Surface Topography Modulates Cell Mechanosensing of Extracellular Matrix

The interaction between the cell membrane and the extracellular matrix is crucial for many cellular functions by modulating mechanosensitive signaling pathways.Physical properties of the extracellular matrix such as stiffness and topography affect such interactions.Our recent work reveals that surface topography of tens to hundreds of nanometer scale modulates cell signaling by activating intracellular curvature-sensitive proteins.We use vertical nanostructures protruding from a flat surface as a platform to induce precise curvatures on the cell membrane and to probe biological processes in live cells.Vertical nanopillars deform the plasma membrane inwards and induce membrane curvature when the cell engulfs them,leading to a reduction of the membrane-substrate gap distance.We found that the high membrane curvature induced by vertical nanopillars significantly affects the distribution of curvature-sensitive proteins and stimulates several cellular processes in live cells including cellular endocytosis and cytoskeleton dynamics.Our studies show a strong interplay between biological cells and nano-featured surfaces,which is an essential consideration for future development of interfacing devices.

CD98 modulates alloimmune responses:evidence from the CD98-deficient mouse

CD98 heavy chain（CD98hc）,encoded by Slc3a2,is a widely expressed vertebrate membrane protein whose functions are known as facilitating amino acid transporter and mediating integrin signaling.Little is known about its function on T lymphocyte mediated immune response to alloantigen. Here we report that we successfully deleted CD98hc in T cells by crossing mice bearing a loxP-flanked Slc3a2 allele with those expressing Cre re-combinase in T cells（CD4-Cre+）.T cell-specific deficient of CD98hc resulted in lower responses to alloantigen stimulation in mixed lymphocyte reaction assay.Heterotopic cardiac grafting was then performed from BALB/c（H-2Kd） to CD98hclox/-CD4-Cre+ /C57BL/6（H-2Kb） or control littermate C57BL/6 （B6） mice.We found that all CD98hclox/-CD4-Cre+ recipients had indefinite survival（MST:】100days, n=8）.In contrast,all littermate B6 recipients suffered acute rejection[MST:（7.4±0.5）d,n=12].In addition,the survival of the skin grafts from donor BALB/c mice to more than postoperative day （POD） 100 heart-bearing tolerant CD98hclox/-CD4- Cre+ recipients was significantly prolonged[MST: （15.2±2.2）d,n =5]compared that of the B6 mice [MST:（8.2±1.3）d,n=9].In consistent with graft survival, we found that the graft infiltration cells on POD7 were fewer than that of the B6 mice by FACS and immune-staining analysis.Also chemotaxis assay data revealed that the migrated CD98hclox/-CD4-Cre+ lymphocyte were less than that of B6 in the presence of different concentration of chemokines CCL2, CCL5,and CCL2 plus CCL5.In addition,a neutralizing antibody（clone 26-24）specific against CD98hc prolonged the graft survival[MST:（13.4±2.7）d,n=8; P=0.001]in the B6 recipients after they received the BALB/c mice heart.Hence our data indicated that T cell-specific deficient of CD98hc impaired proliferate in response to alloantigens and decreased migration ability result in inducing immune tolerance after cardiac transplantation.Moreover,the application of the blocked of CD98hc by monoclonal antibody is effective in the treatment of acute cardiac allograft rejection, which may be useful clinically in the future.

Nimodipine Modulates Bcl-2 and Bax mRNA Expression after Cerebral Ischemia

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA. The increased ratio of Bcl-2 and Bax mRNA may contribute to the anti-apoptic effect of nimodipine. The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new strategy to manage neuronal damage.

Liposome cocktail activator modulates hepatocytes and remodels the microenvironment to mitigate acute liver failure

