social deficits in autism, was similarly compromised. These results demonstrate that MDGA2 confines the properties of excitatory synapses to CA1 neurons in mature hippocampal circuits, thereby optimizing this network for plasticity, cognition, and social behaviors.

BackgroundAs the largest organ of the body, the skin is constantly subjected to ultraviolet radiation (UVR), leading to inflammations and changes that mirror those seen in chronological aging. Although various small molecule drugs have been explored for treating skin photoaging, they typically suffer from low stability and a high incidence of adverse reactions. Consequently, the continued investigation of photoaging treatments, particularly those utilizing herbal products, remains a critical clinical endeavor. One such herbal product, Lapagyl, is derived from the bark of the lapacho tree and possesses antioxidant efficacies that could be beneficial in combating skin photoaging.PurposeThis research aimed to evaluate the efficacy of the herbal product Lapagyl in combating UVR-induced skin photoaging. Additionally, it sought to unravel the mechanisms by which Lapagyl promotes the regeneration of the skin extracellular matrix.MethodsTo investigate whether Lapagyl can alleviate skin aging and damage, a UVR radiation model was established using SKH-1 hairless mice. The dorsal skins of these mice were evaluated for wrinkle formation, texture, moisture, transepidermal water loss (TEWL), and elasticity. Pathological assessments were conducted to determine Lapagyl's efficacy. Additionally, single-cell sequencing and spectrum analysis were employed to elucidate the working mechanisms and primary components of Lapagyl in addressing UVR-induced skin aging and injury.ResultsLapagyl markedly reduced UVR-induced wrinkles, moisture loss, and elasticity decrease in SKH-1 mice. Single-cell sequencing demonstrated that Lapagyl corrected the imbalance in cell proportions caused by UVR, decreased UVR-induced ROS expression, and protected basal and spinous cells from skin damage. Additionally, Lapagyl effectively prevented the entry of inflammatory cells into the skin by reducing CCL8 expression and curtailed the UVR-induced formation of Foxp3+ regulatory T cells (Tregs) in the skin. Both pathological assessments and ex vivo skin model results demonstrated that Lapagyl effectively reduced UVR-induced damage to collagen and elastin. Spectrum analysis identified Salidroside as the primary compound remaining in the skin following Lapagyl treatment. Taken together, our study elucidated the skin protection mechanism of the herbal product Lapagyl against UVR damage at the cellular level, revealing its immunomodulatory effects, with salidroside identified as the primary active compound for skin.ConclusionOur study provided a thorough evaluation of Lapagyl's protective effects on skin against UVR damage, delving into the mechanisms at the cellular level. We discovered that Lapagyl mitigates skin inflammation and immunosuppression by regulating Foxp3+ Tregs and the CCL pathway. These insights indicate that Lapagyl has potential as a novel therapeutic option for addressing skin photoaging.

Background:Lung inflammation occurs in many lung diseases, but has limited effective therapeutics. Ginseng and its derivatives have anti-inflammatory effects, but their unstable physicochemical and metabolic properties hinder their application in the treatment. Panaxadiol (PD) is a stable saponin among ginsenosides. Inhalation administration may solve these issues, and the specific mechanism of action needs to be studied.Methods:A mouse model of lung inflammation induced by lipopolysaccharide (LPS), an in vitro macrophage inflammation model, and a coculture model of epithelial cells and macrophages were used to study the effects and mechanisms of inhalation delivery of PD. Pathology and molecular assessments were used to evaluate efficacy. Transcriptome sequencing was used to screen the mechanism and target. Finally, the efficacy and mechanism were verified in a human BALF cell model.Results:Inhaled PD reduced LPS-induced lung inflammation in mice in a dose-dependent manner, including inflammatory cell infiltration, lung tissue pathology, and inflammatory factor expression. Meanwhile, the dose of inhalation was much lower than that of intragastric administration under the same therapeutic effect, which may be related to its higher bioavailability and superior pharmacokinetic parameters. Using transcriptome analysis and verification by a coculture model of macrophage and epithelial cells, we found that PD may act by inhibiting TNFA/TNFAR and IL7/IL7R signaling to reduce macrophage inflammatory factor-induced epithelial apoptosis and promote proliferation.Conclusion:PD inhalation alleviates lung inflammation and pathology by inhibiting TNFA/TNFAR and IL7/IL7R signaling between macrophages and epithelial cells. PD may be a novel drug for the clinical treatment of lung inflammation.

