Background The contamination of public baths with Legionella pneumophila contamination has become a growing public health concern in recent years. However, research on its association with bacterial community characteristics in water samples remains limited. The integration of 16S rRNA sequencing and random forest modeling provides a new approach to elucidate the bacterial community characteristics of public bath water and their association with Legionella pneumophila contamination. Objective To investigate the bacterial community structure and diversity of public bath water in Shanghai, explore the association between Legionella pneumophila contamination and bacterial community characteristics, and identify key bacterial genera associated with contamination, thereby providing a scientific basis for formulating hygiene management regulations for public bath water. Methods From February to March 2023, water samples were collected from ten public baths in Shanghai which were selected based on business scale, regional distribution, and functional differences. Water quality parameters were evaluated, and the samples were categorized into Legionella-positive and Legionella-negative groups based on the detection results of Legionella pneumophila. The bacterial community structure, α-diversity, and β-diversity were analyzed using 16S rRNA sequencing. Redundancy analysis (RDA) was employed to examine the relationship between physicochemical factors and bacterial community diversity. A random forest model was employed to identify key bacterial genera distinguishing the two groups, with the importance of genera being evaluated based on the mean decrease accuracy (MDA). Results The oxygen consumption in the Legionella-positive group was significantly lower than that in the Legionella-negative group (mean values: 1.85 mg·L⁻¹ vs. 6.81 mg·L⁻¹, P< 0.05), while no significant differences were observed in other physicochemical indicators. The sequencing results revealed a total of 27 bacterial phyla and 454 bacterial genera, with Proteobacteria (63.00%) being the dominant phylum. The dominant genera included Pelomonas (8.50%), Acidovorax (8.13%), Mycobacterium (7.93%), and Acinetobacter (6.59%). The α-diversity analysis indicated that bacterial community richness (Chao1 and ACE indices) was significantly higher in the Legionella-positive group than in the Legionella-negative group (P<0.01). The β-diversity analysis showed no significant difference in the bacterial community structure between the two groups (P>0.05). The RDA analysis demonstrated that the bacterial community diversity was positively correlated with pH and negatively correlated with oxygen consumption and free residual chlorine. The RDA1 and RDA2 explained 23.92% and 21.30% of the bacterial community diversity, respectively. The random forest model identified 20 key genera significantly influencing the microbial community distribution between the two groups, including unclassified_Bradyrhizobiaceae (MDA=2.42), Meiothermus (MDA=2.37), and Flavihumibacter (MDA=2.26). Conclusion The diversity of bacterial communities in public bath water is influenced by pH, oxygen consumption, and free residual chlorine. Samples contaminated with Legionella pneumophila exhibit greater microbial richness and contain characteristic key bacterial genera that contribute to community differences. Machine learning random forest technology helps identify these distinctive key bacterial genera. The findings provide a basis for carrying out risk early warning strategies in such settings.

Maternal obesity has been shown to exert adverse transgenerational effects on offspring ovarian function,characterized by reduced ovarian follicle reserve,impaired granulosa cell function,and disrupted reproductive cycles.The underlying mechanisms involve cellular stress-induced damage characterized by mitochondrial dysfunction,oxidative stress,and apoptosis,as well as dysregulation in key regulatory pathways such as ovarian hormone secretion,gene expression,and epigenetic modifications.This review synthesizes existing evidence on how maternal obesity-induced acute stress and chronic genetic/epigenetic alterations jointly contribute to offspring ovarian impairment.The findings elu-cidate the complex relationship between maternal metabolic status and offspring reproductive health,providing a robust theoretical foundation for refining prenatal management and developing targeted intervention strategies.