Acute liver failure(ALF)is a mortal and critical hepatic disease,in which oxidative stress,inflammation storm and hepatocyte death are crucial in the pathogenesis.Hence,in contrast to the control of a single link,a combination therapy targeting multiple pathogenic links of the disease will be a favorable means to control the progression of the disease.In this study,we constructed dimethyl itaconate-loaded liposomes modified with dodecyl gallate as a cocktail activator to investigate its functional role in acetaminophen(APAP)-induced ALF.Our results demonstrated that the cocktail activator acted on hepatocytes and triggered cocktail efficacy,thereby simultaneously attenuating APAP-induced hepatocyte damage and remodeling the damage microenvironment.The cocktail activator could effectively scavenge reactive oxygen species,inhibit excessive inflammatory responses and reduce cell death in impaired hepatocytes for detoxification.More importantly,the cocktail activator could remodel the damage microenvironment,thus further promoting hepatocyte expansion and specifically switching macrophages from the M1 to M2 phenotype for a favorable liver regeneration of ALF.Furthermore,in APAP-induced ALF mouse model,the cocktail activator improved liver function,alleviated histopathological damage and increased survival rate.In summary,these findings indicate that the cocktail activator may provide a promising therapeutic approach for ALF treatment as a nanomedicine.

Therapeutic Administration of <i>Mycobacterium bovis</i>BCG Killed by Extended Freeze-Drying Modulates Airway Inflammation in a Chronic Murine Model of Asthma

Background: We previously showed that treatment with Mycobacterium bovis BCG killed by extended freeze-drying (EFD BCG) modulates inflammation through regulatory T cells (Tregs) in an acute asthma model. In this study, we investigated the kinetics of Treg induction as well as their long-term homing in spleen and lungs correlating with reduced airway hyperresponsiveness (AHR) in a murine model of acute allergic asthma. We then evaluated the therapeutic implication of EFD BCG in a chronic asthma model. Methods: Tregs expressing Foxp3 were analyzed in various organs shortly and long-term after EFD BCG, live- and Heat Killed-(HK-) BCG treatments in an acute model of asthma. We further studied EFD BCG treatment on airway inflammation using a chronic model of asthma in mice. Results: Foxp3 expression peaked in the inguinal draining lymph-nodes (iDLNs) 2-4 days after EFD BCG treatment whereas it was long-term observed in spleen (days 7 to 90). This increase in Foxp3 expression was also found in lungs upon intranasal ovalbumin (OVA) challenge in OVA-sensitized mice. The loss of protection 4 months after EFD BCG treatment was correlated with the end of this phenomenon. Moreover, major lung inflammation hallmarks of severe asthma after multiple allergen challenges promoting chronic airway inflammation in OVA sensitized mice were reduced by EFD BCG treatment: AHR, eosinophils and neutrophils in bronchoalveolar lavage (BAL), mucus metaplasia, Th2 as well as Th17 cytokine levels in BAL and sera. EFD BCG treatment also enhances PPAR-γ expression and regulates NF-κBp65 translocation in lung extracts in this model of chronic asthma. Conclusions: EFD BCG treatment induced long-term protective effect associated to Foxp3 Tregs in the spleen and lungs in an acute model of asthma and inhibits AHR in a chronic model of asthma. EFD BCG could be a new and promising immuno-modulatory alternative treatment to corticoids in severe human asthma.

Reciprocal inhibition of expression between RAV1 and BES1 modulates plant growth and development in Arabidopsis

RAV1(Related to ABI3/VP1)is a plant-specific B3 and AP2 domain-containing transcription factor that acts as a negative regulator of growth in many plant species.The expression of RAV1 is downregulated by brassinosteroids(BRs);large-scale transcriptome analyses have shown that the expression of RAV1 was previously targeted by BRI1-EMS-SUPPRESOR1(BES1)and BRASSINAZOLE-RESISTANT1(BZR1),which are critical transcription factors for the BR-signaling process.Using RAV1-overexpressing transgenic plants,we showed that RAV1 overexpression reduced the BR signaling capacity,resulting in the downregulation of BR biosynthetic genes and BES1 expression.Furthermore,we demonstrated that BES1,not BZR1,is directly bound to the RAV1 promoter and repressed RAV1 expression,and vice versa;RAV1 is also bound to the BES1 promoter and repressed BES1 expression.This mutual inhibition was specific to RAV1 and BES1 because RAV1 exhibited binding activity to the BZR1 promoter but did not repress BZR1 expression.We observed that constitutively activated BR signaling phenotypes in bes1-D were attenuated by the repression of endogenous BES1 expression in transgenic bes1-D plants overexpressing RAV1.RNA-sequencing analysis of RAV1-overexpressing transgenic plants and bes1-D mutant plants revealed differentially expressed genes by RAV1 and BES1 and genes that were oppositely co-regulated by RAV1 and BES1.RAV1 and BES1 regulated different transcriptomes but co-regulated a specific set of genes responsible for the balance between growth and defense.These results suggested that the mutual inhibitory transcriptional activities of RAV1 and BES1 provide fine regulatory mechanisms for plant growth and development.

