搜索结果：Journal of cell science

Multiple myeloma (MM) typically evolves from monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM). Its progression is accompanied by significant tumor heterogeneity and immune microenvironment remodeling, and MM remains largely incurable despite therapeutic advances. Elucidating cellular heterogeneity and regulatory mechanisms involved in disease progression is critical for understanding MM pathogenesis. Single-cell RNA sequencing (scRNA-seq) provides important support for systematically elucidating the molecular mechanisms and immune regulatory pathways associated with the progression of MM. The scRNA-seq datasets from healthy donors (HD), MGUS, SMM, and MM patients were obtained from the GEO database to analyze plasma cell transcriptional heterogeneity within the bone marrow microenvironment. Plasma cell differentiation trajectories were inferred using pseudotime analysis, and intercellular communication and transcription factor regulatory networks were predicted using CellChat and SCENIC analyses. In addition, functional validation of ELK3 was performed in vitro using MM cell lines RPMI 8226 and U266. In this work, we used scRNA-seq to thoroughly evaluate the heterogeneity of MM. We identified a tumor-associated plasma cell subtype, C4 TTN+ plasma cell subtype, which was predominant in SMM and MM and was significantly enriched throughout the intermediate and final phases of differentiation. We hypothesized that C4 TTN+ plasma cell subtype may be associated with the progression of MM. We also found that the TGFb signaling pathway played a key role in the interaction between C4 TTN+ plasma cell subtype and various cell types within the TME. Furthermore, in vitro experiments validated the positive regulatory effects of ELK3 on plasma cell proliferation, migration, and cell viability in MM. In summary, this study, based on scRNA-seq analysis, identified a C4 TTN+ plasma cell subtype potentially associated with ELK3. This subtype may be associated with progression and immune evasion in MM, thereby providing a potential new direction for targeted immunotherapeutic strategies in MM.

The fact that cells die during development, metamorphosis and tissue homeostasis was recognized well over a century ago. However, aside from noting and classifying such cell death events, research into the mechanisms of regulated cell death did not 'take off' until about 35 years ago. Since then, our understanding of the different ways that cells die and how this comes about has blossomed. In celebration of the 100-year anniversary of the Journal of Cell Science's publisher The Company of Biologists, this Perspective article presents an overview of the past 100-plus years of cell death research, touching on the history of apoptosis, necroptosis and other forms of cell death.

Sickle cell disease (SCD) disproportionately affects racial and ethnic minority groups in the US and is associated with high levels of morbidity and health care utilization. However, population-level geographic differences and temporal variation in SCD hospitalization outcomes remain incompletely characterized. To assess temporal and regional patterns of SCD hospitalizations in New York State from 2009 through 2022. This retrospective cross-sectional study analyzed inpatient SCD hospitalizations recorded in the New York State Statewide Planning and Research Cooperative System deidentified database between January 1, 2009, and December 31, 2022. The analytic sample included 42 271 hospitalizations after exclusion of records with missing demographic, cost, or facility information. Data analysis was conducted from July 17, 2024, to February 14, 2025. Hospitalization with SCD across 8 state-defined health service areas. Outcomes included regional distribution of hospitalizations, mean length of stay, mean total charges, and trends in severity of illness and risk of mortality as defined by the All Patient Refined Diagnosis Related Groups classification system. Demographic and regional distributions were compared across years and regions. Among 42 271 SCD hospitalizations (21 777 female [51.5%]), most occurred among individuals identified as Black (35 318 [83.6%) compared with White (750 [1.8%]), multiracial (242 [0.6%]), and other race or ethnicity 5956 (14.1%) and were aged 18 to 29 (16 794 [39.7%]) or 30 to 49 years (13 480 [31.8%]). New York City accounted for the largest proportion of hospitalizations statewide. There were significant differences in the length of stay and total charges across service areas; Central New York had the longest mean (SD) length of stay of 6.3 (7.3) days, followed by the Hudson Valley (6.2 [7.2]) days, while Long Island had the highest mean (SD) total charges at $59 476.3 ($63 823.5). The proportion of hospitalizations classified as major severity increased from 751 of 5897 (12.7%) in 2009 to 1011 of 3709 (27.3%) in 2022, and the proportion classified as major risk of mortality increased from 170 of 5897 (2.9%) to 469 of 3709 (12.6%) during the same period. Long Island had the highest proportion of hospitalizations with major risk of mortality (93 of 970 [9.6%]), whereas New York City exhibited one of the lower proportions of major risk of mortality (1531 of 27 923 [5.5%]) despite high hospitalization volume. In this cross-sectional study, geographic and temporal differences in SCD hospitalization outcomes were observed across New York State during a 14-year period. These findings suggest the need for region-specific strategies to improve access to specialized care, reduce severe outcomes, and optimize health care resource use for individuals living with SCD.

