BACKGROUND:Given the persistently high morbidity and mortality of heart failure (HF), targeting myocardial remodeling, particularly pathological hypertrophy and fibrosis, has become a major therapeutic priority. RhoA (Ras homolog gene family member A), a small GTPase governing cytoskeletal reorganization and cell migration, plays a pivotal role in this process. However, RhoA has long been considered undruggable because of its high-affinity binding to GDP/GTP and the absence of well-defined druggable pockets. METHODS:Structural analyses comparing RhoA-GTP and RhoA-GDP conformations, combined with surface plasmon resonance-based screening, were used to identify a RhoA inhibitor. The underlying mechanism was validated in cultured cells and 3-dimensional myocardial tissue models. Therapeutic efficacy was assessed across multiple species of HF models and supported by multiomics analyses linking RhoA activation to human HF. Key findings were further confirmed by multiplex immunohistochemistry and pulldown assays in human heart specimens. RESULTS:We identified an unrecognized cryptic pocket adjacent to GDP in RhoA. A natural product, AH001, selectively occupied this pocket and interacted with GDP, thereby stabilizing the interaction between RhoA and its endogenous inhibitor, RhoGDIα (Rho GDP-dissociation inhibitor 1). AH001 suppressed downstream signaling by reducing MRTFA (myocardin-related transcription factor A) nuclear translocation and downregulating fibrosis- and hypertrophy-related proteins. Moreover, AH001 disrupted pathological crosstalk between Mrtfa+ cardiomyocytes and fibroblasts. Consequently, AH001 markedly attenuated myocardial remodeling in multiple HF animal models, as well as in 3-dimensional myocardial tissue models. CONCLUSIONS:These findings establish pharmacological inhibition of RhoA activation as a viable strategy to mitigate myocardial remodeling in HF and provide a conceptual framework for developing reversible inhibitors against previously undruggable small GTPases.

The role of marginal zone B (MZB) cells in SARS-CoV-2 immunity remains unclear, particularly in comparing vaccine-induced responses to natural infection. MZB cells are crucial for rapid antibody responses, but their contributions to vaccine-induced immunity, especially in the elderly, are not fully understood. MZB-deficient (FcµR-/-) mice were immunized with SARS-CoV-2 spike protein-conjugated virus-like particles (VLPs). B cell receptor repertoire (BCR) profiles and antibody levels were assessed using ELISA, flow cytometry and single-cell sequencing. Human peripheral blood samples were collected from vaccinated individuals and unvaccinated elderly patients with Omicron infection. Vaccinated blood samples were collected at four time points: Pre-vaccine, Post-2nd dose (7 days), 6 months post-2nd, Post-booster (7 days). Infected patient samples were collected during acute and recovery phases. All samples were analyzed using hypersensitive chemiluminescence immunoassays, protein microarrays and flow cytometry. MZB cell deficiency in mice reduced splenic BCR repertoire diversity. In vaccinated individuals, total SARS-CoV-2 antibodies, including IgG, IgM and RBD-ACE2 competitive neutralization surrogate response peaked at Post-2nd dose (7 days), declined at Post-2nd dose (7 days), and increased significantly at Post-booster (7 days). Protein microarray analysis confirmed vaccine-induced antibodies targeting RBD and spike proteins. Significant expansion was observed in the percentage of MZB cells (CD21+CD23-) and follicular helper T cells (Tfh, ICOS+CXCR5+) at Post-booster (7 days), accompanied by a decrease in the percentage of follicular B cells (FoB, CD21-CD23+) in PBMCs. In contrast, unvaccinated elderly Omicron-infected individuals showed increased MZB cells during recovery compared to acute infection. MZB cells contribute to humoral immunity after both SARS-CoV-2 vaccination and natural infection. The delayed immune response in elderly individuals with natural infection underscores the importance of prioritizing vaccination for timely protection in this vulnerable group.