A Novel Multiple DBC-staked units Package to Parallel More Chips for SiC Power Module

Silicon carbide(SiC) power modules play an essential role in the electric vehicle drive system. To improve their performance, reduce their size, and increase production efficiency, this paper proposes a multiple staked direct bonded copper(DBC) unit based power module packaging method to parallel more chips. This method utilizes mutual inductance cancellation effect to reduce parasitic inductance. Because the conduction area in the new package is doubled, the overall area of power module can be reduced. Entire power module is divided into smaller units to enhance manufacture yield, and improve design freedom. This paper provides a detailed design, analysis and fabrication procedure for the proposed package structure. Additionally, this paper offers several feasible solutions for the connection between power terminals and DBC untis. With the structure, 18dies were paralleled for each phase-leg in a econodual size power module. Both simulation and double pulse test results demonstrate that, compared to conventional layouts, the proposed package method has 74.8% smaller parasitic inductance and 34.9% lower footprint.

基于SGP4模型的空间站轨道精度分析

针对在地面站天线对空间站观测任务中,通常基于卫星工具包(satellite tool kit,STK)软件规划出天线对空间站的方位角和俯仰角,实现天线对空间站的自动跟踪.为了保证天线跟踪的准确性和可靠性,需要定期计算出准确的空间站轨道和天线的方位俯仰角,并更新规划任务.因此科学分析与评估空间站两行轨道根数(two line elements,TLE)长期预报精度,对地面站实现空间站的精准跟踪具有重要意义.本文以中国空间站(China Space Station,CSS)梦天实验舱为例,基于TLE数据,利用STK软件提供的简化常规摄动规模型(simplified general perturbations4,SGP4)模型计算空间站轨道以及空间站相对于西安地面站的方位角和俯仰角,并分析不同策略下的精度效果.试验结果表明:在第二天更新空间站的TLE,可以获得较好的轨道结果,从而更好地保障天线的跟踪精度.

THESEUS1 positively modulates plant defense responses against Botrytis cinerea through GUANINE EXCHANGE FACTOR4 signaling

The plant cell wall is an important interface for sensing pathogen attack and activating signaling pathways that promote plant immune responses.THESEUS1(THE1) acts as a sensor of cell wall integrity that controls cell elongation during plant growth.However, no specific role for THE1 in plant defense responses has been reported. Here, we found that THE1 interacts with GUANINE EXCHANGE FACTOR4(GEF4)and that both proteins play regulatory roles in plant resistance to the necrotrophic fungus Botrytis cinerea.Genetic analysis showed that THE1 and GEF4 function in the same genetic pathway to mediate plant defense responses. In addition, using transcriptome analysis, we identified various genes(such as defense-related,secondary metabolite-related, and transcription factor genes) that are likely downstream targets in the THE1-GEF4 signaling pathway. Our results suggest that THE1 functions as an upstream regulator of GEF4 signaling to positively regulate defense responses against B. cinerea in Arabidopsis.

面向石墨电极标签识别的轻量化网络LGSNet

轻量化网络已成为面向工业场景部署的关键技术。为进一步提升Ghost module的特征提取能力并减少参数量,提出了一种改进的S-Ghost瓶颈模块(Small Ghost Bottleneck)。此瓶颈模块采用1×1卷积通道与Ghost module并联的结构,缩减Ghost module的通道数以压缩参数量,并用与之并联的1×1卷积进行通道扩充;在模块输出端引入通道混洗(Channel Shuffle)操作以保证通道间信息交互。实验结果表明,利用该瓶颈结构设计的图像分类网络LGSNet (Light Ghost Networks,LGSNet),其计算量和参数量显著降低,同时网络精度与推理速度未受影响,甚至在一些测试中取得最优,此网络设计满足工业需求。这为面向工业场景的轻量化网络架构设计提供了新的解决方案和思路。

Influence of substrate effect on near-field radiative modulator based on biaxial hyperbolic materials

