A statistical learning method for simultaneous copy number estimation and subclone clustering with single-cell sequencing data.

The availability of single-cell sequencing (SCS) enables us to assess intra-tumor heterogeneity and identify cellular subclones without the confounding effect of mixed cells. Copy number aberrations (CNAs) have been commonly used to identify subclones in SCS data using various clustering methods, as cells comprising a subpopulation are found to share a genetic profile. However, currently available methods may generate spurious results (e.g., falsely identified variants) in the procedure of CNA detection, thereby diminishing the accuracy of subclone identification within a large, complex cell population. In this study, we developed a subclone clustering method based on a fused lasso model, referred to as FLCNA, which can simultaneously detect CNAs in single-cell DNA sequencing (scDNA-seq) data. Spike-in simulations were conducted to evaluate the clustering and CNA detection performance of FLCNA, benchmarking it against existing copy number estimation methods (SCOPE, HMMcopy) in combination with commonly used clustering methods. Application of FLCNA to a scDNA-seq data set of breast cancer revealed different genomic variation patterns in neoadjuvant chemotherapy-treated samples and pretreated samples. We show that FLCNA is a practical and powerful method for subclone identification and CNA detection with scDNA-seq data.

Listerin promotes cGAS protein degradation through the ESCRT pathway to negatively regulate cGAS-mediated immune response.

The enzyme cyclic GMP-AMP synthase (cGAS) is a key sensor for detecting misplaced double-stranded DNA (dsDNA) of genomic, mitochondrial, and microbial origin. It synthesizes 2'3'-cGAMP, which in turn activates the stimulator of interferon genes pathway, leading to the initiation of innate immune responses. Here, we identified Listerin as a negative regulator of cGAS-mediated innate immune response. We found that Listerin interacts with cGAS on endosomes and promotes its K63-linked ubiquitination through recruitment of the E3 ligase TRIM27. The polyubiquitinated cGAS is then recognized by the endosomal sorting complexes required for transport machinery and sorted into endosomes for degradation. Listerin deficiency enhances the innate antiviral response to herpes simplex virus 1 infection. Genetic deletion of Listerin also deteriorates the neuroinflammation and the ALS disease progress in an ALS mice model; overexpression of Listerin can robustly ameliorate disease progression in ALS mice. Thus, our work uncovers a mechanism for cGAS regulation and suggests that Listerin may be a promising therapeutic target for ALS disease.

Unveiling the role of RAC3 in the growth and invasion of cisplatin-resistant bladder cancer cells.

Bladder cancer is one of the most prevalent cancers worldwide, and its morbidity and mortality rates have been increasing over the years. However, how RAC family small GTPase 3 (RAC3) affects the proliferation, migration and invasion of cisplatin-resistant bladder cancer cells remains unclear. Bioinformatics techniques were used to investigate the expression of RAC3 in bladder cancer tissues. Influences of RAC3 in the grade, stage, distant metastasis, and survival rate of bladder cancer were also examined. Analysis of the relationship between RAC3 expression and the immune microenvironment (TIME), genomic mutations, and stemness index. In normal bladder cancer cells (T24, 5637, and BIU-87) and cisplatin-resistant bladder cancer cells (BIU-87-DDP), the expression of RAC3 was detected separately with Western blotting. Plasmid transfection was used to overexpress or silence the expression of RAC3 in bladder cancer cells resistant to cisplatin (BIU-87-DDP). By adding activators and inhibitors, the activities of the JNK/MAPK signalling pathway were altered. Cell viability, invasion, and its level of apoptosis were measured in vitro using CCK-8, transwell, and flow cytometry. The bioinformatics analyses found RAC3 levels were elevated in bladder cancer tissues and were associated with a poor prognosis in bladder cancer. RAC3 in BIU-87-DDP cells expressed a higher level than normal bladder cancer cells. RAC3 overexpression promoted BIU-87-DDP proliferation. The growth of BIU-87-DDP cells slowed after the knockdown of RAC3, and RAC3 may have had an impact on the activation of the JNK/MAPK pathway.

Radiating diversification and niche conservatism jointly shape the inverse latitudinal diversity gradient of Potentilla L. (Rosaceae).