Traumatic brain injury (TBI) involves both primary mechanical damage and refractory secondary injuries, resulting in high disability rate and poor prognosis. Current therapeutic strategies for TBI include surgical intervention, neuroprotective agents, moderate hypothermia therapy and spinal cord stimulation. However, most of these therapeutic approaches primarily address wound surface management rather than targeting specific pathogenic mechanisms underlying post-injury inflammation and neurodegenerative diseases, resulting in suboptimal efficacy. Consequently, novel therapeutic strategies targeting TBI pathological mechanisms are urgently needed. The endogenous cannabinoid system (ECS) exerts multifaceted neuroprotective effects in TBI by modulating neuroinflammation, inhibiting glutamate excitotoxicity and activating pathways such as phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt). Investigating the characteristics of ECS components and their related signaling pathways may yield new approaches in the development of neuroprotective drugs for TBI. Nevertheless, few ESC-targeting drugs for TBI treatment have advanced beyond preclinical or clinical trial phases. Breakthroughs in this field depend on a deeper understanding of ECS and its mechanisms in TBI. To this end, the authors reviewed researches on the composition and functions of ECS, as well as the mechanisms underlying its neuroprotective effects following TBI, aiming to provide references for the development of ECS-targeting therapies.

Aim To explore the effects of exercise during pregnancy on renal structure,function and angiotensin Ⅱ(Ang Ⅱ)/transforming growth factor-β1(TGF-β1)/connective tissue growth factor(CTGF)signaling pathway in 3-month-old offspring of spontaneously hypertensive rats(SHR),the aim of this study was to provide experimental basis for early intervention of hypertension and protection of key target organs.Methods After mating SHR and WKY rats,pregnant rats were randomly divided into sedentary group(p-WKY-SED,p-SHR-SED)and exercise group(p-WKY-EX,p-SHR-EX).Blood pressure,serum urea nitrogen and creatinine were measured by caudal artery non-invasive blood pressure system and colorimetry in 3-month-old offspring rats.HE staining,Masson staining,ELISA and Western blot were used to detect the renal structure,collagen volume fraction,Ang Ⅱ concentration,renin-angiotension-aldosterone sys-tem(RAAS)and protein expression related to fibrogenic signal pathway in 3-month-old rats.Results(1)The sys-tolic blood pressure(SBP),diastolic blood pressure(DBP)and mean arterial pressure(MAP)of offspring rats in p-SHR-SED group were significantly higher than those in p-WKY-SED group.The SBP,DBP and MAP of SHR male off-spring rats were significantly decreased by exercise during pregnancy(P＜0.05),but had no effect on the female offspring rats(P＞0.05).(2)There was no significant difference in serum urea nitrogen and creatinine among the groups(P＞0.05).(3)The glomerular volume and the collagen volume fraction in p-SHR-SED group were significantly higher than those in p-WKY-SED group(P＜0.05),and the glomerular volume and the collagen volume fraction in p-SHR-EX group were significantly lower than those in p-SHR-SED group(P＜0.05).(4)Renal Ang Ⅱ level of offspring rats in p-SHR-SED group was significantly higher than that in p-WKY-SED group,and renal Ang Ⅱ level of offspring rats in p-SHR-EX group was significantly lower than that in p-SHR-SED group(P＜0.05).(5)The expression levels of angiotensin Ⅱ type 1 receptor(AT1R),TGF-β1 and CTGF protein in p-SHR-SED group were significantly higher than those in p-WKY-SED group(P＜0.05),while the expression levels of angiotensin converting enzyme 2(ACE2),angiotensin Ⅱ type 2 receptor(AT2R)and MasR protein in p-SHR-SED group were significantly lower than those in p-WKY-SED group(P＜0.05).Conclusion(1)Exercise during pregnancy can significantly decrease the blood pressure of 3-month-old male offspring rats of hypertensive rats,but has no significant effect on that of 3-month-old female offspring.(2)Exercise during preg-nancy may reduce renal fibrosis in 3-month-old female/male offspring of hypertensive rats by regulating RAAS balance and inhibiting Ang Ⅱ/TGF-β1/CTGF signaling pathway.