Relative rotation between the emitter and receiver could effectively modulate the near-field radiative heat transfer(NFRHT)in anisotropic media.Due to the strong in-plane anisotropy,natural hyperbolic materials can be used to construct near-field radiative modulators with excellent modulation effects.However,in practical applications,natural hyperbolic materials need to be deposited on the substrate,and the influence of substrate on modulation effect has not been studied yet.In this work,we investigate the influence of substrate effect on near-field radiative modulator based onα-MoO_(3).The results show that compared to the situation without a substrate,the presence of both lossless and lossy substrate will reduce the modulation contrast(MC)for different film thicknesses.When the real or imaginary component of the substrate permittivity increases,the mismatch of hyperbolic phonon polaritons(HPPs)weakens,resulting in a reduction in MC.By reducing the real and imaginary components of substrate permittivity,the MC can be significantly improved,reaching 4.64 forε_(s)=3 at t=10 nm.This work indicates that choosing a substrate with a smaller permittivity helps to achieve a better modulation effect,and provides guidance for the application of natural hyperbolic materials in the near-field radiative modulator.

Design of Protograph LDPC-Coded MIMO-VLC Systems with Generalized Spatial Modulation

This paper investigates the bit-interleaved coded generalized spatial modulation(BICGSM) with iterative decoding(BICGSM-ID) for multiple-input multiple-output(MIMO) visible light communications(VLC). In the BICGSM-ID scheme, the information bits conveyed by the signal-domain(SiD) symbols and the spatial-domain(SpD) light emitting diode(LED)-index patterns are coded by a protograph low-density parity-check(P-LDPC) code. Specifically, we propose a signal-domain symbol expanding and re-allocating(SSER) method for constructing a type of novel generalized spatial modulation(GSM) constellations, referred to as SSERGSM constellations, so as to boost the performance of the BICGSM-ID MIMO-VLC systems.Moreover, by applying a modified PEXIT(MPEXIT) algorithm, we further design a family of rate-compatible P-LDPC codes, referred to as enhanced accumulate-repeat-accumulate(EARA) codes,which possess both excellent decoding thresholds and linear-minimum-distance-growth property. Both analysis and simulation results illustrate that the proposed SSERGSM constellations and P-LDPC codes can remarkably improve the convergence and decoding performance of MIMO-VLC systems. Therefore, the proposed P-LDPC-coded SSERGSM-mapped BICGSMID configuration is envisioned as a promising transmission solution to satisfy the high-throughput requirement of MIMO-VLC applications.

High photoelectric conversion efficiency and fast relaxation time of FA_(0.4)MA_(0.6)PbI_(3) applied in ultrafast modulation of terahertz waves

Active control of terahertz(THz)waves is attracting tremendous attentions in terahertz communications and active photonic devices.Perovskite,due to its excellent photoelectric conversion performance and simple manufacturing process,has emerged as a promising candidate for optoelectronic applications.However,the exploration of perovskites in optically controlled THz modulators is still limited.In this work,the photoelectric properties and carrier dynamics of FA_(0.4)MA_(0.6)PbI_(3)perovskite films were investigated by optical pumped terahertz probe(OPTP)system.The ultrafast carrier dynamics reveal that FA_(0.4)MA_(0.6)PbI_(3)thin film exhibits rapid switching and relaxation time within picosecond level,suggesting that FA_(0.4)MA_(0.6)PbI_(3)is an ideal candidate for active THz devices with ultrafast response.Furthermore,as a proof of concept,a FA_(0.4)MA_(0.6)PbI_(3)-based metadevice with integrating plasma-induced transparency(PIT)effect was fabricated to achieve ultrafast modulation of THz wave.The experimental results demonstrated that the switching time of FA_(0.4)MA_(0.6)PbI_(3)-based THz modulator is near to 3.5 ps,and the threshold of optical pump is as low as 12.7μJ cm^(-2).The simulation results attribute the mechanism of ultrafast THz modulation to photo-induced free carriers in the FA_(0.4)MA_(0.6)PbI_(3)layer,which progressively shorten the capacitive gap of PIT resonator.This study not only illuminates the potential of FA_(0.4)MA_(0.6)PbI_(3)in THz modulation,but also contributes to the field of ultrafast photonic devices.