Merkel cells are epithelial cells involved in the discrimination of light touch. Situated in the skin, these specialised cells decode mechanical cues through the mechanosensitive ion channel PIEZO2. Merkel cell carcinoma lines have been widely used as in vitro models for Merkel cells and like the native cell, they exhibit mechanically evoked currents. Herein, we show that unlike the native Merkel cell, that principally uses PIEZO2, mechanically evoked currents in the Merkel cell carcinoma line MCC13 are predominantly carried by PIEZO1, with variable contributions from PIEZO2. Slowly inactivating current types in these cells are carried by PIEZO1 complexed with auxiliary subunits MDFIC or MDFI. Moreover, PIEZO1 strongly influenced the endogenous levels of MDFIC, a known transcriptional repressor, whereby the loss of PIEZO1 dramatically reduced cellular levels of MDFIC. This suggests that the removal of PIEZO1 from a cell will likely have influences on the cellular transcriptome beyond its canonical Ca2+-based signalling pathways. In summary, utilizing MCC13 as a simple in vitro model of native Merkel cell mechanotransduction should be carried out with caution.

Circulating tumor cells (CTCs) serve as critical biomarkers for diagnosis, treatment monitoring, and prognosis evaluation of lung cancer. However, their trace abundance in blood makes the development of integrated technologies for CTCs isolation, enrichment, and detection with both high specificity and sensitivity a key bottleneck for clinical translation. This study proposed a novel cascade cell-assisted enhanced fluorescence strategy that integrates dual-targeting recognition, cell-mediated cascade assembly, and enzymatic amplification. This dual-targeting strategy achieves efficient capture and enrichment of human non-small cell lung cancer cell line A549 cells through EpCAM antibodies, and uses dual-aptamer probe-mediated EGFR recognition to further improve detection specificity. Subsequently, cascade assembly of acute lymphoblastic leukemia CCRF-CEM cells around target cells combined with enzyme-catalyzed signal amplification constructed a highly sensitive fluorescence-based detection system, with a linear range of 10-104 cells/mL, a limit of detection (LOD) as low as 10 cells/mL, and recovery rates of 94.25%-105.41%. Importantly, clinical validation using whole blood samples from lung adenocarcinoma patients (n = 30) and healthy donors (n = 23) showed significant differentiation (P < 0.05, AUC of 0.728), underscoring its diagnostic potential. Moreover, its modular expandability allows extension to other tumor CTCs detection by replacing target recognition units.

The innate immune signaling pathway cGAS-STING plays an important role in the recognition of cytosolic nucleic acids and the induction of the interferon-dependent antiviral response. Despite the significant research interest in this cascade in the context of immune system function, the mechanisms regulating cGAS-STING signaling and the switch between its pro-inflammatory and pro-apoptotic effects remain largely underexplored. According to publicly available RNA-seq data and microarray analyses, SETD7 lysine methyltransferase participates in interferon signaling in cancer cells. This study aims to elucidate the role of SETD7 in the regulation of the STING-dependent immune response in human lung adenocarcinoma (LUAD) cells. For this purpose, we developed a reproducible and cost-effective method for inducing the STING cascade by transfecting cells with salmon sperm DNA (sspDNA). We demonstrated that sspDNA efficiently induces phosphorylation of the key components of the STING-TBK1-IRF3 signaling pathway and activates the expression of interferons and pro-inflammatory cytokines. Using this approach, we further demonstrated that SETD7 is involved in the regulation of the IRF3-dependent transcriptional program. Suppression of SETD7 was associated with changes in the expression of genes related to innate immune response and apoptosis, including increased levels of IFNA1, IL1B, BAK1, BBC3 (PUMA), and BCL2. Furthermore, attenuation of SETD7 expression reduced the lentiviral transduction efficacy in H1299 cells. These results suggest that SETD7 may play a role in regulating the switch in STING signaling between pro-inflammatory and pro-apoptotic responses in LUAD cells.

