Arsenic trioxide (ATO) shows limited efficacy against solid tumours, largely because it induces protective autophagy that attenuates its pro-apoptotic activity. Our previous studies established that nanodiamonds (NDs) function as nanoparticle autophagy inhibitors (NAPIs) when delivered systemically, markedly enhancing ATO efficacy in orthotopic liver tumour models by blocking NUPR1-mediated autolysosomal clearance. However, systemic administration remains inefficient in modulating the local autophagic microenvironment within tumours. Here we developed an interventional strategy based on intratumoral injection to assess the feasibility and biosafety of locally blocking autophagic flux while substantially reducing the required ATO dosage, thereby maximising the synergistic anti-tumour effects of NDs and ATO. In HepG2 hepatocellular carcinoma cells, NDs markedly blocked the late stage of autophagic flux, thereby significantly amplifying ATO-induced apoptosis. In a subcutaneous xenograft liver cancer mouse model, intratumoral co-administration of NDs with low-dose ATO achieved ~91% tumour inhibition and effectively eliminated the systemic toxicity associated with high-dose ATO monotherapy. Notably, the synergistic antitumor effect was independent of increased intratumoral ATO accumulation and was driven instead by targeted modulation of the autophagic pathway. Collectively, this study demonstrates a mechanism of localised, nanomaterial-mediated autophagy regulation and offers an efficient, safe strategy for interventional therapy of advanced solid tumours.

Chiral crystalline porous materials demonstrate significant potential in enantiomeric separation, asymmetric catalysis, circularly polarized luminescence (CPL), and chiral sensing. However, constructing stable porous molecular crystals (PMCs) with well-defined chirality and permanent porosity remains challenging. Herein, we report the successful preparation of a pair of homochiral porous molecular crystals (PMCs), denoted as (R)-/(S)-BINAM-PMC, via ionic self-assembly of axially chiral 1,1′-binaphthyl-2,2′-diamine (BINAM) as a basic building block with 4,4′-biphenyldisulfonic acid (BPDS). The ionic pairs formed between sulfonate anions and ammonium cations assemble into one-dimensional (1D) chains through electrostatic interactions, which further extend into two-dimensional (2D) layered structures and ultimately stack into a three-dimensional (3D) porous framework with 1D channels. The material maintains structural and porous integrity after guest removal, demonstrating excellent stability and permanent porosity. Furthermore, circular dichroism (CD) spectroscopy confirms its distinct chiral nature. This work provides valuable insights for the design and fabrication of stable chiral crystalline porous materials.

Inspired by nature, the orchestration of self-assembling building blocks into hierarchical superstructures offers a transformative approach to functional materials design. While significant advances have been made in engineering solid-state hierarchical materials such as crystals and superlattices, creating dynamic, liquid-like hierarchical materials remains a profound challenge. Herein, a universal and efficient method is introduced to construct spherical nucleic acids (SNAs) functionalized with diverse nucleic acids (NAs), including random DNA sequences, circular DNA (circ-DNA), single guide RNA (sgRNA), messenger RNA (mRNA), and multi-branched DNA independent of sequence, length, or topology. By examining spatial configuration and mechanical rigidity in DNA-mediated bonding, precise hierarchical assembly of SNAs is enabled. Furthermore, using these multivalent SNAs as programmable molecule equivalents, liquid-phase hierarchical materials via phase separation are successfully created, forming microscale SNA droplets. These metal condensates exhibit dynamic liquid-like properties and stimuli-responsiveness, including enhanced photothermal effects in living cells. Our findings provide fundamental insights into the formation and dynamics of liquid hierarchical materials, offering potentials for designing living-matter-inspired systems and advancing applications in biomedicine and responsive materials.