Background Suboptimal adherence to asthma medication is one of the major causes of asthma exacerbation. Currently, there is no report on the objective and scientific evaluation of medication adherence among asthmatic children. We aimed to identify and quantify adherence and techniques of using dry powder inhalers (DPIs) among pediatric asthmatic patients with a newly developed electronic device that can monitor the use pattern of DPIs during inhalation. Methods A prospective, single-center, observational cohort study was conducted. On discharge from the hospital, pediatric asthmatic patients were given Diskus DPIs with attached electronic devices. The frequency, flow velocity, volume, duration and angle were analyzed to determine the adherence and technical proficiency of DPI use. Results Pediatric asthmatic patients (n = 128) with a mean age of 7.8 years, a mean forced expiratory volume in 1 second (FEV1) of 1.5 L, and a mean FEV1% of 93.5% were recruited. Total 4096 results were recorded. The most common error types were short duration (98.8%), low volume (94.5%), wrong angle (33.3%), missed use (27.9%), low peak inspiratory flow rate (PIFR) (24.4%), exhalation (15.8%) and multiple use (0.8%). The mean actual adherence was 0.1% (standard deviation (SD), 0.9%). The errors in Diskus DPI use were correlated with asthmatic children's age, basic lung function, and desensitization treatment status. Conclusion Our study is the first to objectively characterize the primary types and proportions of technique errors among asthmatic children using modified Diskus DPIs and revealed that asthmatic children’s real-world actual adherence to DPIs was unsatisfactory.

BACKGROUND:Idiopathic pulmonary fibrosis (IPF) stands as a significant contributor to global mortality rates. Presently, there exists a dearth of effective anti-fibrotic treatments for this condition. While itraconazole (ITR) has exhibited potential in mitigating pulmonary fibrosis, its oral administration is hampered by unfavorable pharmacokinetics, which elevate the risk of adverse reactions, thus limiting its clinical utility. METHODS:An inhalable formulation of ITR were engineered which aimed at enhancing its pulmonary dispersion. First, pharmacokinetics were conducted to investigate the blood concentration and tissue residue of ITR after inhalation administration. In addition, bleomycin induced mouse pulmonary fibrosis model was used to compare the therapeutic effects of ITR administered by inhalation and intragastric administration. Finally, single-cell RNA sequencing (scRNAseq) was used to explore the mechanism of ITR inhalation administration. RESULTS:We found that a large amount of drugs accumulated in the lung tissue for a long time after inhalation administration, thus maximizing the therapeutic effect of drugs. Inhalation of ITR daily at for 21 days significantly attenuated bleomycin-induced lung fibrosis and inflammation in murine models. Additionally, our findings revealed that ITR inhalation diminished the proportion of diseased fibroblasts while promoting reparative fibroblast populations in the murine model. Furthermore, it effectively reversed the proportion of activated phagocytic macrophages. Mechanistically, ITR inhalation exerted its effects by regulating SPP1 and C3 signaling pathway pivotal in the interaction between phagocytic macrophages and diseased fibroblasts. CONCLUSIONS:These insights into the molecular mechanisms underlying ITR's therapeutic effects on IPF underscore the favorable pharmacokinetic profile conferred by inhalation, thus presenting a promising formulation poised for clinical translation.

Awareness of estrogen's effects on health is broadening rapidly. The effects of long-term high levels of estrogen on the body involve multiple organs. Here, we used both single-cell chromatin accessibility and RNA sequencing data to analyze the potential effect of estrogen on major organs. The integrated cell map enabled in-depth dissection and comparison of molecular dynamics, cell-type compositions, and cellular heterogeneity across multiple tissues and organs under estrogen stimulation. We also inferred pseudotime cell trajectories and cell-cell communications to uncover key molecular signatures underlying their cellular processes in major organs in response to estrogen. For example, estrogen could induce the differentiation of IFIT3+ neutrophils into S100A9+ neutrophils involved in the function of endosome-to-lysosome transport and the multivesicular body sorting pathway in liver tissues. Furthermore, through integration with human genome-wide association study data, we further identified a subset of risk genes during disease development that were induced by estrogen, such as AKT1 (related to endometrial cancer), CCND1 (related to breast cancer), HSPH1 (related to colorectal cancer), and COVID-19 and asthma-related risk genes. Our work uncovers the impact of estrogen on the major organs, constitutes a useful resource, and reveals the contribution and mechanism of estrogen to related diseases.