Carbohydrate Response Element Binding Protein (ChREBP) is a transcription factor known to regulate glucose metabolism and other metabolic processes in various tissues, but its role in lung adenocarcinoma (LUAD) remains poorly understood. In this study, we investigated ChREBP expression and its role in regulating gene expression in LUAD cell lines. Using RT-qPCR, we assessed the expression of ChREBP-&#x3b1; and ChREBP-&#x3b2; isoforms in NCI-H1975, NCI-H1650, and NCI-H2228 LUAD cell lines. The NCI-H1975 cells exhibited the highest levels of both ChREBP isoforms, with a particularly pronounced expression of ChREBP-&#x3b2;. To explore the regulatory role of ChREBP, we generated NCI-H1975 cells with inducible expression of a dominant-negative mutant of human ChREBP (dnChREBP). Overexpression of dnChREBP led to a significant reduction in colony formation and impaired cell migration. Transcriptome analysis revealed 57 upregulated genes and 593 downregulated genes in dnChREBP-expressing cells compared to control cells. Functional annotation and gene set enrichment analysis revealed that the enriched genes were associated with cancer-related processes, including cell proliferation and epithelial-to-mesenchymal transition (EMT). Gene network analysis highlighted 17 downregulated hub genes, with 8 of these genes being associated with EMT. Interestingly, ChREBP and its transcriptionally regulated genes, including 4 top downregulated genes, 5 top upregulated genes, and 5 hub genes identified in NCI-H1975 cells overexpressing dnChREBP, showed significant prognostic value, as their expression levels correlated with overall survival in LUAD patients. Our findings suggest that ChREBP regulates distinct transcriptional programs in LUAD cells and ChREBP and its regulatory network may play a potential role in LUAD progression and patient outcomes.

Head and neck squamous cell carcinoma (HNSCC) is a highly heterogeneous malignancy with poor prognosis and limited predictive biomarkers for therapy response. Characterizing malignant cell heterogeneity may improve prognostic and therapeutic stratification. We integrated single-cell RNA sequencing (scRNA-seq) data from 58 HNSCC patients (181,003 cells) to define malignant cell subpopulations, their differentiation states, developmental trajectories, cell-cell interactions, and spatial localization. Coexpression gene modules and meta-programs were identified using hdWGCNA and NMF. These programs were projected onto bulk RNA-seq datasets to classify HNSCC subtypes and examine associations with clinical outcomes, tumor microenvironment (TME), genomic instability, and predicted response to immune checkpoint inhibitors (ICIs). Twelve malignant clusters were identified with distinct clinical and molecular features. MC-5 exhibited a stem-like phenotype associated with poor prognosis, while MC-7 and MC-11 showed high TME communication and immune engagement. Coexpression analysis revealed 16 modules and eight meta-programs encompassing proliferation, differentiation, stress response, and immune activity. Translation to bulk RNA-seq defined three HNSCC subtypes (MS-1, MS-2, MS-3) with divergent survival, immune infiltration, stromal composition, and genomic features. MS-2, an immune-enriched subtype, demonstrated superior survival, high HPV positivity, and predicted ICI responsiveness. A 25-gene malignant cell score (MCScore) robustly predicted both prognosis and immunotherapy response. This study provides a comprehensive map of malignant cell heterogeneity in HNSCC, identifies key functional expression programs, and defines molecular subtypes with clinical and therapeutic relevance. Malignant cell-specific signatures, such as MCScore, offer promising tools for patient stratification and precision immunotherapy.