The rapid and discriminative detection of multiple pathogens is crucial for effective disease control, yet remains a significant challenge for point-of-care testing (POCT). Herein, we report a novel probe design strategy-Antiphasic Arctangent-Arccotangent (AAA) probes-for kinetically orthogonal, low-crosstalk multiplex detection in a single closed tube via loop-mediated isothermal amplification (LAMP). The AAA probe system comprises two kinetically complementary probes: the Arctangent Probe (ATP), a stem-loop probe that yields a rising fluorescence signal, and the Arccotangent Probe (ACP), a guanine-quenched linear probe that produces a falling fluorescence signal. Their inherently antiphasic kinetic trajectories effectively reduce false positives arising from nonspecific amplification. This system requires no additional enzymes or complex operations, preserving the simplicity of conventional LAMP. As a proof of concept, we demonstrated its utility by simultaneously detecting Japanese encephalitis virus (JEV) and Plasmodium vivax (P. vivax), two representative mosquito-borne pathogens, with high specificity and a sensitivity of 5 copies/μL. The assay performed robustly with clinical samples and in complex matrices (e.g., serum, tissue homogenates). Beyond the specific duplex detection, the AAA probe principle establishes a general framework for designing orthogonal probe sets, offering a promising and versatile strategy for multiplex nucleic acid detection in resource-limited settings.

Background Identification of Alzheimer's disease (AD) needs inexpensive, noninvasive novel biomarkers. autoimmunity plays a key role in the pathogenesis of AD. Objective This study aims to screen innovative diagnostic biomarkers for AD from the perspective of autoantibodies. Methods The study consisted of two screening phases and two validation phases. AD serum autoantibody-related biomarkers were discovered and validated using serum samples from four independent cohorts encompassing 241 participants, i.e., AD patients, healthy controls, and other dementia-related diseases. First, to identify biomarkers for AD, a phage displayed random peptide library (Ph.D.12) was applied to screen specific autoantibodies in a total of 71 serum samples from 39 AD patients and 32 healthy controls. After further screening by polypeptide microarray, Then, for validation, three peptides were analyzed in another two independent cohorts, which included AD patients, healthy controls, and other dementia-related diseases, Enzyme-linked immunosorbent assay (ELISA) was finally used to evaluate their sensitivity and specificity for AD diagnosis. Results Our results show that both AD2024Val03, AD2024Val70, and AD2024Val72 showed a statistically significant ability to discriminate AD patients from controls and other dementia-related diseases. Additionally, combination of these three peptides, AD2024Val, could greatly improve the diagnostic performance (AUC is above 0.95, sensitivity ∼100%, specificity ∼79%). Conclusions the peptides we identified could serve as promising blood biomarkers for AD clinical diagnosis, which also might provide new insights into the potential pathogenesis of AD.

KRAS is among the most frequently mutated oncogenes in human cancers, with codon 12 representing a dominant hotspot. Despite decades of study, the atomic-level conformational dynamics of KRAS and its G12 mutations remain insufficiently resolved, posing a central barrier to rational drug discovery. Elucidating these dynamics is critical for revealing hidden druggable pockets and enabling the development of mutation-specific inhibitors. Here, we delineate the impact of KRAS G12 mutations on conformational dynamics and GTP binding by performing microsecond-scale, well-tempered metadynamics simulations. This approach yielded precise free energy landscapes that expose mutation-specific alterations to structural states. By introducing biologically meaningful collective variables, including key angles (ϕ) and distances (d1, d2) defining Switch-I, Switch-II, and the P-loop, we identified distinct local and global minima as well as their transition states. Analysis of these transitions reveals how G12 mutations reshape GTP affinity and drive aberrant KRAS activation. We further demonstrated the convergence of the simulations and discussed the physiological relevance of the calculation results in comparison to previously published experimental and theoretical data. Importantly, our analyses uncovered druggable pockets unique to nonpolar G12 mutants, offering novel entry points for inhibitor design. Collectively, this work provides an atomic-level framework for understanding KRAS G12 mutations, establishes transferable collective variables for broader RAS family studies, and opens new avenues for mutation-specific therapeutic development.

