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Calcium-activated chloride channels (CaCCs) are essential for epithelial secretion, neuronal transmission, and smooth muscle function. Among the Anoctamin family, Anoctamin 1 (ANO1) and Anoctamin 2 (ANO2) are classical CaCCs proteins. ANO1 has been identified as a potential therapeutic target due to its involvement in diseases such as cancer and cystic fibrosis. However, current high-throughput screening (HTS) systems face limitations in achieving subtype-specific detection and optimizing screening strategies. Stable HTS cell models expressing ANO1 or ANO2 were constructed via lentiviral transduction in Fischer mouse thyroid (FRT) cells. The models were validated using flow cytometry, Reverse Transcription Polymerase Chain Reaction(RT-PCR), and YFP-H148Q/I152L-based iodide fluorescence quenching assays. Patch-clamp electrophysiology was employed to characterize ANO1 and ANO2 current properties. Although these electrophysiological features have been previously reported, their application in HTS workflows had not been systematically evaluated. ANO1 displayed notable current rundown under sustained stimulation with high Ca 2+ or agonist concentrations, whereas ANO2 maintained stable currents under identical conditions. Based on these findings, an optimized screening strategy was developed, incorporating agonist concentration gradients and the timing of inhibitor application. This approach improved the specificity and reliability of modulator detection. A robust and functionally validated cell-based HTS platform for CaCCs modulator discovery was established. By integrating the electrophysiological characteristics of ANO1 into the screening design, the optimized strategy enhances the accuracy of identifying selective ANO1 modulators. This work provides a methodological basis for future mechanism-driven screening of CaCCs-targeted compounds.

The Piezo1 channel, a mechanosensitive non-selective cation channel, plays a critical role in mediating calcium (Ca 2+ ) influx in response to mechanical stimuli, which is vital for numerous physiological functions. However, the specificity and potency of current Piezo1 modulators are limited, and the existing screening methodologies are not sufficiently designed for high-throughput screening (HTS). To address these challenges, we developed a customized cellular model aimed at enhancing HTS efficiency for identifying potent and selective Piezo1 modulators. We utilized a cellular model that incorporates anoctamin-1 (ANO1), a calcium-activated chloride channel, along with a green fluorescent protein mutant with ultra-high halide sensitivity (YFP-H148Q/I152L) to monitor intracellular ion concentrations through fluorescence. This model can accurately detect changes in intracellular iodine ion (I ⁻ ) concentration and rapidly and sensitively screen drugs targeting the Piezo1 channel, demonstrating exceptional performance in high-throughput screening (HTS). Our development of a pharmacological cell screening model specifically designed for Piezo1 provides a valuable tool for investigating Piezo1 modulators and their pathophysiological roles in vitro.

Transient receptor potential vanilloid 4 (TRPV4) is a widely expressed cation channel that plays an important role in many physiological and pathological processes. However, most TRPV4 drugs carry a risk of side effects. Moreover, existing screening methods are not suitable for the high-throughput screening (HTS) of drugs. In this study, a cell model and HTS method for targeting TRPV4 channel drugs were established based on a calcium-activated chloride channel protein 1 Anoctamin 1 (ANO1) and a double mutant (YFP-H148Q/I152L) of the yellow fluorescent protein (YFP). Patch-clamp experiments and fluorescence quenching kinetic experiments were used to verify that the model could sensitively detect changes in intracellular Ca2+ concentration. The functionality of the TRPV4 cell model was examined through temperature variations and different concentrations of TRPV4 modulators, and the performance of the model in HTS was also evaluated. The model was able to sensitively detect changes in the intracellular Ca2+ concentration and also excelled at screening TRPV4 drugs, and the model was more suitable for HTS. We successfully constructed a drug cell screening model targeting the TRPV4 channel, which provides a tool to study the pathophysiological functions of TRPV4 in vitro.