Intracerebral hemorrhage (ICH) can induce intensive oxidative stress, neuroinflammation, and brain cell apoptosis. However, conventional methods for ICH treatment have many disadvantages. There is an urgent need for alternative, effective therapies with minimal side effects. Pharmacodynamics experiment, molecular docking, network pharmacology, and metabolomics were adopted to investigate the treatment and its mechanism of Jingfang Granules (JFG) in ICH. In this study, we investigated the therapeutic effects of JFG on ICH using behavioral, brain water content and Magnetic resonance imaging experiments. However, the key active component and targets of JFG remain unknown. Here we verified that JFG was beneficial to improve brain injury after ICH. A network pharmacology analysis revealed that the anti-inflammatory effect of JFG is predominantly mediated by its activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway through Luteolin, (+)-Anomalin and Phaseol and their targeting of AKT1, tumor necrosis factor alpha (TNF-alpha), and interleukin-1 beta (IL-1 beta). Molecular docking analyses revealed an average affinity of -8.633 kcal/mol, indicating a binding strength of less than -5 kcal/mol. Metabolomic analysis showed that JFG exerted its therapeutic effect on ICH by regulating metabolic pathways, such as the metabolism of taurine and hypotaurine, biosynthesis of valine, leucine, and isoleucine. In conclusion, we demonstrated that JFG attenuated neuroinflammation and BBB injury subsequent to ICH by activating the PI3K/Akt signaling pathway.

目的 研究星形胶质细胞大麻素1(CB1)受体参与调控海马中间神经元长时程增强的机制.方法 ①采用穿孔全细胞膜片钳的方法记录海马中间神经元长时程增强.②使用药理学手段阻断星形胶质细胞Ca2+信号,新陈代谢或CB1受体观察对海马中间神经元长时程增强的影响.③使用Ca2+成像技术和膜片钳技术分别观察星形胶质细胞Ca2+信号机制和长时程增强的分子机制.结果 阻断星形胶质细胞CB1受体、螯合Ca2+信号和抑制星形胶质细胞新陈代谢均导致海马中间神经元长时程增强受损;Ca2+成像结果表明在诱导长时程增强时星形胶质细胞Ca2+信号升高是通过激活CB1受体引起的;电生理突触机制分析表明星形胶质细胞通过释放D-丝氨酸结合突触后NMDA受体调控海马中间神经元长时程增强.结论 星形胶质细胞CB1受体激活引起Ca2+信号增强,引起D-丝氨酸释放,D-丝氨酸结合NMDA受体调控海马中间神经元长时程增强.

Background: Aggressive behaviors are one of the most important negative behaviors that seriously endangers human health. Also, the central para-inflammation of microglia triggered by stress can affect neurological function, plasticity, and behavior. NLRP3 integrates stress-related signals and is a key driver of this neural para-inflammation. However, it is unclear whether the NLRP3 inflammasome is implicated in the development of aggressive behaviors. Methods: First, aggressive behavior model mice were established using the resident intruder paradigm. Then, aggressive behaviors were determined with open-field tests (OFT), elevated plus-maze (EPM), and aggressive behavior tests (AT). Moreover, the expression of P2X7R and NLRP3 inflammasome complexes were assessed by immunofluorescence and Western blot. The levels of NLRP3 and inflammatory cytokines were evaluated using enzyme-linked immunosorbent assay (ELISA) kits. Finally, nerve plasticity damage was observed by immunofluorescence, transmission electron microscope, and BrdU staining. Results: Overall, the resident intruder paradigm induced aggressive behaviors, activated the hippocampal P2X7R and NLRP3 inflammasome, and promoted the release of proinflammatory cytokines IL-1β in mice. Moreover, NLRP3 knockdown, administration of P2X7R antagonist (A804598), and IL-1β blocker (IL-1Ra) prevented NLRP3 inflammasome-driven inflammatory responses and ameliorated resident intruder paradigm-induced aggressive behaviors. Also, the resident intruder paradigm promoted the activation of mouse microglia, damaging synapses in the hippocampus, and suppressing hippocampal regeneration in mice. Besides, NLRP3 knockdown, administration of A804598, and IL-1Ra inhibited the activation of microglia, improved synaptic damage, and restored hippocampal regeneration. Conclusion: The NLRP3 inflammasome-driven inflammatory response contributed to resident intruder paradigm-induced aggressive behavior, which might be related to neuroplasticity. Therefore, the NLRP3 inflammasome can be a potential target to treat aggressive behavior-related mental illnesses.