Lung adenocarcinoma (LUAD) is a leading cause of cancer-related mortality worldwide. The tumor microenvironment (TME) plays a pivotal role in LUAD progression, but the specific molecular mechanisms driving malignancy and immune evasion remain incompletely understood. Bone marrow stromal cell antigen 2 (BST2) has been implicated in other cancers, yet its functional role and therapeutic potential in LUAD require further elucidation. We integrated single-cell RNA sequencing (scRNA-seq) data from primary LUAD tissues and normal lung tissues with bulk RNA-seq data from The Cancer Genome Atlas (TCGA)-LUAD and GEO cohorts. Malignant epithelial cells were identified using inferCNV analysis. Cellular trajectory and cell-cell communication analyses were systematically performed to delineate the malignant transformation process and its interactions with the TME. High-dimensional weighted gene coexpression network analysis further identified key functional modules within malignant subpopulations. The oncogenic role of BST2 was comprehensively validated through differential expression analysis, survival analysis, gene set enrichment analysis (GSEA), and in vitro cellular assays, including RT&#x2011;qPCR, MTT, colony formation, Transwell invasion, and wound healing experiments. In addition, we employed the tumor immune dysfunction and exclusion (TIDE) algorithm to evaluate its association with immunotherapy response and performed drug screening via the Clue.io platform to explore its therapeutic potential. scRNA-seq analysis revealed significant heterogeneity in the TME and identified two distinct subpopulations of malignant epithelial cells. Trajectory analysis uncovered a specific lineage (Lineage 4) driving the normal-to-malignant transition, while cell communication analysis highlighted interactions between malignant cells and tumor-associated macrophages (TAMs) mediated by MIF and APP signaling pathways. High-dimensional weighted gene coexpression network analysis (hdWGCNA) identified a coexpression gene module (Module 1) specifically enriched in malignant subpopulations. By intersecting with a set of immunoregulatory genes, BST2 was ultimately determined as a key candidate oncogene. BST2 was significantly upregulated in malignant epithelial cells and TAMs, and its high expression was closely associated with poor patient prognosis. GSEA demonstrated that high BST2 expression was linked to the activation of crucial oncogenic pathways, including oxidative phosphorylation, Kras signaling, and epithelial-mesenchymal transition (EMT). In vitro validation further confirmed that BST2 knockdown suppressed LUAD cell proliferation, migration, and invasion. Furthermore, elevated BST2 expression was associated with reduced efficacy of immune checkpoint blockade (ICB) therapy. Our study unveils BST2 as a critical oncogene in LUAD, promoting tumor progression and influencing the TME, particularly via TAM recruitment. BST2 expression predicts patient prognosis and immunotherapy response, positioning it as a promising biomarker and therapeutic target.

Lung cancer remains a major global public health challenge, with lung adenocarcinoma being the most prevalent histologic subtype. Rosuvastatin, a widely used lipid-lowering agent, has recently attracted attention for its potential antitumor properties. This study investigates the underlying mechanisms and therapeutic potential of rosuvastatin in lung cancer. The effects of rosuvastatin were evaluated in lung adenocarcinoma cell lines using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) incorporation, Transwell migration and invasion assays, wound-healing assays, and flow cytometry for apoptosis analysis. RNA sequencing identified cell-cycle signaling pathways as the primary targets of rosuvastatin. Analysis of survival curves and differential gene expression between tumor and adjacent non-tumor tissues using public databases, including the Human Protein Atlas, Gene Expression Profiling Interactive Analysis (GEPIA), and Tumor Immune Estimation Resource (TIMER), suggested that polo-like kinase 1 (PLK1) may be a key target mediating the antitumor effects of rosuvastatin. Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used to confirm the differential expression of PLK1 and related cell-cycle proteins in lung adenocarcinoma cells following treatment with different doses of rosuvastatin. Furthermore, rescue experiments with PLK1 knockdown were performed to verify its role in the mechanism of rosuvastatin. A subcutaneous mouse xenograft model was established in vivo to assess the antitumor activity of rosuvastatin via PLK1 inhibition. Rosuvastatin exerted significant antitumor effects against lung adenocarcinoma both in vitro and in vivo. Mechanistic studies indicated that its anticancer activity is mainly mediated by downregulating PLK1 expression. By suppressing PLK1 expression, rosuvastatin inhibited cancer cell proliferation, migration, and invasion. These findings support the potential of rosuvastatin as a therapeutic agent for lung cancer, although further studies are needed to confirm its clinical utility.