Abstract Mimotopes selected by phage display have been used for antibody epitope mapping for nearly four decades, but residue-level recovery of functional epitopes has remained unreliable. This is not a problem of algorithms but of assumptions: mimotopes recapitulate native epitope recognition through compensatory interaction networks that preserve binding energetics, without conserving sequence or surface geometry. Predictors trained on sequence similarity or static interface geometry therefore miss the residues that drive recognition. We present a physics-based workflow that refines antibody-bound mimotope conformations by microsecond-scale molecular dynamics and maps the resulting structures onto the cognate antigen using Folddisco. Across four antibody–antigen systems with crystallographically defined epitopes, the workflow reaches 60 to 100% precision in epitope localization and recovers 100% of mutagenesis- or structurally validated functional hotspots, compared with at most 25% for three widely used benchmarks (EpiSearch, ClusPro, SEPPA-mAb). For the clinically relevant ChiLob 7/4 system, it further identifies a minimal tetrapeptide (PWVP) sufficient for antibody recognition, validated by Western blot and immunoprecipitation. By grounding epitope prediction in experimentally selected binding information and energy-resolved conformational refinement, the workflow connects phage display to mechanistic structural insight.

Xiamenmycin, a prenylated benzopyran compound produced by Streptomyces xiamenensis 318, exhibits promising anti-fibrotic activity but suffers from low productivity, limiting its pharmaceutical development. In this study, we obtained a mutant strain N-8 producing 226.3 mg/L xiamenmycin B (28-fold increase relative to the parent strain MT-XN) via N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. Further iterative ARTP treatments yielded mutant strain A5-8 with a titer of 429.4 mg/L. This screening was achieved using a fused antibiotic marker (neo) downstream of the xiamenmycin biosynthetic genes, which enabled kanamycin resistance-based selection. Furthermore, intracellular precursors of xiamenmycin were increased, as indicated by transcriptome analysis to be due to upregulation of biosynthetic pathway genes. Genome re-sequencing revealed deletions of ximA and a 138 bp upstream region of ximB promoter in strain N-8. In situ complementation of the 138 bp region significantly reduced xiamenmycin B production. Mechanistically, we identified a TetR-family transcription factor, XimN1, that specifically binds to the upstream of the ximB promoter and exerts a negative regulatory effect. Deletion of this region alleviated XimN1-mediated repression, upregulates ximBCDE transcription, and enhances xiamenmycin B production. The upstream region also contains the binding sites for AdpA. EMSA analysis further revealed that both proteins bind within the 138 bp region. In summary, we successfully constructed high-yield xiamenmycin mutants and elucidated a novel trans-acting transcription factor, XimN1 control xiamenmycin biosynthesis. This study presents practical strategies for improving the production efficiency of metabolites and offers novel insights into the transcriptional regulatory underlying secondary metabolism in Streptomyces.

African swine fever virus (ASFV) has a large, highly variable genome; single-gene assays therefore risk false-negative results, and a field-deployable, multi-target nucleic acid test is urgently needed. Here we present DeepLAMP, a platform that combines a hive-shaped microfluidic chip, gold-nanoparticle-enhanced colorimetric loop-mediated isothermal amplification (LAMP), and smartphone-based deep-learning analysis to synchronously detect four conserved ASFV genes (B646L, Q706L, P1192R and B475L). The AuNP-catalyzed micro-environment markedly boosts amplification efficiency, lowering limits of detection to 5 copies mu L-1 for B646L and P1192R, and to 25 and 50 copies mu L-1 for Q706L and B475L, respectively; the full assay is completed within 60 min. An optimized ConvNeXt deep learning model classifies six chip image categories with 98.6 % overall accuracy, automatically quantifying color shifts imperceptible to the naked eye. DeepLAMP correctly identified clinical ASFV samples and showed no cross reactivity with CSFV, PRRSV or PRV, demonstrating high specificity and robustness. Requiring neither centrifugation nor costly instrumentation, the workflow delivers "sample-to-answer" operation in pig farms and other resource-limited settings, offering a versatile point-of-care platform readily extendable to multi-gene diagnostics of other zoonotic and emerging pathogens.

Antibodies play crucial roles in both basic research and clinical practice. The binding interface between an antibody and its antigen, i.e., the epitope, is a defining feature of the antibody. Accurate epitope mapping is essential for developing antibody-based reagents and therapies and safeguarding intellectual property. Here, we present the detailed procedure of a novel high-throughput technology, namely, Antibody binding epitope Mapping (AbMap) version 1.0. Leveraging versatile phage-displayed peptide library and next-generation sequencing (NGS), AbMap enables the comprehensive profiling of hundreds to thousands of antibody-binding epitopes in a single run.

