Antifungal heteroresistance has emerged as a clinical challenge across diverse species. In Candida glabrata, however, it has rarely been reported. Here, we characterized heteroresistance in a multicenter collection of 156 C. glabrata isolates, revealing a 25% prevalence with caspofungin specificity and 37°C dependent phenotypes. Transcriptomic profiling of the heteroresistant subpopulation under escalating drug pressure revealed an adaptive program centered on cell cycle and cell wall integrity, including several members of the calcineurin pathway. Mechanically, we identified that the phenotype was abolished by pharmacological inhibition or Δcnb1, Δcrz1 genetic deletion, confirming the regulatory role of calcineurin. Crucially, heteroresistance functioned as a reservoir for resistance, with in vitro descendant strains spanning a spectrum of MIC alterations. Mechanistically, we found the heteroresistance phenotype mostly independent of aneuploidy. To decipher the genetic basis, we applied an unbiased machine-learning framework to genomic data, which not only identified the canonical FKS2_F659del mutation but also prioritized novel candidate PIR2_G149_I167del, demonstrating its power to uncover drivers of resistance from complex datasets. In summary, our study established a stepwise model of heteroresistance in C. glabrata, wherein calcineurin serves as a master regulator that promotes a resistance reservoir, revealing a potential vulnerability that could be exploited to prevent treatment failure.

G-quadruplexes (G4s) are non-canonical DNA secondary structures that act as local replication barriers and transcriptional regulators. Whether G4 can simultaneously influence splicing, DNA replication, and long-range, trans-chromosomal gene regulation remains untested. Here we combined in vitro biophysics, CRISPR mutagenesis and multi-omics to dissect a conserved G4 motif (QS1) located ~173 bp upstream of the PAX9 transcription start site. CD spectroscopy confirmed that the wild-type, but not the G-to-T mutant sequence, folds into a stable parallel G4 under physiological K+. In human cells, disruption of the QS1 G4 changed chromatin accessibility, remotely down-regulated a cohort of X-linked genes, accelerated migration and delayed G1/S progression. Integrative analysis of ATAC-seq, RNA-seq profiling reveals that the QS1 G4 acts as a three-dimensional genome scaffold linking craniofacial gene dosage to cell-cycle and metabolic networks. Our findings establish a pleiotropic role for a single promoter G4 in coordinating DNA replication stress, chromatin architecture and trans-chromosomal transcriptional control.

Ovarian clear cell carcinoma (OCCC) is a highly aggressive gynecological malignancy characterized by distinct clinicopathological features and resistance to chemotherapy. Despite advances in multi-omics characterization, the translational regulatory landscape of OCCC remains unexplored. Here, we performed ribosome profiling to systematically investigate translation control mechanisms in OCCC. We conducted an integrated analysis of transcriptomic and translatomic data from 22 clinical specimens. This study is the first to analyze translational dysregulation in OCCC at sub-codon resolution using translational group data resources. Integrated analysis identified novel unannotated open reading frames (ORFs) encoding functional micropeptides. Furthermore, we uncovered widespread translational dysregulation in OCCC, with experimental validation confirming the pro-tumorigenic role of translationally upregulated RBM4. This study bridges a critical gap between genomic, transcriptomic, and proteomic landscapes in OCCC, offering valuable mechanistic insights into its pathogenesis.

BACKGROUND:Insulinoma is the most common functional pancreatic neuroendocrine tumors, typically originating from pancreatic β cells. However, the transcriptional heterogeneity and the regulatory role of transcription factor YY1 remain incompletely understood. METHODS:High-throughput bulk-RNA sequencing was conducted on insulinoma tumor samples. Unsupervised clustering algorithms were employed to identify molecular subtypes of insulinomas. YY1 mutation status was analyzed by whole-exome sequencing. Functional roles of YY1 were analyzed by Yy1-overexpression and knockdown in INS-1 cell line. Spatial transcriptomic characteristics of insulinoma were analyzed using Visium HD platform. RESULTS:Transcriptomic analysis identified two distinct expression patterns: a low-endocrine subtype and an endocrine subtype. The low-endocrine subtype was characterized by reduced PDX1 and insulin synthesis pathways, with an enrichment in exocrine-related functions. However, these two subtypes exhibited similar clinicopathological features. Notably, while YY1 mutation rates were comparable between the two subtypes, YY1 expression levels were significantly reduced in the low-endocrine subtype. Functional experiments demonstrated that Yy1 levels directly correlated with insulin production, as Yy1 overexpression in INS-1 cells markedly upregulated insulin processing genes and secretory pathways. Yy1 knockdown led to suppression of insulin. Furthermore, spatial transcriptomics confirmed that low-endocrine subtype possessed reduced insulin scores compared to the endocrine subtype. CONCLUSION:We found a low-endocrine subtype insulinoma with reduced YY1 expression and insulin synthesis transcriptomic signature. This highlights the transcriptomic heterogeneity of insulinomas.