Given that ovarian stimulation is vital for assisted reproductive technology (ART) and results in elevated serum estrogen levels, exploring the impact of elevated estrogen exposure on oocytes and embryos is necessary. We investigated the effects of various ovarian stimulation treatments on oocyte and embryo morphology and gene expression using a mouse model and estrogen-treated mouse embryonic stem cells (mESCs). Female C57BL/6J mice were subjected to two types of conventional ovarian stimulation and ovarian hyperstimulation; mice treated with only normal saline served as controls. Hyperstimulation resulted in high serum estrogen levels, enlarged ovaries, an increased number of aberrant oocytes, and decreased embryo formation. The messenger RNA (mRNA)‍-sequencing of oocytes revealed the dysregulated expression of lysine-specific demethylase 6b (Kdm6b), which may be a key factor indicating hyperstimulation-induced aberrant oocytes and embryos. In vitro, Kdm6b expression was downregulated in mESCs treated with high-dose estrogen; treatment with an estrogen receptor antagonist could reverse this downregulated expression level. Furthermore, treatment with high-dose estrogen resulted in the upregulated expression of histone H3 lysine 27 trimethylation (H3K27me3) and phosphorylated H2A histone family member X (γ-H2AX). Notably, knockdown of Kdm6b and high estrogen levels hindered the formation of embryoid bodies, with a concomitant increase in the expression of H3K27me3 and γ-H2AX. Collectively, our findings revealed that hyperstimulation-induced high-dose estrogen could impair the demethylation of H3K27me3 by reducing Kdm6b expression. Accordingly, Kdm6b could be a promising marker for clinically predicting ART outcomes in patients with ovarian hyperstimulation syndrome.
Female ; Mice ; Demethylation/drug effects* ; Embryonic Stem Cells ; Estrogens/administration & dosage* ; Gene Expression/drug effects* ; Histones/metabolism* ; Jumonji Domain-Containing Histone Demethylases/metabolism* ; Mice, Inbred C57BL ; Oocytes ; Ovary/drug effects* ; Reproductive Techniques, Assisted ; Animals

Objective To compare the efficacy and safety of computed tomography (CT)-guided percutaneous versus electromagnetic navigation bronchoscopy (ENB)-guided microwave ablation (MWA) for the treatment of pulmonary nodules. Methods A retrospective analysis was conducted on the data of high-risk pulmonary nodule patients who underwent MWA at the Nanjing Drum Tower Hospital between 2022 and 2023. The pathological diagnosis rate, complications, and progression-free survival (PFS) rate were compared between the CT group and the ENB group. Results There were 61 patients in the CT group, including 30 males and 31 females, with an average age of (67.22±9.13) years. There were 53 patients in the ENB group, including 29 males and 24 females, with an average age of (65.29±13.76) years. The pathological diagnosis rate in the CT group was slightly higher than that in the ENB group (88.52% vs. 71.69%, P=0.03). However, the ENB group exhibited a lower incidence of perioperative complications, including pneumothorax (16.39% vs. 3.77%, P=0.03), hemoptysis (19.67% vs. 5.66%, P=0.05), and pain (22.95% vs. 7.55%, P=0.03). There was no statistically significant difference in PFS rate between the two groups [HR=1.17, 95%CI (0.23, 5.81), P=0.85]. Conclusion Both CT-guided and ENB-guided MWA are effective treatment modalities for high-risk pulmonary nodules.

Environmental toxicants have been linked to aging and age-related diseases. The emerging evidence has shown that the enhancement of detoxification gene expression is a common transcriptome marker of long-lived mice, Drosophila melanogaster, and Caenorhabditis elegans. Meanwhile, the resistance to toxicants was increased in long-lived animals. Here, we show that farnesoid X receptor (FXR) agonist obeticholic acid (OCA), a marketed drug for the treatment of cholestasis, may extend the lifespan and healthspan both in C. elegans and chemical-induced early senescent mice. Furthermore, OCA increased the resistance of worms to toxicants and activated the expression of detoxification genes in both mice and C. elegans. The longevity effects of OCA were attenuated in Fxr ^-/- mice and Fxr homologous nhr-8 and daf-12 mutant C. elegans. In addition, metabolome analysis revealed that OCA increased the endogenous agonist levels of the pregnane X receptor (PXR), a major nuclear receptor for detoxification regulation, in the liver of mice. Together, our findings suggest that OCA has the potential to lengthen lifespan and healthspan by activating nuclear receptor-mediated detoxification functions, thus, targeting FXR may offer to promote longevity.