Cultural heritage (CH) relics are irreplaceable records of human civilization, encompassing diverse historical, technological, and artistic achievements. Extracting their structural and compositional information without affecting their physical integrity is a critical challenge. This review summarizes recent advances in non-destructive techniques (NDTs) for CH analysis and emphasizes the balance between the depth of analysis and conservation ethics. Techniques are broadly categorized into spectrum-based, X-ray-based, and digital-based methods. Spectroscopic techniques such as Fourier transform infrared (FTIR), Raman, and nuclear magnetic resonance (NMR) spectroscopy provide molecular-level insights into organic and inorganic components, often requiring minimal or no sampling. X-ray-based techniques, including conventional and spatially resolved XRD/XRF and total reflection XRF (TRXRF), provide powerful means for crystal and elemental analysis, including in situ pigment identification and trace material analysis. Digital-based methods include high-resolution imaging, three-dimensional modeling, data fusion, and AI-driven diagnosis to achieve the non-invasive visualization, monitoring, and virtual restoration of CH assets. This review highlights a methodology shift from traditional molecular-level detection to data-centric and AI-assisted diagnosis, reflecting the paradigm shift in heritage science.

Purpose Chronic myeloid leukemia (CML) is a myeloproliferative disease derived from hematopoietic stem cells (HSCs) that harbor Philadelphia chromosome (Ph chromosome). In clinic, leukemia stem cells (LSCs) in CML are insensitive to the treatment with tyrosine kinase inhibitors, and are responsible for disease relapse. However, the molecular mechanisms for maintaining LSCs survival remain elusive. Methods CML patient-derived cell lines and BCR-ABL-induced CML mouse model were used to explore the role of YBX1 in regulating the survival of CML LSCs. Bone marrow transduction and transplantation, and colony-forming unit assay were used to investigate LSC function. The underlying mechanism of how YBX1 regulates LSCs survival were assessed using flow cytometry, RNA sequencing, western blot, RNA decay assay, co-immunoprecipitation and RNA immunoprecipitation. Results Here we show that RNA-binding protein YBX1 plays an important role in regulating survival of CML LSCs. We find that YBX1 expression is significantly increased in CML cells, and confirm that YBX1 is required for maintaining survival of LSCs. Deletion of YBX1 impairs the propagation of CML through blocking cell proliferation and inducing apoptosis of LSCs. Mechanistically, we find that YBX1 regulates expression of apoptotic associated genes. YBX1 cooperates with RNA m 6 A reader IGF2BPs to stabilize YWHAZ transcript in an m 6 A-dependent manner, and loss of YBX1 decreases YWHAZ expression by accelerating mRNA decay. Restoration of YWHAZ efficiently rescues the defects of YBX1-deficient CML cells. Conclusion Our findings reveal a critical role of YBX1 in maintaining survival of CML LSCs, which provides a rationale for targeting YBX1 in CML treatment.

Acute myeloid leukemia (AML) is a highly aggressive hematologic malignancy that arises from leukemia stem cells (LSCs) transformed from normal hematopoietic stem progenitor cells (HSPCs). Compared to normal HSPCs, LSCs acquire various adaptive properties that enable their survival and regenerative capacity under environmental challenges. However, the key regulators underlying these adaptative features in AML LSCs remain largely unknown. Here, the mechanosensor PIEZO1 as a key factor in promoting acute myeloid leukemia progression and maintaining LSC stemness by HIF1A‐SLC7A11 axis‐mediated ferroptosis defense is uncovered. PIEZO1 deletion impairs cystine uptake and induces ROS and lipid peroxidation, causing ferroptosis of leukemia cells. Mechanistically, HIF1A mediates the function of PIEZO1 by transcriptionally regulating the expression of SLC7A11 and SLC3A2. Further, inhibition of PIEZO1 and SLC7A11 suppresses LSC function and contributes to AML treatment. In summary, this findings reveal that LSCs control cystine uptake through the PIEZO1‐HIF1A‐SLC7A11 axis to defend against ferroptosis, representing a unique vulnerability of LSCs.