OBJECTIVE:Arsenic trioxide (ATO) is a cornerstone of acute promyelocytic leukemia (APL) therapy but induces severe gut microbiota dysbiosis, limiting its efficacy and safety. This study investigated whether adjunctive Bifidobacterium pseudolongum (BP) could mitigate these adverse effects and enhance therapeutic outcomes. METHODS:16S rRNA gene sequencing data of gut microbiota were obtained from a cohort of 22 APL patients treated with ATO-based regimens (20 of 22 data were obtained and analysis further), accessible under BioProject ID PRJNA935705. To evaluate the within-sample microbial community richness and evenness, alpha and beta diversity indices were calculated. Using a murine APL model, we compared ATO monotherapy with ATO+BP co-treatment. Analyses included fecal metagenomic sequencing, single-cell RNA sequencing (sc-RNA-seq), flow cytometric immune profiling, and assessment of intestinal tight junction proteins (claudin-1, occludin, and ZO-1) via immunofluorescence. RESULTS:ATO treatment significantly reduced gut microbial diversity and depleted beneficial taxa. Sc-RNA-seq data showed that ATO could orchestrate the APL immune microenvironment mainly through functional activation of CD8+ T cells and monocytes. BP supplementation restored microbial homeostasis and synergistically enhanced ATO's antileukemic effect, reducing the leukemic burden in peripheral blood by 72% and in bone marrow by 64% compared to ATO alone. Mechanistically, BP preserved intestinal barrier integrity by upregulating tight junction protein expression and modulated anti-tumor immunity, notably increasing bone marrow CD8+ T cells by 2.21-fold. CONCLUSIONS:BP is an effective adjunct to ATO therapy, counteracting gut dysbiosis, intestinal damage, and the immune microenvironment while synergistically improving antileukemic efficacy. Targeting the gut-leukemia axis with BP represents a promising strategy for improving the precision and safety of APL treatment.

Background The overall survival rate of acute myeloid leukemia (AML) remains less than 30%. Metabolic reprogramming of leukemia cells, such as the Warburg effect, enables them to adapt to the microenvironment and thereby develop. Elucidating the landscape of lactate regulation in AML helps clarify the pathogenesis from the perspective of metabolic reprogramming and identify possibilities for optimizing current treatment modalities.Methods RNA and single-cell sequencing data for AML were obtained from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. Seurat, limma package algorithm and Weighted gene coexpression network analysis (WGCNA) were conducted to identify candidate lactylation-related genes (LRGs). Enrichment analyses and protein-to-protein interactions were used to clarify the functions. Univariate COX regression and machine learning algorithms (LASSO-logistic, SVM-RFE and Boruta) narrowed the range of LRGs.The DALEX package employed four machine learning models for validation. CIBERSORT analyzed the relationship between immune cell infiltration and key LRGs, while single-gene GSEA was utilized to evaluate the functions of LRGs. We evaluated the associations between hub LRGs and AML using a two-sample Mendelian randomization (MR) analysis. Molecular docking was used to screen for feasible drugs targeting the hub genes. Western blotting was performed to assess pan-lactylation levels in AML cell lines. qRT-PCR and immunohistochemistry were performed to detect GZMB/LSP1 expression in AML patients.Results Seven hub LRGs were identified in the AML groups: LSP1, MPO, GZMB, SPINK2, HLA-DRB1, HLA-DRA and POU2F2, of which GZMB and LSP1 passed MR test. The seven hub genes were enriched in immune and inflammatory pathways. GLM ultimately emerged as the optimal model validated by GEO datasets. Compared with healthy controls, Kasumi-1 cells exhibited elevated lactylation levels, with exogenous lactate treatment further increasing lactylation levels, whereas sodium oxamate administration had the opposite effect. Exogenous lactate treatment significantly upregulated the mRNA expression of GZMB and LSP1. (-)-Gallocatechin gallate and indomethacin bound well to GZMB, while benzo(a)pyrene and benzo(e)pyrene had good binding potential with LSP1.Conclusions We established lactylation as a critical regulator of AML, and GZMB and LSP1 were identified as lactylation-related clinical modeling indicators, which provides a foundation for choosing prognostic and therapeutic strategies for AML.