Shanghai Synchrotron Radiation Facility (SSRF) is a third-generation 3.5 GeV synchrotron facility located on the Chinese mainland, operational for user applications since 2009. With the completion of its Phase II project this year, SSRF now supports over 40 experimental stations across various research fields. For the structural biology community, there are three macromolecular crystallography (MX) beamlines (BL02U1, BL17UM and BL10U2) and one endstation at the white X-ray beamline (BL03HB) managed by SSRF to meet the needs of both academic and industrial users seeking to determine macromolecular crystal structures. The MX group at SSRF is dedicated to continuously upgrading these beamlines in terms of technology and scientific capabilities. This paper reports on the current status of all the MX beamlines at SSRF and discusses emerging trends.

DNA has emerged as a promising medium for high-density and energy-efficient data storage, providing a promising solution to capacity constraints and energy demands resulting from the exponential growth of data. Long-term stable storage of DNA data in ambient conditions is a critical requirement for its practical application. Whereas strategies have been proposed to protect DNA from environmental degradation, they are often at the expense of data storage density. Here, we propose a strategy via interweaving metal-organic framework (MOF) and DNA to synthesize DNA-MOF crystal (DNA-MOF), a high-density and long-lifespan DNA-based information material. The uniform distribution of DNA molecules within the crystalline frameworks enables a high loading capacity of up to 30 wt%, corresponding to a theoretical information density of 106.6 exabytes/g. The crystal structure of MOF-DNA demonstrates efficient resistance to environmental damage such as heat, ultraviolet radiation, DNase, etc., achieving an estimated archival half-life over 24,000 years at 9.4 °C. The interwoven DNA-MOF crystals establish a material foundation for the reliable operation of DNA storage systems in open-field deployment and provide a new pathway toward sustainable long-term management of massive data. ### Competing Interest Statement The authors have declared no competing interest. National Key R&D Program of China, 2023YFA0915200 National Natural Science Foundation of China, T2188102, 22322704, U24A20497

Multivalent display of antigens can boost subunit vaccine immunogenicity. However, owing to the inherent difficulty in programmatically controlling the topology of multivalent antigens, its impact on antigen immunogenicity remains elusive. In this study, DNA-mediated modular precision assembly is employed to organize SARS-CoV-2 receptor-binding domains (RBDs) with different topological connections while preserving their epitopes. It is found that branching-connected RBDs induced significantly higher IgG titers than linear-connected RBDs at higher antigen valency (≥4). This increase in IgG response is associated with stronger B cell proliferation, likely due to enhanced antigen-receptor synergistic interactions leading to enhanced B cell receptor signaling. Branching-connected RBDs also provided superior humoral immunity in mice and stronger protection in SARS-CoV-2-infected hamsters compared to adjuvanted RBD. This work highlights the role of antigen topology in vaccine design and offers a universal modular platform for producing more effective subunit vaccines.

Despite the tremendous potential of the CRISPR/Cas9 gene-editing technology in precision therapeutics, intracellular delivery remains a major challenge. High cytoplasmic viscosity and lysosomal entrapment significantly impair the cytosolic transport and gene-editing efficiency. In this study, we demonstrate that both the size and magnetic responsiveness of Fe3O4 nanoclusters can be finely tuned by modulating ionic strength, enabling their rapid propulsion under external magnetic fields. Leveraging this property, we develop magnetic nanoparticle cluster nanorobots (MagCbots) of approximately 200 nm in size by electrostatically assembling Fe3O4 nanoclusters with CRISPR-Cas9 plasmids. Under magnetic actuation, MagCbots exhibit rapid rotation in highly viscous intracellular environments, achieving a linear velocity of ∼0.41 μm/s. MagCbots reduce intracellular viscosity by approximately 50% and enhance lysosomal escape efficiency by 3-fold compared to nonactuated counterparts. Their porous architecture not only offers high payload capacity but also protects plasmid DNA from enzymatic degradation. Notably, MagCbots enable efficient genome editing of both PD1 and PLK1 genes across various cell lines including hard-to-transfect Jurkat T cells. This magnetically driven nanorobot platform presents a promising strategy for active intracellular delivery and holds significant potential for advancing gene therapy and related biomedical applications.