SARS-CoV-2 infection and vaccination both trigger immune responses. The former leads to naturally acquired immunity, while the latter induces active immunity through artificial means. However, the distinct immune effects of vaccination and infection, as well as their underlying mechanisms, require further clarification. In this study, we compared the peripheral B cell differentiation, serological differences and the expression level of BCR signaling molecules between the vaccinated and recovered group. The vaccinated group exhibited reduced RBD-specific B cell differentiation and lower CD86 signal intensity on memory B cells, but enhanced BCR signaling in B cells. Regarding metabolic signaling, the vaccinated group had elevated expression levels of pS6, c-Myc, pmTOR, and pSTAT5, suggesting that the STAT5-c-Myc axis plays a role in regulating B cell metabolism. Additionally, proteome microarray analysis revealed that the serum of the vaccinated group contained higher levels of IgG antibodies against the SARS-CoV-2 N-Nter protein and IgA antibodies specific to the SARS-CoV-2 S1 protein. In summary, these findings indicate that the vaccinated group develops a more robust coronavirus-specific immune response, with enhanced BCR signaling and metabolic activity compared to the recovered group. These insights might contribute to the optimization of SARS-CoV-2 vaccine design.

Developing an efficient and safe vaccine for African swine fever (ASF), a devastating disease of pigs, remains a significant challenge mainly due to limited knowledge of the immune correlates of protection. Identifying protective determinants is difficult because ASF virus (ASFV) is a large and complex DNA virus encoding over 160 proteins. Here, we constructed an ASFV proteome microarray containing 160 full-length proteins for profiling ASFV-specific antibodies. An antibody reactome containing 46 ASFV proteins (including 12 newly recognized B-cell antigens) was established by analyzing several cohorts of serum samples from pigs protected with different live-attenuated vaccines (LAVs). A proteome-wide study of antibody dynamics over a 26-day period provided a multi-dimensional landscape of the host humoral response against ASFV after acute infection, LAV immunization, and post-vaccination challenge. This study provides a comprehensive understanding of ASFV-induced humoral immune responses, highlights B-cell antigen candidates for vaccine design, supports the investigation of LAV protection mechanisms, and would accelerate vaccine development.IMPORTANCEAfrican swine fever (ASF) poses a severe threat to global swine industries, with vaccine development hindered by limited understanding of immune protection. A comprehensive understanding of antibody responses against ASF virus (ASFV) and the discovery of protective antigens are fundamental to vaccine development. This study constructed an ASFV proteome microarray to profile antibody responses against 160 viral proteins and established the antibody spectra against ASFV with dynamic features. The proteome microarray offers a high-throughput platform for understanding ASFV immunology and pathogenicity and will contribute to ASF vaccine development and diagnosis.

Lipid metabolism reprogramming is critical for the initiation and progression of hepatocellular carcinoma (HCC). However, how the dysregulation of lipid metabolism contributes to HCC development remains largely unknown. Here, we report that the m6A reader YTHDC1-mediated epigenetic regulation of the long noncoding RNA NEAT1 activates stearoyl-CoA desaturase (SCD)-associated lipid metabolic processes during HCC progression. Mechanistically, histone lactylation in HCC induces increased expression of YTHDC1, increasing the stability of m6A-modified NEAT1. The histone acetyltransferase p300 is then recruited by NEAT1 and activates SCD by increasing the level of histone acetylation at the SCD promoter, thus facilitating HCC progression via hepatocellular lipid metabolism remodeling. Taken together, these discoveries suggest a close link between the epigenetic machinery and lipid metabolic abnormalities, which promotes cancer progression.