Chronic myeloid leukemia (CML) is a myeloproliferative disease derived from hematopoietic stem cells (HSCs) that harbor Philadelphia chromosome (Ph chromosome). In clinic, leukemia stem cells (LSCs) in CML are insensitive to the treatment with tyrosine kinase inhibitors, and are responsible for disease relapse. However, the molecular mechanisms for maintaining LSCs survival remain elusive. CML patient-derived cell lines and BCR-ABL-induced CML mouse model were used to explore the role of YBX1 in regulating the survival of CML LSCs. Bone marrow transduction and transplantation, and colony-forming unit assay were used to investigate LSC function. The underlying mechanism of how YBX1 regulates LSCs survival were assessed using flow cytometry, RNA sequencing, western blot, RNA decay assay, co-immunoprecipitation and RNA immunoprecipitation. Here we show that RNA-binding protein YBX1 plays an important role in regulating survival of CML LSCs. We find that YBX1 expression is significantly increased in CML cells, and confirm that YBX1 is required for maintaining survival of LSCs. Deletion of YBX1 impairs the propagation of CML through blocking cell proliferation and inducing apoptosis of LSCs. Mechanistically, we find that YBX1 regulates expression of apoptotic associated genes. YBX1 cooperates with RNA m A reader IGF2BPs to stabilize YWHAZ transcript in an m A-dependent manner, and loss of YBX1 decreases YWHAZ expression by accelerating mRNA decay. Restoration of YWHAZ efficiently rescues the defects of YBX1-deficient CML cells. Our findings reveal a critical role of YBX1 in maintaining survival of CML LSCs, which provides a rationale for targeting YBX1 in CML treatment.

N 6 -methyladenosine (m 6 A) is a commonly modification of mammalian mRNAs and plays key roles in various cellular processes. Emerging evidence reveals the importance of RNA m 6 A modification in maintaining stem cell function in normal hematopoiesis and leukemogenesis. In this review, we first briefly summarize the latest advances in RNA m 6 A biology, and further highlight the roles of m 6 A writers, readers and erasers in normal hematopoiesis and acute myeloid leukemia. Moreover, we also discuss the mechanisms of these m 6 A modifiers in preserving the function of hematopoietic stem cells (HSCs) and leukemia stem cells (LSCs), as well as potential strategies for targeting m 6 A modification related pathways. Overall, we provide a comprehensive summary and our insights into the field of RNA m 6 A in normal hematopoiesis and leukemia pathogenesis.

A lack of standardization in heart team implementation potentially leads to suboptimal decision-making quality, and we previously established a modified heart team protocol to improve the decision-making quality. The present trial was to validate the effect of the modified heart team implementation protocol on improving the decision-making quality versus the conventional protocol in complex coronary artery disease (CAD). Eligible interventional cardiologists, cardiac surgeons and non-interventional cardiologists were randomly allocated to the intervention or control arm and established 12 heart teams in each arm. The 12 heart teams in each arm were randomly divided into 6 pairs, and 480 historic cases with complex CAD into 6 sets of 80 cases. In each arm, each set of 80 cases was discussed independently by one pair of heart teams, with each case finally receiving two heart team decisions ('pairwise decisions'). The intervention arm conducted heart team decision-making according to the previously established protocol and the control arm based on guideline recommendations. The primary outcome was the overall percent agreement of the inter-team pairwise decisions. Decision-making appropriateness was further analysed. A total of 36 cardiac surgeons, 36 interventional cardiologists and 12 non-interventional cardiologists from 26 centres were enrolled. The overall percent agreement was significantly higher in the intervention arm than the control arm (72.1% vs 65.8%, P = 0.04; kappa 0.51 vs 0.37). Both team-level (19.4% vs 33.0%; P < 0.001) and specialist-level (interventional cardiologists, 19.8% vs 37.7%, P < 0.001; cardiac surgeons, 19.8% vs 28.7%, P < 0.001) inappropriateness rate of decision-making was significantly lower in the intervention arm than the control arm. The modified heart team implementation protocol improved the decision-making quality and appropriateness compared with the guideline-based protocol.

Trustworthy multi-view classification (TMVC) addresses the challenge of achieving reliable decision-making in complex scenarios where multi-source information is heterogeneous, inconsistent, or even conflicting. Existing TMVC approaches predominantly rely on globally dense neighbor relationships to model intra-view dependencies, leading to high computational costs and an inability to directly ensure consistency across inter-view relationships. Furthermore, these methods typically aggregate evidence from different views through manually assigned weights, lacking guarantees that the learned multi-view neighbor structures are consistent within the class space, thus undermining the trustworthiness of classification outcomes. To overcome these limitations, we propose a novel TMVC framework that introduces prototypes to represent the neighbor structures of each view. By simplifying the learning of intra-view neighbor relations and enabling dynamic alignment of intra- and inter-view structure, our approach facilitates more efficient and consistent discovery of cross-view consensus. Extensive experiments on multiple public multi-view datasets demonstrate that our method achieves competitive downstream performance and robustness compared to prevalent TMVC methods.