DNA nanostructures, known for their programmability, ease of modification, and favourable biocompatibility, have gained widespread application in the biomedical field. Among them, Tetrahedral DNA Origami (TDOs), as a novel DNA nanostructure, possesses well-defined structures, multiple modification sites, and large cavities, making it a promising drug carrier. However, current understanding of TDOs' interactions with biological systems, particularly with target cells and organs, remains unexplored, limiting its further applications in biomedicine. In this work, we prepared TDOs with an average particle size of 40 nm and labelled them with Cy5 fluorescent molecules. Following intravenous injection in mice, the uptake of TDOs by different types of liver and kidney cells was observed. Results indicated that TDOs accumulate in renal tubules and are metabolized by Kupffer cells, epithelial cells, and hepatocytes in the liver. Additionally, in a tumour-bearing mouse model, TDOs passively targeted tumour tissues and exhibited excellent tumour penetration and retention after rapid metabolism in hepatocytes. Our findings provide crucial insights for the development of TDO-based drug delivery systems.

Crystalline porous organic salts (CPOSs), an emerging category of crystalline porous organic materials with potential applications in various fields, have garnered significant attention in recent years. However, the limited variety of building blocks, the non-directionality of ionic bonds, and the scarcity of design principles severely restrict the synthesis and widespread application of CPOSs, especially for two-dimensional (2D) CPOSs. Adopting suitable 2D building blocks is an effective way to construct 2D CPOSs with specific functions. Herein, a new CPOS (CPOS-9) with fluorescent properties has been synthesized by employing a four-node 2D fluorescent organic base and a linear two-node organic acid. Notably, CPOS-9 features a unique 2D layered structure formed by the aggregation of hydrophobic and hydrophilic groups, which stacks through electrostatic interactions rather than the conventional pi-pi stacking. With significant fluorescent characteristics and abundant binding sites within nanoconfined channels, CPOS-9 showcases high sensitivity and selectivity for Ce3+ detection, with a detection limit as low as 80 nM. Based on theoretical calculations, for the first time, an energy level matching mechanism is proposed to elucidate the fluorescence quenching observed during the detection of Ce3+ in porous organic materials. This work enriches the variety of 2D CPOSs and highlights their application in the detection of rare earth elements.

Synchrotron-based X-ray microscopy (XRM) has garnered widespread attention from researchers due to its high spatial resolution and excellent energy (element) resolution. Existing molecular probes suitable for XRM include immune probes and genetic labeling probes, enabling the precise imaging of various biological targets within cells. However, immune labeling techniques are prone to cross-interference between antigens and antibodies. Genetic labeling technologies have limited systems that allow express markers independently, and moreover, genetically encoded labels based on catalytic polymerization lack a fixed morphology. When applied to cell imaging, this can result in reduced localization accuracy due to the diffusion of labels within the cells. Therefore, both techniques face challenges in simultaneously labeling multiple biotargets within cells and achieving high-precision imaging. In this work, we applied the click reaction and developed a third category of imaging probes suitable for XRM, termed clickable X-ray nanoprobes (Click-XRN). Click-XRN consists of two components: an X-ray-sensitive multicolor imaging module and a particle-size-controllable morphology module. Efficient identification of intra- and extracellular biotargets is achieved through click reactions between the probe and biomolecules. Click-XRN possesses a controllable particle size, and its loading of various metal ions provides distinctive signals for imaging under XRM. Based on this, we optimized the imaging energy of Click-XRN with different particle sizes, enabling single-color and two-color imaging of the cell membrane, cell nucleus, and mitochondria with nanoscale spatial nanometers. Our work provides a potent molecular tool for investigating cellular activities through XRM.