Allergic asthma is a chronic airway disease characterized by an allergic response and altered immune tolerance. CD4+ tissue-resident memory T (TRM) cells are crucial in the chronic and relapsing pathogenesis of asthma. Furthermore, promyelocytic leukemia zinc finger (PLZF) is an essential transcription factor involved in asthmatic tolerance and has been implicated in the regulation of CD4+CD44+ memory T cells. However, the role of CD4+ TRM cells in asthmatic tolerance, as well as their potential modulation by PLZF, remain unclear. Therefore, in the current study, we explore the role of CD4+ TRM cells in asthmatic immune tolerance and as well as the regulatory role of PLZF in this process. To elucidate the role of CD4+ TRM cells in immune tolerance, asthma memory mouse models were treated with the immunomodulator FTY720. Subsequently, CD4+ T cells were isolated from the lungs and spleens and transferred to oral tolerance mouse models. To explore the regulation of PLZF in CD4+ TRM cells, asthma and oral tolerance were established in Zbtb16flox/flox CD4Cre and wild-type mice. Flow cytometry, histological analysis, and cytokine measurements were performed to characterize the immune response. The regulatory activity of PLZF on CD4+ TRM cells was analyzed through quantitative proteomics and verified in vitro and vivo. The CD4+ TRM cell proportion positively correlated with the pathological phenotypes and molecular characteristics of asthma. Adoptive transfer of CD4+ TRM cells induced asthmatic phenotypes. This suggested that CD4+ TRM cells contributed to the pathogenesis of asthma. Conditional knockout of PLZF substantially reduced the proportion of CD4+ TRM cells, relieved asthmatic symptoms, and suppressed the interleukin (IL)-15/IL-15Rα signaling pathway. Furthermore, exposure to the IL-15Rα agonist restored asthma-related Th2 inflammation, accompanied by a markedly increased proportion of CD4+ TRM cells. Meanwhile, IL-15 and ovalbumin(OVA)-primed Beas2b supernatant co-stimulation in vitro enhanced the differentiation of pulmonary PLZF-expressing CD4+ T cells into CD4+ TRM cells. This study identified CD4+ TRM cells as key mediators of immune tolerance in asthma. This process is regulated by the transcription factor PLZF in CD4+ T cells through IL-15/IL-15Rα signaling. Thus, targeting PLZF or the IL-15/IL-15Rα pathway may represent a promising therapeutic strategy for treating asthma.

Cyclic diguanosine monophosphate (c-di-GMP) is a ubiquitous bacterial secondary messenger with diverse functions. A previous Escherichia coli proteome microarray identified that c-di-GMP binds to the 23S rRNA methyltransferases RlmI and RlmE. Here we show that c-di-GMP inhibits RlmI activity in rRNA methylation assays, and that it modulates ribosome assembly in the presence of kanamycin. Molecular dynamics simulation and mutagenesis studies reveal that c-di-GMP binds to RlmI at residues R64, R103, G114, and K201. Structural simulations indicate that c-di-GMP quenches RlmI activity by inducing the closure of the catalytic pocket. We also show that c-di-GMP promotes antibiotic tolerance through RlmI. Binding and methylation assays indicate that the inhibitory effect of c-di-GMP on RlmI is conserved across various pathogenic bacteria. Our data suggest an unexpected role for c-di-GMP in regulating ribosome assembly under stress through the inhibition of rRNA methyltransferases.

While affinity purification-mass spectrometry (AP-MS) has significantly advanced protein-protein interaction (PPI) studies, its limitations in detecting weak, transient, and membrane-associated interactions remain. To address these challenges, we introduced a proteomic method termed affinity purification coupled proximity labeling-mass spectrometry (APPLE-MS), which combines the high specificity of Twin-Strep tag enrichment with PafA-mediated proximity labeling. This method achieves improved sensitivity while maintaining high specificity (4.07-fold over AP-MS). APPLE-MS also revealed the dynamic mitochondrial interactome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ORF9B during antiviral responses, while endogenous PIN1 profiling uncovered novel roles in DNA replication. Notably, APPLE-MS enabled in situ mapping of GLP-1 receptor complexes, demonstrating its unique capabilities for membrane PPI studies. This versatile method advances interactome research by providing comprehensive, physiologically relevant PPI networks, opening new opportunities for mechanistic discovery and therapeutic targeting.

High rates of heart failure (HF) morbidity and mortality have made targeting myocardial remodeling—particularly hypertrophy and fibrosis—a key therapeutic focus. RhoA, which regulates cytoskeletal reorganization and cell migration, plays a role in this process. However, RhoA has long been considered “undruggable”, due to its strong binding to its endogenous substrates, GDP/GTP, and the lack of well-defined pockets for drug targeting. Here, we discovered a cryptic pocket proximate to GDP within RhoA and identified a natural product, AH001, binds here and interacts with GDP, stabilizing RhoA’s interaction with its endogenous inhibitor, RhoGDIα. AH001 reduced the downstream MRTFA nuclear translocation and downregulated fibrosis/hypertrophy proteins. Consequently, AH001 mitigated myocardial remodeling in multiple HF animal models, and in the 3D myocardial tissue model. Our findings highlight the therapeutic potential of inhibiting RhoA activation in myocardial remodeling, ultimately targeting HF, and offer a promising avenue for developing reversible inhibitors against undruggable GTPases. ### Competing Interest Statement The authors have declared no competing interest. the National Natural Science Foundation of China, 82425104, 82222075 the National Key Research and Development Program of China, 2022YFC3400501 the Science and Technology Commission of Shanghai Municipality, 24JS2830200 the Shanghai Municipal Education Commission, 2024AI01014

