OBJECTIVE To focus on the classic NOD-like receptor protein 3 （NLRP3）/Caspase-1/gasdermin D （GSDMD） pyroptosis pathway and explore the mechanism by which Huatan qushi huoxue formula （HQHF） inhibits macrophage pyroptosis to ameliorate metabolic dysfunction-associated steatohepatitis （MASH）. METHODS RAW264.7 cells were divided into 5 groups： Control group （10% blank serum）， Model group [10% blank serum+5 μg/mL lipopolysaccharide （LPS）]， HQHF-L group （2.5% drug-containing serum+7.5% blank serum+5 μg/mL LPS）， HQHF-M group （5% drug-containing serum+5% blank serum+5 μg/mL LPS）， and HQHF-H group （10% drug-containing serum+5 μg/mL LPS）. After 24 h of routine culture post-administration， cells and supernatants were collected for assays. Cell morphology was observed via scanning electron microscopy and phase-contrast microscopy； localization and expression of gasdermin D-N （GSDMD-N） were observed by immunofluorescence. Interleukin-1β （IL-1β） and IL-18 contents in supernatants were detected by ELISA； mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD were measured using real-time PCR and Western blot. RESULTS Compared with the Control group， the Model group showed typical pyroptotic morphology （cell membrane bulging and pore formation）， increased aggregation and fluorescence intensity of GSDMD-N on the cell membrane （ P ＜0.05）， significantly increased the contents of IL-1β and IL-18 in cell supernatants （ P ＜0.05）， and significantly up-regulated mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD in cells （ P ＜0.05）. Compared with the Model group， the HQHF-L， HQHF-M and HQHF-H groups showed improved pyroptotic morphology， reduced membrane localization and significantly weakened fluorescence intensity of GSDMD-N （ P ＜0.05）， significantly decreased the contents of IL-1β and IL-18 in cell supernatants （ P ＜0.05）， and significantly down-regulated mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD in cells （ P ＜0.05）. CONCLUSIONS HQHF inhibits LPS-induced macrophage pyroptosis， and its mechanism of improving MASH may be associated with the suppression of the activation of the classical NLRP3/Caspase-1/GSDMD pyroptosis pathway.

Periodontal disease is a risk factor for many systemic diseases such as Alzheimer's disease and adverse pregnancy outcomes. Cleft palate (CP), the most common congenital craniofacial defect, has a multifaceted etiology influenced by complex genetic and environmental risk factors such as maternal bacterial or virus infection. A prior case-control study revealed a surprisingly strong association between maternal periodontal disease and CP in offspring. However, the precise relationship remains unclear. In this study, the relationship between maternal oral pathogen and CP in offspring was studied by sonicated P. gingivalis injected intravenously and orally into pregnant mice. We investigated an obvious increasing CP (12.5%) in sonicated P. gingivalis group which had inhibited osteogenesis in mesenchyme and blocked efferocytosis in epithelium. Then glycolysis and H4K12 lactylation (H4K12la) were detected to elevate in both mouse embryonic palatal mesenchyme (MEPM) cells and macrophages under P. gingivalis exposure which further promoted the transcription of metallopeptidase domain17 (ADAM17), subsequently mediated the shedding of transforming growth factor-beta receptor 1 (TGFBR1) in MEPM cells and mer tyrosine kinase (MerTK) in macrophages and resulted in the suppression of efferocytosis and osteogenesis in palate, eventually caused abnormalities in palate fusion and ossification. The abnormal efferocytosis also led to a predominance of M1 macrophages, which indirectly inhibited palatal osteogenesis via extracellular vesicles. Furthermore, pharmacological ADAM17 inhibition could ameliorate the abnormality of P. gingivalis-induced abnormal palate development. Therefore, our study extends the knowledge of how maternal oral pathogen affects fetal palate development and provides a novel perspective to understand the pathogenesis of CP.
Animals ; Female ; Porphyromonas gingivalis ; Pregnancy ; Mice ; Cleft Palate/etiology* ; Glycolysis

Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans ; Dental Cementum/injuries* ; Consensus ; Diagnosis, Differential ; Cone-Beam Computed Tomography ; Tooth Fractures/therapy*

Successful cloning through somatic cell nuclear transfer (SCNT) faces significant challenges due to epigenetic obstacles. Recent studies have highlighted the roles of H3K4me3 and H3K27me3 as potential contributors to these obstacles. However, the underlying mechanisms remain largely unclear. In this study, we generated genome-wide maps of H3K4me3 and H3K27me3 in mouse pre-implantation NT embryos. Our analysis revealed that aberrantly over-represented broad H3K4me3 domain and H3K27me3 signal lead to increased bivalent marks at gene promoters in NT embryos compared with naturally fertilized (NF) embryos at the 2-cell stage, which may link to relatively low levels of H3K36me3 in NT 2-cell embryos. Notably, the overexpression of Setd2, a H3K36me3 methyltransferase, successfully restored multiple epigenetic marks, including H3K36me3, H3K4me3, and H3K27me3. In addition, it reinstated the expression levels of ZGA-related genes by reestablishing H3K36me3 at gene body regions, which excluded H3K27me3 from bivalent promoters, ultimately improving cloning efficiency. These findings highlight the excessive bivalent state at gene promoters as a potent barrier and emphasize the removal of these barriers as a promising approach for achieving higher cloning efficiency.
Animals ; Mice ; Histone-Lysine N-Methyltransferase/biosynthesis* ; Histones/genetics* ; Nuclear Transfer Techniques ; Female ; Gene Expression Regulation, Developmental ; Promoter Regions, Genetic ; Epigenesis, Genetic ; Embryo, Mammalian/metabolism*

Objective To evaluate the promoting effect of continuous low-intensity pulsed ultrasound(LIPUS)combined with microbubble(MB)cavitation therapy(hereinafter referred to as ultrasound cavitation therapy)on microcirculatory perfusion in the ischemic hindlimbs of mice,and to explore the non-invasive therapeutic potential of this treatment for limb arterial ischemic injury.Methods A mouse model of left hindlimb ischemia was established,and the mice were randomly assigned to 4 groups(16 mice per group)according to different treatment methods:model group,ultrasound contrast microbubble group(MB group),LIPUS treatment group(LIPUS group),and ultrasound cavitation therapy group(LIPUS+MB group).Mice in the model group were injected with 0.1 mL of normal saline via the tail vein,those in the MB group were injected with 0.1 mL of MB via the tail vein,those in the LIPUS group were treated with LIPUS on the ischemic hindlimb after injection of 0.1 mL of normal saline via the tail vein,and those in the LIPUS+MB group were treated with LIPUS on the ischemic hindlimb after injection of 0.1 mL of MB via the tail vein;each group was injected once a day for a total of 7 d.On the 1st,4th and 7th days after treatment,the microcirculatory perfusion in the ischemic hindlimbs of mice was evaluated using contrast-enhanced ultrasound.The effects of different treatments on promoting microcirculatory perfusion in the ischemic hindlimbs of mice were assessed by combining hematoxylin-eosin(H-E)staining and CD31 immunohistochemical staining of the gastrocnemius muscle tissue in the hindlimbs.Results The left hindlimb ischemia model was successfully constructed,and all model mice showed obvious ischemic microcirculation perfusion disorders with good model stability.After the 7th day of treatment,the LIPUS+MB group showed a increase in microcirculation perfusion in the ischemic hindlimb,with the ratio of microvascular flow on the ischemic to non-ischemic sides higher than that of the LIPUS group([94.33±4.51]%vs[70.33±2.09]%,P＜0.05).H-E staining results showed that the LIPUS+MB group had more newly formed capillaries and myofibroblasts in the gastrocnemius muscle,with better muscle structure repair compared to the LIPUS group,while the model group and MB group showed muscle cell necrosis,disorganized arrangement of muscle bundles,and sparse capillaries.CD31 immunohistochemical analysis further confirmed that ultrasonic cavitation therapy significantly outperformed traditional LIPUS treatment in promoting microcirculation perfusion,microvascular neogenesis,and tissue repair in ischemic skeletal muscles(CD31 relative expression level 5.03±0.33 vs 3.57±0.21,P＜0.01).Conclusion Compared with single LIPUS treatment,continuous ultrasound cavitation therapy has a more significant effect on promoting microcirculation perfusion in the ischemic hindlimb of mice,which provides a new strategy for microcirculatory perfusion disorders in skeletal muscles of limbs caused by peripheral arterial ischemic diseases.