An integrated computer software system for macromolecular crystallography (MX) data collection at the BL02U1 and BL10U2 beamlines of the Shanghai Synchrotron Radiation Facility is described. The system, Finback , implements a set of features designed for the automated MX beamlines, and is marked with a user-friendly web-based graphical user interface (GUI) for interactive data collection. The Finback client GUI can run on modern browsers and has been developed using several modern web technologies including WebSocket, WebGL, WebWorker and WebAssembly. Finback supports multiple concurrent sessions, so on-site and remote users can access the beamline simultaneously. Finback also cooperates with the deployed experimental data and information management system, the relevant experimental parameters and results are automatically deposited to a database.

"Drawing inspiration from nature" offers a wealth of creative possibilities for designing cutting-edge materials with improved properties and performance. Nature-inspired thylakoid-based nanoarchitectures, seamlessly integrate the inherent structures and functions of natural components with the diverse and controllable characteristics of nanotechnology. These innovative biomaterials have garnered significant attention for their potential in various biomedical applications. Thylakoids possess fundamental traits such as light harvesting, oxygen evolution, and photosynthesis. Through the integration of artificially fabricated nanostructures with distinct physical and chemical properties, novel photosynthetic nanoarchitectures can be catalytically generated, offering versatile functionalities for diverse biomedical applications. In this article, an overview of the properties and extraction methods of thylakoids are provided. Additionally, the recent advancements in the design, preparation, functions, and biomedical applications of a range of thylakoid-based photosynthetic nanoarchitectures are reviewed. Finally, the foreseeable challenges and future prospects in this field is discussed.

One of the basic goals of cell biology is to identify multiple biological molecules within cells and understand the complex interactions between biological molecules in cellular life activities. Synchrotron-based X-ray microscopy has high spatial resolution and good energy (element) resolution, which has great application potential in the recognition and imaging of intracellular biomolecules. At present, the probes that have been developed for synchrotron-based X-ray microscopy are mainly immunostaining probes and genetic labeling probes. Immunostaining probes are prone to lead to cross- reactions due to their dependence on antigen-antibody reactions. The genetic labeling probes, based on gene coding tags, catalyze the generation of X-ray sensitive polymers. However, the polymers used to provide X-ray imaging signals have no fixed morphology. When it is applied to cell imaging, the positioning accuracy will be reduced due to the diffusion of tags in cells, which is an inherent defect of such imaging tags. In addition, there are few existing systems that can express each other independently and step by step for this type of probe. Therefore, both types of X-ray probes mentioned above are difficult to achieve simultaneous high-resolution imaging observation of multiple biological target molecules in cellular life activities. In this research paper, by using the characteristics of X-ray that has good energy resolution and does not interfere with each other between element spectra, we can synthesize polydopamine (PDA) nanoparticles in a controlled manner, modify azide groups on PDA nanoparticles and chelate metal ions, develop a click chemistry based synchronous X-ray imaging tag (PDA-N3-Metal), and conduct synchrotron radition X-ray imaging on the tag with an imaging resolution of 30 nm. The research results lay a good foundation for further preparation of X-ray probes based on click chemistry, and for realizing the specific recognition and imaging of multiple biological molecules in cells at the same time.

DNA has been used as a robust material for the building of a variety of nanoscale structures and devices owing to its unique properties. Structural DNA nanotechnology has reported a wide range of applications including computing, photonics, synthetic biology, biosensing, bioimaging, and therapeutic delivery, among others. Nevertheless, the foundational goal of structural DNA nanotechnology is exploiting DNA molecules to build three-dimensional crystals as periodic molecular scaffolds to precisely align, obtain, or collect desired guest molecules. Over the past 30 years, a series of 3D DNA crystals have been rationally designed and developed. This review aims to showcase various 3D DNA crystals, their design, optimization, applications, and the crystallization conditions utilized. Additionally, the history of nucleic acid crystallography and potential future directions for 3D DNA crystals in the era of nanotechnology are discussed.