Physical exercise and traumatic brain injury: is it question of time?

Is it better to be safe than sorry?This Hamletic dilemma has always stimulated medical-scientific debates in numerous fields of biomedicine.And among these,the preventive-therapeutic approach to the treatment of brain trauma is one of the most striking examples.Traumatic brain injury(TBI)is a leading cause of brain damage among young and elderly populations with a very high hospitalization and death rate.TBI is characterized by two pathologically distinct but strictly consequential phases:a first characterized by an immediate and highly variable mechanical dysfunction of the brain tissue,which involves widespread cell death and tissue degeneration,followed by a second phase which can last from days to even years depending on the severity of the TBI and the patient’s pre-existing health status.Secondary processes,including inflammatory phenomena,oxidative stress associated with metabolic,vascular,and neuro-modulatory deficits,are very often responsible for neuro-motor and psychological deficits leading to long-term disabilities(Kaur and Sharma,2018).

Nonreciprocal thermal metamaterials:Methods and applications

Nonreciprocity of thermal metamaterials has significant application prospects in isolation protection,unidirectional transmission,and energy harvesting.However,due to the inherent isotropic diffusion law of heat flow,it is extremely difficult to achieve nonreciprocity of heat transfer.This review presents the recent developments in thermal nonreciprocity and explores the fundamental theories,which underpin the design of nonreciprocal thermal metamaterials,i.e.,the Onsager reciprocity theorem.Next,three methods for achieving nonreciprocal metamaterials in the thermal field are elucidated,namely,nonlinearity,spatiotemporal modulation,and angular momentum bias,and the applications of nonreciprocal thermal metamaterials are outlined.We also discuss nonreciprocal thermal radiation.Moreover,the potential applications of nonreciprocity to other Laplacian physical fields are discussed.Finally,the prospects for advancing nonreciprocal thermal metamaterials are highlighted,including developments in device design and manufacturing techniques and machine learning-assisted material design.

Viologen-based flexible electrochromic devices

Electrochromic technology has gained significant attention in various fields such as displays,smart windows,biomedical monitoring,military camouflage,human-machine interaction,and electronic skin due to its ability to provide reversible and fast color changes under applied voltage.With the rapid development and increasing demand for flexible electronics,flexible electrochromic devices(FECDs)that offer smarter and more controllable light modulation hold great promise for practical applications.The electrochromic material(ECM)undergoing color changes during the electrochemical reactions is one of the key components in electrochromic devices.Among the ECMs,viologens,a family of organic small molecules with 1,1'-disubstituted-4,4'-dipyridinium salts,have garnered extensive research interest,due to their well-reversible redox reactions,excellent electron acceptance ability,and the ability to produce multiple colors.Notably,viologen-based FECDs demonstrate color changes in the liquid or semisolid electrolyte layer,eliminating the need for two solid electrodes and thus simplifying the device structure.Consequently,viologens offer significant potential for the development of FECDs with high optical contrast,fast response speed,and excellent stability.This review aims to provide a comprehensive overview of the progress and perspectives of viologen-based FECDs.It begins by summarizing the typical structure and recent exciting developments in viologen-based FECDs,along with their advantages and disadvantages.Furthermore,the review discusses recent advancements in FECDs with additional functionalities such as sensing,photochromism,and energy storage.Finally,the remaining challenges and potential research directions for the future of viologen-based FECDs are addressed.

Exercise-induced adaptation of neurons in the vertebrate locomotor system

Vertebrate neurons are highly dynamic cells that undergo several alterations in their functioning and physiologies in adaptation to various external stimuli.In particular,how these neurons respond to physical exercise has long been an area of active research.Studies of the vertebrate locomotor system’s adaptability suggest multiple mechanisms are involved in the regulation of neuronal activity and properties during exercise.In this brief review,we highlight recent results and insights from the field with a focus on the following mechanisms:(a)alterations in neuronal excitability during acute exercise;(b)alterations in neuronal excitability after chronic exercise;(c)exercise-induced changes in neuronal membrane properties via modulation of ion channel activity;(d)exercise-enhanced dendritic plasticity;and(e)exercise-induced alterations in neuronal gene expression and protein synthesis.Our hope is to update the community with a cellular and molecular understanding of the recent mechanisms underlying the adaptability of the vertebrate locomotor system in response to both acute and chronic physical exercise.