BACKGROUND: The latitudinal diversity gradient (LDG), characterized by an increase in species richness from the poles to the equator, is one of the most pervasive biological patterns. However, inverse LDGs, in which species richness peaks in extratropical regions, are also found in some lineages and their causes remain unclear. Here, we test the roles of evolutionary time, diversification rates, and niche conservatism in explaining the inverse LDG of Potentilla (ca. 500 species). We compiled the global distributions of ~ 90% of Potentilla species, and reconstructed a robust phylogenetic framework based on whole-plastome sequences. Next, we analyzed the divergence time, ancestral area, diversification rate, and ancestral niche to investigate the macroevolutionary history of Potentilla. RESULTS: The genus originated in the Qinghai-Tibet Plateau during the late Eocene and gradually spread to other regions of the Northern Hemisphere posterior to the late Miocene. Rapid cooling after the late Pliocene promoted the radiating diversification of Potentilla. The polyploidization, as well as some cold-adaptive morphological innovations, enhanced the adaptation of Potentilla species to the cold environment. Ancestral niche reconstruction suggests that Potentilla likely originated in a relatively cool environment. The species richness peaks at approximately 45 °N, a region characterized by high diversification rates, and the environmental conditions are similar to the ancestral climate niche. Evolutionary time was not significantly correlated with species richness in the latitudinal gradient. CONCLUSIONS: Our results suggest that the elevated diversification rates in middle latitude regions and the conservatism in thermal niches jointly determined the inverse LDG in Potentilla. This study highlights the importance of integrating evolutionary and ecological approaches to explain the diversity pattern of biological groups on a global scale.

Engineering High-Tortuosity 3D Gradient Structure and CFD-Assisted Multifield Analysis for Solar Interfacial Evaporation.

Solar interfacial evaporation is a promising method for solving the global shortage of fresh water. While 2D evaporators can efficiently localize solar-converted heat at the thin layer of the water-air interface, 3D solar evaporators can maximize energy reutilization while maintaining effective mass transport ability, few studies are conducted to explore the effect of gradient porosity on evaporation performance. In this study, a multifield assisted strategy based on a gradient 3D structure with high tortuosity is proposed, which creates a thermal field environment for efficient evaporation through high absorption of sunlight and excellent photothermal conversion and uses the gradient structure to optimize the internal pressure field to enhance water evaporation and transport. This hierarchically nanostructured solar absorber, with porosity inhomogeneity-induced pressure gradient and optimized temperature management, is a valuable design idea for manufacturing a more efficient 3D solar evaporator in the field of seawater desalination. Owing to the understanding of optimizing the dimension by various simulation parameters, the evaporation efficiencies of such structures are found to be 165.7%, suppressing the most evaporator. Moreover, it can provide new ideas and references for the fields of mass transfer and thermal management.

BISC: accurate inference of transcriptional bursting kinetics from single-cell transcriptomic data.

Gene expression in mammalian cells is inherently stochastic and mRNAs are synthesized in discrete bursts. Single-cell transcriptomics provides an unprecedented opportunity to explore the transcriptome-wide kinetics of transcriptional bursting. However, current analysis methods provide limited accuracy in bursting inference due to substantial noise inherent to single-cell transcriptomic data. In this study, we developed BISC, a Bayesian method for inferring bursting parameters from single cell transcriptomic data. Based on a beta-gamma-Poisson model, BISC modeled the mean-variance dependency to achieve accurate estimation of bursting parameters from noisy data. Evaluation based on both simulation and real intron sequential RNA fluorescence in situ hybridization data showed improved accuracy and reliability of BISC over existing methods, especially for genes with low expression values. Further application of BISC found bursting frequency but not bursting size was strongly associated with gene expression regulation. Moreover, our analysis provided new mechanistic insights into the functional role of enhancer and superenhancer by modulating both bursting frequency and size. BISC also formulated a downstream framework to identify differential bursting (in frequency and size separately) genes in samples under different conditions. Applying to multiple datasets (a mouse embryonic cell and fibroblast dataset, a human immune cell dataset and a human pancreatic cell dataset), BISC identified known cell-type signature genes that were missed by differential expression analysis, providing additional insights in understanding the cell-specific stochastic gene transcription. Applying to datasets of human lung and colon cancers, BISC successfully detected tumor signature genes based on alterations in bursting kinetics, which illustrates its value in understanding disease development regarding transcriptional bursting. Collectively, BISC provides a new tool for accurately inferring bursting kinetics and detecting differential bursting genes. This study also produced new insights in the role of transcriptional bursting in regulating gene expression, cell identity and tumor progression.

Multifocal meta-fiber based on the fractional Talbot effect.