Hsa_circ_0002111 is highly expressed in papillary thyroid carcinoma (PTC). We therefore investigated its role and underlying mechanism in promoting PTC progression. Q-RT-PCR, dual-luciferase reporter assays, AGO2-RIP, CCK-8, colony formation, wound healing, and Transwell assays were employed to dissect the circRNA/miRNA/mRNA axis interactions and their functional phenotypes. Results showed that silencing hsa_circ_0002111 significantly inhibited the viability, migration, and invasion of KTC-1 cells. Mechanistic studies demonstrated that hsa_circ_0002111 acted as a sponge for miR-432-5p, which in turn targeted the 3’UTR of CDKN2B and suppressed its expression. Functional assays revealed that either hsa_circ_0002111 knockdown or miR-432-5p overexpression inhibited migration and invasion, whereas inhibition of miR-432-5p reversed these cellular processes. Consistently, siRNA-mediated knockdown of CDKN2B phenocopied the motility defects observed upon hsa_circ_0002111 silencing, suggesting that CDKN2B is a functionally relevant downstream effector. Thus, the hsa_circ_0002111/miR-432-5p/CDKN2B axis is implicated in papillary thyroid carcinoma progression. These findings revealed a novel regulatory mechanism of hsa_circ_0002111 in KTC-1 cells and suggest its potential as a therapeutic target, offering new insights for future clinical applications and research directions in thyroid cancer.

Primary cutaneous T-cell lymphoma (CTCL) comprises a group of rare, aggressive non-Hodgkin lymphomas, of which mycosis fungoides (MF) and Sézary syndrome are the most common subtypes. In the absence of a universally accepted standard of care for advanced stages, allogeneic hematopoietic stem cell transplantation (allo-HSCT) offers curative potential; however, post-transplant relapses remains the principle cause of treatment failure, making effective maintenance strategies crucial. This report describes the case of a young woman with stage IVA MF who, following failure of multiple conventional therapies, underwent haploidentical allo-HSCT and initiated chidamide maintenance upon achieving complete remission. This approach successfully consolidated remission for 10 months; however, the patient relapsed at 11 months post-transplant and ultimately died of neutropenic septic shock. This case suggests that post-transplant chidamide maintenance may have value in delaying disease progression in advanced MF; however, treatment-related toxicities and the risk of relapse remain significant clinical challenges requiring further investigation.

Glioblastoma (GBM), a malignant brain tumor, is characterized by a high recurrence rate and poor overall survival. Therefore, understanding the causes of recurrence and identifying strategies for its prevention are crucial. To identify the characteristics of recurrent tumor samples and the molecular mechanisms of tumor progression, we conducted a longitudinal analysis of GBM using single-cell transcriptomics and clinical data from the Chinese Glioma Genome Atlas (CGGA). Compared to the primary tumor, recurrent GBM samples showed a more immunosuppressive environment. We identified an endothelial cell subpopulation, Edo4, that is characterized by high endothelial-to-mesenchymal transition (EMT) and is correlated with poor prognosis in recurrent tumors. It was regulated by the NR2F2 transcription factor specifically in the recurrent GBM samples. Cell-cell interaction analysis revealed that pericytes affect endothelial cell migration through the JAM2-(ITGA3+ITGB1) ligand-receptor axis, promoting GBM progression. Targeting the Edo4 cell cluster and the JAM2-(ITGA3+ITGB1) ligand-recep tor pair may be a promising strategy to prevent GBM recurrence.

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by the presence of the Philadelphia chromosome and the resulting fusion proteins with abnormal tyrosine kinase activity. The treatment of CML includes allogeneic hematopoietic stem cell transplantation (allo-HSCT) and TKI drug therapy. The BCR::ABL1 T315I mutation in CML leads to resistance to first- and second-generation tyrosine kinase inhibitors (TKIs), but is sensitive to two third-generation TKIs, olverembatinib and ponatinib. Olverembatinib, an oral third-generation BCR::ABL1 TKI, has preclinical activity against T315I-mutated CML. Here, we present a case of a CML patient with the T315I mutation who experienced late relapse with blast crisis 10 years after allo-HSCT. Following hematologic recovery and achievement of major molecular response (MMR) through induction therapy, the patient underwent a second allo-HSCT from his haploidentical sister. After successful engraftment, the patient received regular consolidation maintenance therapy with olverembatinib.