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APC^min/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.

Ultrasound-stimulated microbubble cavitation(USMC)is a cavitation phenomenon triggered by oscillating microbubbles in ultrasound field,and this effect has led to breakthroughs in improving the permeability of cell membranes,enhancing the sensitivity of tumor cells to chemotherapy or immunotherapy,and increasing the penetration of thrombolytic drugs into blood clots and vascular recanalization.In recent years,the cross-fertilization of ultrasound medicine and materials science have given rise to the study of ultrasound-responsive multifunctional nanobubble to enhance USMC treatment.This article reviews the application and research progress of USMC in improving tissue perfusion in ischemic diseases,tumor chemotherapy and immunotherapy,ultrasonic thrombolysis,and other treatments.

Recent studies have highlighted spatially resolved multi-omics technologies,including spatial genomics,transcriptomics,proteomics,and metabolomics,as powerful tools to decipher the spatial heterogeneity of the brain.Here,we focus on two major approaches in spatial transcriptomics(next-generation sequencing-based technologies and image-based technologies),and mass spectrometry imaging tech-nologies used in spatial proteomics and spatial metabolomics.Furthermore,we discuss their applications in neuroscience,including building the brain atlas,uncovering gene expression patterns of neurons for special behaviors,deciphering the molecular basis of neuronal communication,and providing a more comprehensive explanation of the molecular mechanisms underlying central nervous system disorders.However,further efforts are still needed toward the integrative application of multi-omics technologies,including the real-time spatial multi-omics analysis in living cells,the detailed gene profile in a whole-brain view,and the combination of functional verification.

Non-conventional yeasts such as Yarrowia lipolytica, Pichia pastoris, Kluyveromyces marxianus, Rhodosporidium toruloides and Hansenula polymorpha have proven to be efficient cell factories in producing a variety of natural products due to their wide substrate utilization spectrum, strong tolerance to environmental stresses and other merits. With the development of synthetic biology and gene editing technology, metabolic engineering tools and strategies for non-conventional yeasts are expanding. This review introduces the physiological characteristics, tool development and current application of several representative non-conventional yeasts, and summarizes the metabolic engineering strategies commonly used in the improvement of natural products biosynthesis. We also discuss the strengths and weaknesses of non-conventional yeasts as natural products cell factories at current stage, and prospects future research and development trends.
Yeasts/genetics* ; Yarrowia/metabolism* ; Gene Editing ; Metabolic Engineering

Objective:To identify the risk factors of postoperative atrial fibrillation (POAF) in critically ill lung surgery patients and establish a nomogram.Methods:213 critically ill lung surgery patients were collected in Beijing Chaoyang Hospital from January 2018 to December 2021. Logistic analysis was used to analyze the risk factors of POAF. A nomogram was developed based on the verified risk factors. Results:The independent risk factors associated with POAF was mitral regurgitation ( OR=4.270, 95% CI: 1.380-13.213, P=0.012), cedilanid ( OR=14.619, 95% CI: 2.913-73.373, P=0.001), catecholamine ( OR=3.244, 95% CI: 1.144-9.203, P=0.027), pericardiotomy( OR=6.079, 95% CI: 1.362-27.128, P=0.009), systematic lymph node dissection( OR=5.460, 95% CI: 1.770-16.846, P=0.003). Nomogram model showed the ROC was 0.801(95% CI: 0.721-0.881, P<0.001). Conclusion:The risk factors of POAF in critically ill lung surgery patients are mitral regurgitation, cedilanid, catecholamine, pericardiotomy and systematic lymph node dissection. The nomogram predicted POAF better than other scoring systems.