Multi-focusing of light is a crucial capability for photonic devices that can be effectively achieved by precisely modulating the phase delay on the incident wavefront. However, integrating functional structures into optical fibers for remote light focusing remains challenging due to the complex device design and limited fabrication approaches. Here, we present the design and fabrication of metalens array on the end-face of a tailored single-mode step-index fiber for focusing light field into closely packed focal spot array. The metalenses are configured based on the fractional Talbot effect and benefit a modular design capability. Light passing through the optical fiber can be focused into different focal planes. With a synergistic 3D laser nanoprinting technique based on two-photon polymerization, high-quality meta-fibers are demonstrated for focusing light parallelly with a uniform numerical aperture (NA) as high as approximately 0.77. This may facilitate various applications such as optical trapping, generation of sophisticated beam profiles, and boosting light coupling efficiencies.

Ultrahigh precision laser nanoprinting based on defect-compensated digital holography for fast-fabricating optical metalenses.

The 3D structured light field manipulated by a digital-micromirror-device (DMD)-based digital hologram has demonstrated its superiority in fast-fabricating stereo nanostructures. However, this technique intrinsically suffers from defects of light intensity in generating modulated focal spots, which prevents from achieving high-precision micro/nanodevices. In this Letter, we have demonstrated a compensation approach based on adapting spatial voxel density for fabricating optical metalenses with ultrahigh precision. The modulated focal spot experiences intensity fluctuations of up to 3% by changing the spatial position, leading to a 20% variation of the structural dimension in fabrication. By altering the voxel density to improve the uniformity of the laser cumulative exposure dosage over the fabrication region, we achieved an increased dimensional uniformity from 94.4% to 97.6% in fabricated pillars. This approach enables fast fabrication of metalenses capable of sub-diffraction focusing of 0.44λ/NA with the increased mainlobe-sidelobe ratio from 1:0.34 to 1:0.14. A 6 × 5 supercritical lens array is fabricated within 2 min, paving a way for the fast fabrication of large-scale photonic devices.

Telemedicine network latency management system in 5G telesurgery: a feasibility and effectiveness study.

BACKGROUND: Network latency is the most important factor affecting the performance of telemedicine. The aim of the study is to assess the feasibility and efficacy of a novel network latency management system in 5G telesurgery. METHODS: We conducted 20 telesurgery simulation trials (hitching rings to columns) and 15 remote adrenalectomy procedures in the 5G network environment. Telemedicine Network Latency Management System and the traditional "Ping command" method (gold standard) were used to monitor network latency during preoperative simulated telesurgery and formal telesurgery. We observed the working status of the Telemedicine Network Latency Management System and calculated the difference between the network latency data and packet loss rate detected by the two methods. In addition, due to the lower latency of the 5G network, we tested the alert function of the system using the 4G network with relatively high network latency. RESULTS: The Telemedicine Network Latency Management System showed no instability during telesurgery simulation trials and formal telesurgery. After 20 telesurgery simulation trials and 15 remote adrenalectomy procedures, the p-value for the difference between the network latency data monitored by the Telemedicine Network Latency Management System and the "Ping command" method was greater than 0.05 in each case. Meanwhile, the surgeons reported that the Telemedicine Network Latency Management System had a friendly interface and was easy to operate. Besides, when the network latency exceeded a set threshold, a rapid alarm sounded in the system. CONCLUSION: The Telemedicine Network Latency Management System was simple and easy to operate, and it was feasible and effective to use it to monitor network latency in telesurgery. The system had an intuitive and concise interface, and its alarm function increased the safety of telesurgery. The system's own multidimensional working ability and information storage capacity will be more suitable for telemedicine work.

Physically based simulation of focusing schlieren imaging for a hypersonic boundary layer flow.

Focusing schlieren systems are more advantageous than conventional schlieren systems in providing a schlieren image with certain spatial discrimination along the light path. The present work employed a hybrid of the optical-transfer matrix and ray-tracing method to faithfully replicate complete physical imaging processes throughout a focusing schlieren optic system. A direct numerical simulation of a hypersonic boundary layer flow was employed to synthesize focusing schlieren images. The influence of various configuration parameters on the properties of focusing schlieren image such as local schlieren structure, brightness, sensitivity, and depth of field were systematically explored. In addition, an approximation method was proposed as a simplified means to facilitate the simulation of a focusing schlieren image.

Exploring Sources, Biological Functions, and Potential Applications of the Ubiquitous Marine Cyclic Dipeptide: A Concise Review of Cyclic Glycine-Proline.