Aging leads to a gradual decline in immune function, termed immunosenescence, which significantly elevates the susceptibility to infections, cancers, and other aging-related diseases. Recent advancements have shed light on the molecular underpinnings of immune aging and pioneered novel therapeutic interventions to counteract its effects. Mesenchymal stem cells (MSCs)-a type of multipotent stromal cells with regenerative potential, low immunogenicity, and strong immunomodulatory properties-are increasingly recognized as a promising therapeutic option to reverse or alleviate immunosenescence-related dysfunction. This review systematically summarizes recent discoveries on how MSCs counteract immune aging, particularly their ability to rejuvenate aged immune cells and restore immune homeostasis. It also addresses key challenges, such as variations in MSC sources, donor variability, and the lack of standardized protocols, while proposing future directions to enhance therapeutic precision. Although preclinical and clinical studies highlight the potential of MSC-based strategies for delaying immunosenescence, critical issues remain unresolved, including long-term safety and efficacy, optimizing cell delivery systems, and elucidating context-specific mechanisms. Addressing these challenges will accelerate the development of MSC-based therapies to combat aging-associated immune decline.

BACKGROUND:Extramedullary acute myeloid leukemia (eAML) is a rare subtype of AML. The clinical features, molecular mechanisms and prognosis of eAML remain controversial. This study aimed to systematically analyze the differences in clinical and molecular characteristics between eAML patients and AML patients, and evaluate the impact of this disease subtype on survival outcomes. METHODS:We retrospectively included 96 patients with eAML and 144 patients with AML from our center between 2015 and 2024. RESULTS:eAML patients had a higher tumor burden than AML patients, with significantly elevated white blood cells (P < .001), platelets (P < .001), LDH (P < .001), peripheral blood blasts (P < .001) and bone marrow blasts (P = .005). In terms of molecular genetics, the eAML group was enriched for TET2, DNMT3A, ASXL1, PTPN11 and KMT2A mutations, while NPM1 and U2AF1 mutations were uncommon. The median overall survival (20.1 months vs. 38.8 months, P = .0021) and median relapse-free survival (7.6 months vs. 20.8 months, P = .00027) were significantly shorter for eAML patients. KRAS mutations and chromosomal abnormalities of t(9;11) were associated with poor prognosis. Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) could improve the prognosis of eAML patients. In subgroup analysis, we found that patients with multiple extramedullary involvements, synchronous eAML, skin infiltration, and soft tissue involvement had a worse prognosis, while patients with central nervous system (CNS) infiltration had a better prognosis. CONCLUSION:Our study demonstrates that patients with eAML subtype have a worse prognosis, unique molecular features and higher tumor burden. Allo-HSCT might be an effective way to improve prognosis. This study provides evidence critical for risk stratification and treatment optimization in eAML.

Extramedullary relapse of acute lymphoblastic leukemia (ALL) is usually associated with poor prognosis. Chimeric antigen receptor T cell (CAR-T cell) therapy followed by allogeneic hematopoietic stem cell transplantation is beneficial for relapsed/refractory (r/r) B cell acute lymphoblastic leukemia (B-ALL). Here, we report a B-ALL patient with extramedullary relapse involving several organs, including multiple lymph nodes and the breast, kidney, uterus and pancreas. After treatment with CAR-T cell therapy, positron emission tomography/computed tomography (PET/CT) revealed that she went into remission, with an almost undetectable tumor mass. She subsequently received unrelated cord blood transplantation (UCBT). Although she achieved minimal residual disease (MRD)-negative remission after UCBT for 5 months, she relapsed at the 6th month after UCBT. This patient achieved remission after subsequent interferon-α treatment for two weeks but eventually died of severe pneumonia. This case highlights the possibility of unusual relapse sites after chemotherapy and that regular biopsy of the mass is not sufficient to assess the scope and location of recurrence. PET/CT may be a useful tool to monitor the scope of extramedullary recurrence and follow-up remission. Further understanding of the pathology of extramedullary relapse is warranted to improve the management of such challenging presentations. This case suggests the efficacy of CAR-T cell therapy combined with UCBT in adult B-ALL patients with extramedullary relapse.

Heart disease stands as the foremost global cause of mortality. In rodents, the heart possesses the remarkable ability for cardiac regeneration within the first 7 days post-birth. Furthermore, the transition to an oxygen-rich environment and altered nutrient availability trigger a profound shift in cardiac energy metabolism immediately after birth. Lactylation, which translates metabolic adjustments into enduring gene expression patterns, has been recognized for its role in this process. However, its role in heart development has remained unexplored. In this study, we conduct an integrated study combining global proteomics, lactylome, and genome-wide RNA sequencing to elucidate the role of lactylation throughout postnatal heart development. Our findings demonstrate a remarkable increase in non-histone lactylation levels as early as 1 week (1 w) to 6 weeks (6 w) postpartum and remained elevated from 6 months (6 m) onwards. However, the histone lactylation showed the opposite trend. Additionally, we propose that histone 4 lysine 12 lactylation (H4K12la) acts as a pivotal upstream regulatory element in the early postnatal mouse heart, from 1 w to 6 w postpartum. Our findings strongly suggest a significant connection between lactylation and postnatal cardiac development and highlight its involvement in gene expression regulation, thus offering potential mechanisms for targeting heart diseases.