Alzheimer's disease (AD) is characterized by progressive memory decline and neuronal loss, driven primarily by dysregulated inflammatory signaling. Signal transducer and activator of transcription 6 (STAT6) is a member of the STAT family and has been implicated in the progression of various diseases. This study aimed to investigate whether inhibiting STAT6 and its downstream target, suppressor of cytokine signaling 2 (SOCS2), could mitigate A&#x3b2;1-42-induced neuronal injury in both cellular and animal models of AD. AD-like pathology was induced in Sprague-Dawley rats (n&#x2009;=&#x2009;10 per group) by intracerebral administration of amyloid &#x3b2;1-42 (A&#x3b2;1-42). Radial arm maze test, elevated plus maze test, and passive avoidance task were performed to estimate cognitive impairments of AD rats. H&E and TUNEL staining were performed to analyze pathological changes as well as neuron loss and apoptosis. HT-22 neuron cells were exposed to A&#x3b2;1-42 to establish an AD-relevant in vitro model. Cell viability and apoptosis were evaluated via CCK-8 and flow cytometry. Bax and Bcl-2 levels were estimated by Western blotting. Reactive oxygen species (ROS) production was measured using the DCFH-DA assay. ChIP and luciferase reporter assays were performed to assess the interaction between STAT6 and SOCS2 promoter. The results showed that&#xa0;STAT6 expression was significantly upregulated in the hippocampus of rats following A&#x3b2;1-42 infusion. Silencing of STAT6 alleviated A&#x3b2;1-42-induced cognitive impairments in AD rats by significantly reducing neuronal apoptosis (as evidenced by fewer TUNEL-positive cells), attenuating oxidative stress (indicated by reduced MDA levels and restored activities of antioxidant enzymes SOD and GSH-Px), and downregulating SOCS2 levels in vivo. Consistently, in vitro experiments demonstrated that STAT6 knockdown in HT-22 hippocampal neurons markedly inhibited A&#x3b2;1-42-triggered apoptosis, intracellular ROS accumulation, and oxidative stress marker dysregulation, whereas overexpressed STAT6 exerted an opposite role. Notably, STAT6 was found to bind to the SOCS2 promoter region. Functionally, SOCS2 overexpression alone accelerated HT-22 cell injury, and critically, it abolished the neuroprotective effects conferred by STAT6 depletion, specifically reversing the suppression of apoptosis and oxidative stress. These findings implicate that the STAT6-SOCS2 axis contributes to AD pathology, and their inhibition exerts neuroprotective effects.

Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease for which novel therapeutic approaches are urgently needed. Transforming Growth Factor-&#x3b2; (TGF-&#x3b2;) plays a central role in IPF pathogenesis by activating lung fibroblasts. Inhibitor of DNA binding (ID) proteins are regulated by TGF-&#x3b2;; however, their role in IPF remains poorly understood. We aimed to evaluate the regulation of ID proteins in IPF and to determine their functional role in human lung fibroblasts (HLF) in vitro and pulmonary fibrosis in vivo. ID protein expression was assessed in lungs and lung fibroblasts from mice and patients with pulmonary fibrosis. In vitro, the effects of ID1/ID3 inhibition and overexpression on HLF proliferation, migration and differentiation into myofibroblasts were evaluated. Genetic and pharmacological approaches were used in vivo to determine the role of ID1/ID3 in pulmonary fibrosis. ID1/ID3 levels were elevated in lungs and lung fibroblasts from mice and patients with pulmonary fibrosis, as well as in HLFs treated with TGF-&#x3b2;. ID1/ID3 knockdown reduced proliferation, migration and differentiation of healthy and IPF-derived HLFs. Bleomycin-exposed ID1/ID3 double KO mice exhibited improved lung function and reduced fibrosis compared with WT mice. Pharmacological inhibition of ID1/ID3 decreased HLF proliferation, migration and differentiation in vitro and attenuated pulmonary fibrosis in vivo. Lung-specific inhibition of ID1/ID3 using adeno-associated viruses expressing short hairpins targeting ID1 and ID3 also reversed pulmonary fibrosis in mice. Mechanistically, ID1/ID3 inhibition reduced fibroblast proliferation through regulation of cell cycle genes and attenuated fibroblast differentiation via the MEK/ERK pathway. Simultaneous inhibition of ID1 and ID3 attenuates pulmonary fibrosis. Targeting ID1/ID3 represents a potential novel therapeutic strategy for IPF.

Sepsis is prevalent and life-threatening condition that is often associated with high mortality rates. Early stratification is significantly associated with prognosis. Traditional biomarkers show low specificity, while scoring systems are time-consuming. This study aimed to evaluate the predictive value of a novel biomarker (N-myc and STAT interactor (NMI)) in assessing disease severity and clinical outcome in sepsis. A total of 399 adult patients diagnosed with sepsis were recruited. The least absolute shrinkage and selection operator (LASSO), logistic regression analysis, and receiver operating characteristic curve (ROC) were used to prove the predictive value of NMI in serum for disease severity (septic shock) and clinical outcome (30-day mortality) in the training set (n = 302). Internal validation (n = 97) was utilized to guarantee the stability of the findings. The study revealed that NMI was independently associated with 30-day mortality and the occurrence of septic shock through LASSO and logistic analysis. The NMI concentrations of patients with septic shock (166.1 [98.8, 437.5]) and non-survivors (208.7 [113.5, 809.6]) were higher than those of sepsis patients (54.2 [45.4, 64.8]) and survivors (59.3 [48.0, 90.2]), respectively. The area under the curve (AUC) of NMI for predicting mortality and septic shock was 0.86 and 0.92, respectively, which significantly outperformed other biomarkers and scoring systems. The AUCs of new scoring systems containing NMI were all remarkably higher than the original scoring systems (APACHE II, SOFA, qSOFA) for the prediction of clinical outcome and disease severity. Stratification of NMI concentrations in Kaplan-Meier survival curves proved the 30-day survival rate decreased with the increasing level of NMI (P < 0.001). Serum NMI can serve as a novel parameter to predict the severity and clinical outcome of sepsis, potentially facilitating timely disease stratification and providing certain guidance for clinical decision-making.