Cyclic glycine-proline (cGP), a prevalent marine cyclic dipeptide, possesses a distinct pyrrolidine-2,5-dione scaffold, which contributes to the chemical diversity and broad bioactivities of cGP. The diverse sources from marine-related, endogenous biological, and synthetic pathways and the in vitro and in vivo activities of cGP are reviewed. The potential applications for cGP are also explored. In particular, the pivotal roles of cGP in regulating insulin-like growth factor-1 homeostasis, enhancing neuroprotective effects, and improving neurotrophic function in central nervous system diseases are described. The potential roles of this endogenous cyclic peptide in drug development and healthcare initiatives are also highlighted. This review underscores the significance of cGP as a fundamental building block in drug discovery with exceptional drug-like properties and safety. By elucidating the considerable value of cGP, this review aims to reignite interest in cGP-related research within marine medicinal chemistry and synthetic biology.

Past climate cooling and orogenesis of the Hengduan Mountains have influenced the evolution of Impatiens sect. Impatiens (Balsaminaceae) in the Northern Hemisphere.

BACKGROUND: Impatiens sect. Impatiens is distributed across the Northern Hemisphere and has diversified considerably, particularly within the Hengduan Mountains (HDM) in southwest China. Yet, the infra-sectional phylogenetic relationships are not well resolved, largely due to limited taxon sampling and an insufficient number of molecular markers. The evolutionary history of its diversification is also poorly understood. In this study, plastome data and the most complete sampling to date were used to reconstruct a robust phylogenetic framework for this section. The phylogeny was then used to investigate its biogeographical history and diversification patterns, specifically with the aim of understanding the role played by the HDM and past climatic changes in its diversification. RESULTS: A stable phylogeny was reconstructed that strongly supported both the monophyly of the section and its division into seven major clades (Clades I-VII). Molecular dating and ancestral area reconstruction suggest that sect. Impatiens originated in the HDM and Southeast China around 11.76 Ma, after which different lineages dispersed to Northwest China, temperate Eurasia, and North America, mainly during the Pliocene and Pleistocene. An intercontinental dispersal event from East Asia to western North America may have occurred via the Bering Land Bridge or Aleutian Islands. The diversification rate was high during its early history, especially with the HDM, but gradually decreased over time both within and outside the HDM. Multiple linear regression analysis showed that the distribution pattern of species richness was strongly associated with elevation range, elevation, and mean annual temperature. Finally, ancestral niche analysis indicated that sect. Impatiens originated in a relatively cool, middle-elevation area. CONCLUSIONS: We inferred the evolutionary history of sect. Impatiens based on a solid phylogenetic framework. The HDM was the primary source or pump of its diversity in the Northern Hemisphere. Orogeny and climate change may have also shaped its diversification rates, as a steady decrease in the diversification rate coincided with the uplift of the HDM and climate cooling. These findings provide insights into the distribution pattern of sect. Impatiens and other plants in the Northern Hemisphere.

Shall genomic correlation structure be considered in copy number variants detection?

Copy number variation has been identified as a major source of genomic variation associated with disease susceptibility. With the advent of whole-exome sequencing (WES) technology, massive WES data have been generated, allowing for the identification of copy number variants (CNVs) in the protein-coding regions with direct functional interpretation. We have previously shown evidence of the genomic correlation structure in array data and developed a novel chromosomal breakpoint detection algorithm, LDcnv, which showed significantly improved detection power through integrating the correlation structure in a systematic modeling manner. However, it remains unexplored whether the genomic correlation exists in WES data and how such correlation structure integration can improve the CNV detection accuracy. In this study, we first explored the correlation structure of the WES data using the 1000 Genomes Project data. Both real raw read depth and median-normalized data showed strong evidence of the correlation structure. Motivated by this fact, we proposed a correlation-based method, CORRseq, as a novel release of the LDcnv algorithm in profiling WES data. The performance of CORRseq was evaluated in extensive simulation studies and real data analysis from the 1000 Genomes Project. CORRseq outperformed the existing methods in detecting medium and large CNVs. In conclusion, it would be more advantageous to model genomic correlation structure in detecting relatively long CNVs. This study provides great insights for methodology development of CNV detection with NGS data.

SCRIP: an accurate simulator for single-cell RNA sequencing data.