African swine fever virus (ASFV) is a large and structurally complex DNA virus encoding more than 160 proteins, including more than 68 structural proteins. A protein library covering recombinant ASFV proteins is fundamentally important for studies on protein function, antigenicity, vaccine development, and virus-host interactions. Here, to construct an ASFV protein library, we add a glutathione S-transferase (GST) tag at the N-terminus of each ASFV protein to facilitate solubilization and purification and express the recombinant proteins in the yeast host. By optimizing codons, expression vectors and strains and conditions of expression and purification, we achieve satisfactory protein yields for analytical applications and maximized access to the whole proteome of ASFV, with coverage of ca. 95%. Using the library, a protein chip is constructed and used to screen for interactions between ASFV and swine proteins ( e.g., IRF3, p65, and IκBα). The ASFV protein library lays the groundwork for understanding and combatting ASFV. The methods for constructing the library are instructive for generating other protein libraries for high-throughput applications.

While affinity purification-mass spectrometry (AP-MS) has significantly advanced protein-protein interaction (PPI) studies, its limitations in detecting weak, transient, and membrane-associated interactions remain. To address these challenges, we introduced an innovative proteomic method termed Affinity Purification coupled Proximity LabEling-Mass Spectrometry (APPLE-MS), which combines the high specificity of Twin-Strep-tag enrichment with PafA-mediated proximity labeling. This method achieves unprecedented sensitivity while maintaining high specificity (4.07-fold over AP-MS). APPLE-MS also revealed the dynamic mitochondrial interactome of SARS-CoV-2 ORF9B during antiviral responses, while endogenous PIN1 profiling uncovered novel roles in DNA replication. Notably, APPLE-MS enabled in situ mapping of GLP-1 receptor complexes, demonstrating its unique capabilities for membrane PPI studies. This versatile method advances interactome research by providing comprehensive, physiologically relevant PPI networks, opening new opportunities for mechanistic discovery and therapeutic targeting. ![Figure][1] MOTIVATION – Cell Reports Methods only Affinity purification-mass spectrometry (AP-MS) has become a widely used method for capturing high-affinity protein-protein interactions (PPIs). However, it still faces several limitations, including high levels of non-specific binding, challenges in detecting weak interactions, and limited ability to identify PPIs at the cell surface in situ. To overcome these challenges, we introduce a modular AP-MS method enhanced by PafA-mediated proximity labeling, which improves both the specificity and sensitivity of PPI detection in a single, streamlined workflow. SIGNIFICANCE – Cell Chemical Biology only Proximity labeling is a powerful tool in chemical biology, providing critical support for studying protein function, cell signaling, and disease mechanisms by chemically labeling and capturing biomolecular interactions. In this study, we introduce a modular affinity purification-mass spectrometry (AP-MS) method that integrates proximity labeling. This method leverages the proximity-dependent enzymatic activity of PafA to covalently attach PupE to nearby proteins, in combination with the high-affinity interaction between Twin-Strep-tag and streptavidin to achieve efficient binding. This innovative approach enables precise identification of protein-protein interactions (PPIs), offering a powerful tool for capturing weak and transient interactions that are often missed by traditional AP-MS techniques. By enhancing both the specificity and efficiency of protein labeling, our work provides a robust chemical biology strategy to elucidate complex protein interactions, which are essential for understanding cellular processes and developing targeted therapeutic strategies, offering new insights into protein function and interaction dynamics in both health and disease. ### Competing Interest Statement The authors have declared no competing interest. Lingang Laboratory (Startup Fund) the Fourteenth Five-Year National Key Research and Development Program of China, 2023YFC2307200 the Natural Science Foundation of China, 92374110, 32271492 the R&D Program of Guangzhou National Laboratory, GZNL2023A01005 [1]: pending:yes