A historical overview of nano-optics:From near-field optics to plasmonics

Nano-optics is an emergent research field in physics that appeared in the 1980s,which deals with light–matter optical interactions at the nanometer scale.In early studies of nano-optics,the main concern focus is to obtain higher optical resolution over the diffraction limit.The researches of near-field imaging and spectroscopy based on scanning near-field optical microscopy(SNOM)are developed.The exploration of improving SNOM probe for near-field detection leads to the emergence of surface plasmons.In the sense of resolution and wider application,there has been a significant transition from seeking higher resolution microscopy to plasmonic near-field modulations in the nano-optics community during the nano-optic development.Nowadays,studies of nano-optics prefer the investigation of plasmonics in different material systems.In this article,the history of the development of near-field optics is briefly reviewed.The difficulties of conventional SNOM to achieve higher resolution are discussed.As an alternative solution,surface plasmons have shown the advantages of higher resolution,wider application,and flexible nano-optical modulation for new devices.The typical studies in different periods are introduced and characteristics of nano-optics in each stage are analyzed.In this way,the evolution progress from near-field optics to plasmonics of nano-optics research is presented.The future development of nano-optics is discussed then.

Bile acids,gut microbiota,and therapeutic insights in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is a prevalent and aggressive liver malignancy.The interplay between bile acids(BAs)and the gut microbiota has emerged as a critical factor in HCC development and progression.Under normal conditions,BA metabolism is tightly regulated through a bidirectional interplay between gut microorganisms and BAs.The gut microbiota plays a critical role in BA metabolism,and BAs are endogenous signaling molecules that help maintain liver and intestinal homeostasis.Of note,dysbiotic changes in the gut microbiota during pathogenesis and cancer development can disrupt BA homeostasis,thereby leading to liver inflammation and fibrosis,and ultimately contributing to HCC development.Therefore,understanding the intricate interplay between BAs and the gut microbiota is crucial for elucidating the mechanisms underlying hepatocarcinogenesis.In this review,we comprehensively explore the roles and functions of BA metabolism,with a focus on the interactions between BAs and gut microorganisms in HCC.Additionally,therapeutic strategies targeting BA metabolism and the gut microbiota are discussed,including the use of BA agonists/antagonists,probiotic/prebiotic and dietary interventions,fecal microbiota transplantation,and engineered bacteria.In summary,understanding the complex BA-microbiota crosstalk can provide valuable insights into HCC development and facilitate the development of innovative therapeutic approaches for liver malignancy.

Systemic modulation of skeletal mineralization by magnesium implant promoting fracture healing: Radiological exploration enhanced with PCA-based machine learning in a rat femoral model

The clinical application of magnesium(Mg)and its alloys for bone fractures has been well supported by in vitro and in vivo trials.However,there were studies indicating negative effects of high dose Mg intake and sustained local release of Mg ions on bone metabolism or repair,which should not be ignored when developing Mg-based implants.Thus,it remains necessary to assess the biological effects of Mg implants in animal models relevant to clinical treatment modalities.The primary purpose of this study was to validate the beneficial effects of intramedullary Mg implants on the healing outcome of femoral fractures in a modified rat model.In addition,the mineralization parameters at multiple anatomical sites were evaluated,to investigate their association with healing outcome and potential clinical applications.Compared to the control group without Mg implantation,postoperative imaging at week 12 demonstrated better healing outcomes in the Mg group,with more stable unions in 3D analysis and high-mineralized bridging in 2D evaluation.The bone tissue mineral density(TMD)was higher in the Mg group at the non-operated femur and lumbar vertebra,while no differences between groups were identified regarding the bone tissue volume(TV),TMD and bone mineral content(BMC)in humerus.In the surgical femur,the Mg group presented higher TMD,but lower TV and BMC in the distal metaphyseal region,as well as reduced BMC at the osteotomy site.Principal component analysis(PCA)-based machine learning revealed that by selecting clinically relevant parameters,radiological markers could be constructed for differentiation of healing outcomes,with better performance than 2D scoring.The study provides insights and preclinical evidence for the rational investigation of bioactive materials,the identification of potential adverse effects,and the promotion of diagnostic capabilities for fracture healing.