MOTIVATION: Recent advancements in single-cell RNA sequencing (scRNA-seq) have enabled time-efficient transcriptome profiling in individual cells. To optimize sequencing protocols and develop reliable analysis methods for various application scenarios, solid simulation methods for scRNA-seq data are required. However, due to the noisy nature of scRNA-seq data, currently available simulation methods cannot sufficiently capture and simulate important properties of real data, especially the biological variation. In this study, we developed scRNA-seq information producer (SCRIP), a novel simulator for scRNA-seq that is accurate and enables simulation of bursting kinetics. RESULTS: Compared to existing simulators, SCRIP showed a significantly higher accuracy of stimulating key data features, including mean-variance dependency in all experiments. SCRIP also outperformed other methods in recovering cell-cell distances. The application of SCRIP in evaluating differential expression analysis methods showed that edgeR outperformed other examined methods in differential expression analyses, and ZINB-WaVE improved the AUC at high dropout rates. Collectively, this study provides the research community with a rigorous tool for scRNA-seq data simulation. AVAILABILITY AND IMPLEMENTATION: https://CRAN.R-project.org/package=SCRIP. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Microbial autotrophy explains large-scale soil CO2 fixation.

Microbial communities play critical roles in fixing carbon from the atmosphere and fixing it in the soils. However, the large-scale variations and drivers of these microbial communities remain poorly understood. Here, we conducted a large-scale survey across China and found that soil autotrophic organisms are critical for explaining CO2 fluxes from the atmosphere to soils. In particular, we showed that large-scale variations in CO2 fixation rates are highly correlated to those in autotrophic bacteria and phototrophic protists. Paddy soils, supporting a larger proportion of obligate bacterial and protist autotrophs, display four-fold of CO2 fixation rates over upland and forest soils. Precipitation and pH, together with key ecological clusters of autotrophic microbes, also played important roles in controlling CO2 fixation. Our work provides a novel quantification on the contribution of terrestrial autotrophic microbes to soil CO2 fixation processes at a large scale, with implications for global carbon regulation under climate change.

Aberration-compensated supercritical lens for sub-diffractive focusing within 20° field of view.

The supercritical lens has shown a remarkable capability of achieving far-field sub-diffraction limited focusing through elaborating a modulated interference effect. Benefiting from the relative high energy utilization efficiency and weak sidelobe properties, the supercritical lens holds significant advantage in a series of application scenarios. However, all of the demonstrated supercritical lenses mainly work in the on-axis illumination condition, so the off-axis aberration effect will severely deteriorate its sub-diffraction limit focusing capability for the illuminating beam with an oblique angle. In this work, an aberration-compensated supercritical lens with single-layer configuration is proposed and experimentally demonstrated. Such a single-layer supercritical lens consists of multilevel phase configurations patterned with the two-photon polymerization lithography technique. The simulation and experimental recorded results show that the aberration-compensated supercritical lens with a numerical aperture value of 0.63 could achieve a far-field sub-diffraction limited focusing property within 20° field of view at a wavelength of λ = 633 nm. This monochromatic aberration-compensated supercritical lens with single-layer configuration indicates excellent potential in the development of laser scanning ultrahigh optical storage and label free super-resolution imaging.

Anthradithiophene (ADT)-Based Polymerized Non-Fullerene Acceptors for All-Polymer Solar Cells.

Realizing efficient all-polymer solar cell (APSC) acceptors typically involves increased building block synthetic complexity, hence potentially unscalable syntheses and/or prohibitive costs. Here we report the synthesis, characterization, and implementation in APSCs of three new polymer acceptors P1-P3 using a scalable donor fragment, bis(2-octyldodecyl)anthra[1,2-b : 5,6-b']dithiophene-4,10-dicarboxylate (ADT) co-polymerized with the high-efficiency acceptor units, NDI, Y6, and IDIC. All three copolymers have comparable photophysics to known polymers; however, APSCs fabricated by blending P1, P2 and P3 with donor polymers PM5 and PM6 exhibit modest power conversion efficiencies (PCEs), with the champion P2-based APSC achieving PCE=5.64 %. Detailed morphological and microstructural analysis by AFM and GIWAXS reveal a non-optimal APSC active layer morphology, which suppresses charge transport. Despite the modest efficiencies, these APSCs demonstrate the feasibility of using ADT as a scalable and inexpensive electron rich/donor building block for APSCs.

Application of Carbon Fiber-Reinforced Ceramic Composites in Active Thermal Protection of Advanced Propulsion Systems: A Review.