Genome sequencing (GS) refers to a technology that comprehensively and systematically detects the DNA sequences of an individual's nuclear and mitochondrial genomes. It aims to identify genetic variants and investigate their roles in human health and disease progression. As an emerging diagnostic tool, GS offers significant support for clinical diagnosis due to its high throughput, accuracy, and comprehensiveness. However, the complexity of data analysis and interpretation requires substantial professional expertise and experience, posing considerable challenges. When applying GS technology for molecular diagnosis of genetic diseases, ethical and technical issues related to clinical application arise, including informed consent, diagnostic data interpretation, and defining the scope and content of clinical reports. This expert consensus outlines the core workflow of clinical genome sequencing (cGS), clarifies its testing scope and technical limitations, and provides key steps for data quality control, analysis, annotation, and variant interpretation. It also addresses controversial issues related to report content and informed consent. This consensus aims to assist professionals in accurately understanding and appropriately utilizing clinical genome sequencing, thereby improving diagnostic accuracy for genetic diseases, enhancing the clinical utility of the technology, and advancing medical scientific research.

Minimal residual disease (MRD) monitoring has been demonstrated to important in predicting prognosis in acute myeloid leukemia (AML) receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT), but the ideal time point and method remain unclear. Our study compared the prognostic value of multiparameter flow cytometry (MFC)-based and WT1 expression-based MRD a month before allo-HSCT [HSCT(-1 m)] and after allo-HSCT [HSCT(+ 1 m)], as well as next generation sequencing (NGS)-based MRD at HSCT(-1 m), HSCT(+ 1 m), 3 and 6 months after allo-HSCT [HSCT(+ 3 m) and HSCT(+ 6 m)] among 47 AML patients undergoing allo-HSCT. The MRD status by all the methods at HSCT(-1 m) was proved as a superior indicator with prognostic significance for disease progression, compared to that at HSCT(+ 1 m). For the NGS-based MRD, HSCT(+ 6 m) seemed to be the optimal detection time point, as supported by the optimal prognostic discrimination capability and the relatively high sensitivity for disease progression prediction. Moreover, our data showed that each individual method had some limitations in predicting prognosis; however, pre-transplant MRD monitoring by the combination of MFC, WT1 and NGS could greatly increase the sensitivity (100%) of identifying disease progression and greatly improve prognostic stratification. Our study may provide insights into the optimal time point and methodology for MRD monitoring in AML following allo-HSCT.

MicroRNAs (miRNAs) are noncoding RNAs that regulate the expression of target genes after transcription, and play important roles in the differentiation of hematopoietic stem cells. Many miRNAs are related to the occurrence of acute promyelocytic leukemia (APL) and play roles in the treatment response. Recently, we demonstrated that several miRNAs are differentially expressed in patients with relapsed and newly diagnosed APL; for example, miR-140-3p is significantly downregulated in patients with relapsed APL. In this study, via a dual luciferase assay, we verified that one of the direct target genes of miR-140-3p is hepatocyte growth factor (HGF). After different lentiviruses were transfected into NB4 cells, and flow cytometry and proliferation assays confirmed that low expression of miR-140-3p inhibited the differentiation and apoptosis of NB4 cells and induced proliferation by promoting cell cycle progression. In summary, our findings suggest that hepatocyte growth factor is a target gene of miR-140-3p. Moreover, upregulation of miR-140-3p expression in APL cells inhibits cell proliferation, arrests cell cycle progression, and promotes apoptosis and cell differentiation. Monitoring the levels of miR-140-3p and HGF may predict the risk of disease recurrence, and interfering with the miR-140-3p / HGF pathway may have therapeutic potential for treating recurrent APL.

In China,an estimated 48.42-82.11 million individuals are affected by rare diseases.This estimate is derived from the Orpha-net database,based on the global population of 7.55 billion and China's population of 1.39 billion in 2017(source:National Bureau of Statistics of China)[1,2].Patients with rare diseases in China commonly experience substantial diagnostic difficulties,and the 2018 China Rare Disease Survey showed that 72.97％of patients experienced misdiagnosis and required an average of 4.30 years and visits to 2.97 hospitals to obtain a definitive diagnosis.More-over,14.3％of patients underwent a diagnostic journey lasting 10 years or more,with 8.8％visiting more than 10 hospitals during this period[3,4].