Glucocorticoid (GC) steroid hormones mediate the stress response, which requires dynamic communication between brain regions to enable an organism to react to and overcome perceived stressors. While many effects of GCs on the brain have been well-studied, little is known about whether and how GCs regulate brain cell-cell communication through the release of small extracellular vesicles (sEVs), nanoparticles that carry bioactive molecules between cells. Here, we use total internal reflection fluorescence (TIRF) microscopy and the pH-sensitive sEV marker mCh-CD63-pHluorin to visualize sEV release in a neuronal cell line. We find that GCs stimulate sEV secretion and that this process requires the GTPase Rab27a and the enzyme neutral sphingomyelinase 2 (nSMase2), which catalyzes ceramide production and drives sEV formation. We further show that GCs promote sEV release by activating nSMase2 downstream of mitochondrial reactive oxygen species production and opening of the mitochondrial permeability transition pore (mPTP). These findings link GC impacts on mitochondria, specifically via mPTP opening, to nSMase2 activation and enhanced sEV release by neuronal cells.

Interstitial cells of Cajal (ICCs) are essential for maintaining normal intestinal function, and their dysfunction is closely associated with various gastrointestinal diseases. However, the developmental process of ICCs and their role in intestinal development remain unclear. In this study, we analyzed single-cell transcriptome data and found that odd-skipped-related 1 (OSR1) is expressed in the intestinal tissues of humans and zebrafish. Additionally, OSR1 expression was reduced in tissue biopsies obtained from IBD patients. To further explore the function of osr1 in intestinal development and disease, we generated transgenic lines Tg(osr1:mCherry) and Tg(osr1:eGFP). Furthermore, we found that these transgenic fish could label interstitial cells of Cajal-like cells (ICC-like cells) from 48&#x2009;h post-fertilization (hpf) to adulthood. osr1-positive ICC-like cells form a network structure that envelops the intestine and coordinates its development. Moreover, mutations in osr1 lead to developmental defects in the intestine, characterized by disrupted ICC-like cells network structure, intestinal wall hypoplasia, and increased intestinal permeability, potentially mediated through the regulation of yap1. Overall, our findings indicate that osr1 acts as a novel marker gene for ICC-like cells and plays a crucial role in intestinal development.

Cells possess intricate metabolic networks comprised of hundreds of enzymes. Despite extensive research, many of these enzymes remain uncharacterized. Identifying the function of these enzymes is crucial for advancing our understanding of cellular metabolism. However, multiple enzymes are not active in standard conditions, making them challenging to study. To overcome this challenge, we created a pipeline to track the upregulation of enzymes at the protein level during diverse growth conditions, suggesting a requirement for their activity in these conditions. To do this, we assembled a collection of &#x223c;180 of yeast strains, each containing an uncharacterized putative enzyme fused to a fluorophore and under the regulation of its own promoter. By subjecting the collection to 42 diverse environments, we identified the biologically relevant conditions for the upregulation of 16 proteins. We focused on one such putative alcohol dehydrogenase, Bdh2, whose expression was upregulated during nutrient-limited conditions, and functionally characterized it. More broadly, our discovery pipeline lays the foundation for uncovering new stress-induced enzymes. This has implications in the cell biology of metabolism and biotechnology.