Thermal protection is one of the crucial issues for the advanced propulsion systems of Reusable Launch Vehicles. New service requirements for materials, such as high strength, low density, low thermal expansion, high thermal stability, etc., are raised for the thermal structure with the increasing demand of flight Mach numbers and thrust-to-weight ratio. Carbon fiber-reinforced ceramic composites, which generally meet the aforementioned requirements, show great potential for various applications and they have been widely applied in the thermal protection for hypersonic vehicles. This paper gives a comprehensive and systematic review of current research status for carbon fiber-reinforced ceramic composites applied in the thermal structure of advanced propulsion systems. Three aspects are presented and discussed: the ceramic composites fabrication and the property characterization, the thermal performance of composite thermal structure used in practical engines, and the numerical methods for predicting mechanical and thermal properties of composites as well as the physicochemical phenomenon in the cooling channels. Firstly, the main manufacturing processes for the carbon-reinforced ceramic composites are presented and the corresponding advantages and disadvantages are analyzed. The high-temperature oxidation and ablation behaviors of composites are demonstrated and the improvement of oxidation and ablation resistance by introducing the ultra-high-temperature ceramics into C/C composites is discussed in detail. Then, several typical applications of carbon fiber-reinforced ceramic composites (mainly C/SiC), including the work of RCI, JAXA and NASA, have been reviewed and analyzed. After that, the current research status of macroscale equivalent and multiscale numerical methods for predicting the mechanical and thermal properties of composites as well as the complex physicochemical phenomenon occurring in hydrocarbon fuels are sorted out. Finally, several potential prospects are pointed out for the future research on the thermal protection of advanced propulsion systems based on the carbon fiber-reinforced ceramic composites.

Model-based assessment of cardiopulmonary autonomic regulation in paced deep breathing.

Autonomic dysfunction can lead to many physical and psychological diseases. The assessment of autonomic regulation plays an important role in the prevention, diagnosis, and treatment of these diseases. A physiopathological mathematical model for cardiopulmonary autonomic regulation, namely Respiratory-Autonomic-Sinus (RSA) regulation Model, is proposed in this study. A series of differential equations are used to simulate the whole process of RSA phenomenon. Based on this model, with respiration signal and ECG signal simultaneously acquired in paced deep breathing scenario, we manage to obtain the cardiopulmonary autonomic regulation parameters (CARP), including the sensitivity of respiratory-sympathetic nerves and respiratory-parasympathetic nerves, the time delay of sympathetic, the sensitivity of norepinephrine and acetylcholine receptor, as well as cardiac remodeling factor by optimization algorithm. An experimental study has been conducted in healthy subjects, along with subjects with hypertension and coronary heart disease. CARP obtained in the experiment have shown their clinical significance.

Nacre-mimetic cerium-doped nano-hydroxyapatite/chitosan layered composite scaffolds regulate bone regeneration via OPG/RANKL signaling pathway.

Autogenous bone grafting has long been considered the gold standard for treating critical bone defects. However, its use is plagued by numerous drawbacks, such as limited supply, donor site morbidity, and restricted use for giant-sized defects. For this reason, there is an increasing need for effective bone substitutes to treat these defects. Mollusk nacre is a natural structure with outstanding mechanical property due to its notable "brick-and-mortar" architecture. Inspired by the nacre architecture, our team designed and fabricated a nacre-mimetic cerium-doped layered nano-hydroxyapatite/chitosan layered composite scaffold (CeHA/CS). Hydroxyapatite can provide a certain strength to the material like a brick. And as a polymer material, chitosan can slow down the force when the material is impacted, like an adhesive. As seen in natural nacre, the combination of these inorganic and organic components results in remarkable tensile strength and fracture toughness. Cerium ions have been demonstrated exceptional anti-osteoclastogenesis capabilities. Our scaffold featured a distinct layered HA/CS composite structure with intervals ranging from 50 to 200 µm, which provided a conducive environment for human bone marrow mesenchymal stem cell (hBMSC) adhesion and proliferation, allowing for in situ growth of newly formed bone tissue. In vitro, Western-blot and qPCR analyses showed that the CeHA/CS layered composite scaffolds significantly promoted the osteogenic process by upregulating the expressions of osteogenic-related genes such as RUNX2, OCN, and COL1, while inhibiting osteoclast differentiation, as indicated by reduced TRAP-positive osteoclasts and decreased bone resorption. In vivo, calvarial defects in rats demonstrated that the layered CeHA/CS scaffolds significantly accelerated bone regeneration at the defect site, and immunofluorescence indicated a lowered RANKL/OPG ratio. Overall, our results demonstrate that CeHA/CS scaffolds offer a promising platform for bone regeneration in critical defect management, as they promote osteogenesis and inhibit osteoclast activation.