BL10U2 is an undulator-based macromolecular crystallography (MX) beamline located at the 3.5-GeV Shanghai Synchrotron Radiation Facility. BL10U2 is specifically designed for conducting routine and bio safety level-2 (BSL-2) MX experiments utilizing high-flux tunable X-rays with energies from 7 to 18 keV, providing a beam spot size of 20 µm (horizontal) × 10 µm (vertical) at the sample point. Certification by the Shanghai Pudong Municipal Health Commission confirmed the capability to perform BSL-2 MX experiments. The beamline is currently equipped with an Eiger X 16M detector and two newly developed in-house high-precision diffractometers that can be switched to perform conventional or in situ crystal diffraction experiments. An automatic sample changer developed in-house allows fast sample exchange in less than 30s, supporting high-throughput MX experimentation and rapid crystal screening. Data collection from both the diffractometer and detector was controlled by an in-house developed data collection software (Finback) with a user-friendly interface for convenient operation. This study presents a comprehensive overview of the facilities, experimental methods, and performance characteristics of the BL10U2 beamline.

Macromolecular crystallography beamline BL17U1 at the Shanghai Synchrotron Radiation Facility has been relocated, upgraded, and given a new ID (BL02U1). It now delivers X-rays in the energy range of 6–16 keV, with a focused beam of 11.6 µm × 4.8 µm and photon flux greater than 10 12 phs/s. The high credibility and stability of the beam and good timing synchronization of the equipment significantly improve the experimental efficiency. Since June 2021, when it officially opened to users, over 4200 h of beamtime have been provided to over 200 research groups to collect data at the beamline. Its good performance and stable operation have led to the resolution of several structures based on data collected at the beamline.

BackgroundSodium dodecyl sulfate (SDS) and polyethylene glycol octyl phenyl ether (Triton X-100) are commonly used for surfactants in the laboratory, which can dissolve lipids in the cell membrane, increase the permeability of cell membrane, and can assist the rapid penetration of staining reagents. Surfactants play an important role in immunostaining or tissue clearance.PurposeThis study aims to study the effect of SDS and Triton X-100 on cell membrane permeability.MethodsFirstly, the damage effects of SDS and Triton X-100 on cell membranes were compared and the optimal conditions were optimized. Then, the influence of the selection of SDS and Triton X-100 on the specific experimental results was investigated. Finally, the full-field transmission imaging technology of the soft X-ray nanoimaging beamline of the National Synchrotron Radiation Laboratory at Hefei was used to compare their affection on cell morphology before and after the cell rupture.ResultsExperimental results show that 0.25% Triton X-100 has better membrane penetration than 4% SDS, and the optimal treatment time of 0.25% Triton X-100 is 10 min. In the experiment of peroxidase catalyzed 3,3'-Diaminobenzidine (DAB) polymerization, SDS, rather than Triton X-100, affects the catalytic activity of enzymes, hence changes the staining effect. The synchrotron radiation high-resolution imaging demonstrate that there is no significant difference of cell morphology before and after the cell membrane permeability with Triton X-100.ConclusionsTriton X-100 is a better surfactant than SDS in the cell membrane permeability. The principal method of this study provides a good support for selecting suitable surfactants in the experiments.

Macromolecular crystallography is commonly used to determine the structure of biological macromolecules. Currently the beamlines at synchrotron radiation facilities play an important role in macromolecular crystallography, and have produced an enormous number of molecular structures to help solve scientific questions and support applications. Structure information makes significant contributions to the virus-related research as well. However, it is mandatory to be protected the operators under a compatible biosafety infrastructure when a pathological agent is set up in a beamline. Here a level-2 biosafety protection for a macromolecular crystallography beamline at Shanghai Synchrotron Radiation Facility (SSRF) is introduced. To fulfill the biosafety in a radioactive environment, a dedicated design is implemented. Since the beamline will be opened to the external users from nationwide research units, the management process and experimental method are also drawn up.