Interfering hepatitis B virus (HBV) capsid assembly holds promise as a therapeutic approach for chronic hepatitis B (CHB). Novel anti-HBV agents are urgently needed to overcome drug resistance challenges, with targeted protein degradation (TPD) emerging as a hopeful strategy. Herein, we report the first degradation of HBV core protein (HBC), a multifunctional structural protein, using small-molecule degraders developed by hydrophobic tagging (HyT) technology. Structure-activity relationship (SAR) analysis identified compound HyT-S7, featuring an adamantyl group, exhibiting potent inhibitory activity (EC₅₀ = 0.46 μmol/L, HepAD38 cells) and degradation ability (DC₅₀ = 3.02 ± 0.54 μmol/L) in a dose- and time-dependent manner. Mechanistic studies demonstrated that the autophagy-lysosome pathway was a potential driver of HyT-S7-induced HBC degradation. Remarkably, HyT-S7 effectively degraded 11 drug-resistant mutants, including highly resistant strains P25G and T33N, to Phase III drug GLS4. Furthermore, cellular thermal shift assay, surface plasmon resonance assay, and molecular dynamics simulations revealed the precise mode of HyT-S7 binding to HBC with the adamantyl group potentially mimicking protein misfolding to facilitate HBC degradation. This first proof-of-concept study highlights the potential of HyT-mediated TPD in HBC as a promising avenue for discovering novel HBV and other antiviral agents with favorable drug resistance profiles.

OBJECTIVES: To investigate the inhibitory effect of Fuzheng Huaji Decoction against non-small cell lung cancer (NSCLC) cells in vitro and explore the underlying mechanism. METHODS: The active ingredients and targets of Fuzheng Huaji Decoction were identified using TCMSP and SwissTargetPrediction databases. NSCLC-related targets from GeneCards and PharmGKB were intersected with the targets of the Decoction, and a protein-protein interaction (PPI) network was constructed to identify the core targets, which were analyzed with GO and KEGG pathway enrichment analysis. Cultured A549 cells were treated with different concentrations of Fuzheng Huaji Decoction-medicated serum, and the changes in cell proliferation, apoptosis, and protein expressions were examined using CCK-8 assay, annexin V-FITC/PI staining and Western blotting. RESULTS: Fuzheng Huaji Decoction contained 140 active ingredients, and 707 drug-disease intersecting targets were identified. Among these targets, TP53, AKT1, HIF1A, GAPDH, ALB, EGFR, CTNNB1, and TNF were identified as the core targets which were involved in the biological processes related to kinases and receptors and the PI3K-AKT, Ras, calcium, and MAPK pathways. Molecular docking studies indicated strong binding affinity of the active ingredients with TP53, AKT1, and HIF1A. In cultured A549 cells, treatment with 2.5%, 5%, and 10% Fuzheng Huaji Decoction-medicated serum significantly inhibited cell proliferation, promoted cell apoptosis, and downregulated the expression levels of HIF1A, p-AKT (Thr308), and TP53 proteins. CONCLUSIONS Fuzheng Huaji Decoction inhibits proliferation of NSCLC cells possibly by downregulating the expressions of HIF1A, p-AKT (Thr308), and TP53.
Humans ; Carcinoma, Non-Small-Cell Lung/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Cell Proliferation/drug effects* ; Apoptosis/drug effects* ; Lung Neoplasms/metabolism* ; A549 Cells ; Proto-Oncogene Proteins c-akt/metabolism* ; Protein Interaction Maps ; Signal Transduction/drug effects* ; Cell Line, Tumor