According to traditional Chinese medicine theory, acute myocardial infarction (AMI) is primarily associated with qi stagnation and blood stasis. Simiao Yong'an (SMYA) decoction is a well-known prescription that clears heat, detoxifies, and promotes blood circulation. SMYA has been used in the treatment of ischemic heart diseases (IHD). However, further analysis is required to clarify the specific mechanisms through which SMYA improves AMI and to determine its therapeutic effects at different time points during the acute phase of myocardial infarction. This study is aimed at investigating the protective effects of SMYA against AMI at various time points and to explore its underlying mechanisms. The active ingredients in SMYA were identified through ultraperformance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). An integrated in silico approach was employed to predict potential targets of these compounds, and target-pathway associations were established by aligning the data with relevant databases. A cardiac ischemia/reperfusion (I/R) model in rats was created by ligating the left coronary artery, inducing ischemia for 45&#x2009;min, and allowing for 24&#x2009;h of reperfusion. SMYA treatment was administered for 7 days. Cardiac function was evaluated at different time points during the acute phase of myocardial infarction using echocardiography. Serum biochemical indexes were measured using a biochemical kit, and western blotting (WB) was used to analyze AKT, p-AKT, PI3K, p-PI3K, BAX, Bcl-2, and caspase-3 proteins. UPLC-Q-TOF/MS identified 25 components in SMYA, which were considered potential effective ingredients. Network analysis identified 161 key targets and 167 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways associated with SMYA, with the PI3K-Akt pathway being notably prominent. Experimental validation demonstrated that SMYA significantly reduced the levels of creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) in serum and improved left ventricular ejection fraction (LVEF) and fractional shortening (FS) after myocardial I/R injury in rats. Additionally, SMYA reduced myocardial cell apoptosis and activated the PI3K-AKT pathway in a dose-dependent manner. Molecular docking confirmed binding between SMYA components and AKT/BCL-2. This study elucidates the mechanisms underlying AMI and the molecular action of SMYA. SMYA alleviates I/R-induced AMI in rats by activating the PI3K-AKT pathway, suggesting its potential as a therapeutic target for myocardial remodeling. The dose- and time-dependent protective effects of SMYA suggest that the PI3K-AKT pathway and its downstream target BCL-2 constitute promising therapeutic targets for novel interventions in AMI.

This article presents a case of recurrent autoimmune hemolytic anemia in a child with a gain-of-function (GOF) mutation of the TLR7. This patient's condition contrasts with the six previously documented cases of GOF mutations in the TLR7, thereby expanding the phenotypic spectrum of such mutations and enhancing clinical comprehension of childhood systemic lupus erythematosus (cSLE). The article discusses the mechanisms by which TLR7 GOF mutations can result in autoimmune hemolytic anemia, explores the influence of cytomegalovirus (CMV) infection on the disease's development and progression, and emphasizes the therapeutic potential of hematopoietic stem cell transplantation for cases of TLR7 GOF mutations.

Mixed invasive mucinous adenocarcinoma and non-mucinous adenocarcinoma (mixed IMA/NMA) is a rare subtype of lung adenocarcinoma (LUAD) with limited available data. This study aimed to comprehensively analyze the characteristics of this rare entity. A total of 738 surgical cases were enrolled, including 349 pure invasive mucinous adenocarcinoma (IMA), 61 mixed IMA/NMA and 328 pure non-mucinous adenocarcinoma (NMA) cases. Using amplification refractory mutation system, immunohistochemistry, and DNA-/RNA-based next-generation sequencing (DNA and RNA NGS), distinct molecular features were identified in mixed IMA/NMA cases compared with IMA and NMA cases, particularly in EGFR, KRAS, and ALK alterations and PD-L1 expression status. Paired analysis of the IMA and NMA components within mixed IMA/NMA cases using DNA and RNA NGS revealed one to three shared genomic alterations between the two components in the same tumor. However, significant differences were observed in the levels of cancer-associated fibroblasts, protumor cytokines, MHC-II, coactivation molecules, T cells, and effector cells between the components. Similarly, multiplex immunofluorescence assay demonstrated that immune cell infiltration, including CD4+ and CD8+ T cells, was significantly higher in the NMA components compared to the IMA components. Postoperative follow-up revealed no significant difference in disease-free survival (DFS) or overall survival (OS) between NMA and mixed IMA/NMA cases; however, both groups showed significantly shorter DFS (P&#x2009;=&#x2009;0.011) and OS (P&#x2009;=&#x2009;0.027) compared to IMA cases. Together, this study provides a comprehensive characterization of the molecular profiles, clonal relatedness, tumor heterogeneity, and surgical outcomes of mixed IMA/NMA, which may inform diagnostic and therapeutic strategies for this rare LUAD subtype.