All-in-One Alkaline Phosphatase-Response Aggregation-Induced Emission Probe for Cancer Discriminative Imaging and Combinational Chemodynamic-Photodynamic Therapy.

Currently, specific cancer-responsive fluorogenic probes with activatable imaging and therapeutic functionalities are in great demand in the accurate diagnostics and efficient therapy of malignancies. Herein, an all-in-one strategy is presented to realize fluorescence (FL) imaging-guided and synergetic chemodynamic-photodynamic cancer therapy by using a multifunctional alkaline phosphatase (ALP)-response aggregation-induced emission (AIE) probe, TPE-APP. By responding to the abnormal expression levels of an ALP biomarker in cancer cells, the phosphate groups on the AIE probe are selectively hydrolyzed, accompanied by in situ formation of strong emissive AIE aggregates for discriminative cancer cell imaging over normal cells and highly active quinone methide species with robust chemodynamic-photodynamic activities. Consequently, the activated AIE probes can efficiently destroy cancer cell membranes and lead to the death of cancer cells within 30 min. A superior efficacy in cancer cell ablation is demonstrated in vitro and in vivo. The cancer-associated biomarker response-derived discriminative FL imaging and synergistic chemodynamic-photodynamic therapy are expected to provide a promising avenue for precise image-guided cancer therapy.

Synchronously Sensitive Immunoassay and Efficient Inactivation of Living Zika Virus via DNAzyme Catalytic Amplification and In Situ Aggregation-Induced Emission Photosensitizer Generation.

Sensitive identification and effective inactivation of the virus are paramount for the early diagnosis and treatment of viral infections to prevent the risk of secondary transmission of viruses in the environment. Herein, we developed a novel two-step fluorescence immunoassay using antibody/streptavidin dual-labeled polystyrene nanobeads and biotin-labeled G-quadruplex/hemin DNAzymes with peroxidase-mimicking activity for sensitive quantitation and efficient inactivation of living Zika virus (ZIKV). The dual-labeled nanobeads can specifically bind ZIKV through E protein targeting and simultaneously accumulate DNAzymes, leading to the catalytic oxidation of Amplex Red indicators and generation of intensified aggregation-induced emission fluorescence signals, with a detection limit down to 66.3 PFU/mL and 100% accuracy. Furthermore, robust reactive oxygen species generated in situ by oxidized Amplex Red upon irradiation can completely kill the virus. This sensitive and efficient detection-inactivation integrated system will expand the viral diagnostic tools and reduce the risk of virus transmission in the environment.

A biocompatible dual-AIEgen system without spectral overlap for quantitation of microbial viability and monitoring of biofilm formation.

The lack of rapid and reliable microbial detection and sensing platforms and insufficient understanding of microbial behavior may delay precautions that could be made, which is a great threat to human life and increases the heavy financial burden on society. In this contribution, a dual-aggregation-induced emission luminogen (AIEgen) system is successfully developed for microbial imaging and metabolic status sensing. This system consists of two AIEgens (DCQA and TPE-2BA) that bear positively charged groups or boronic acid groups, providing universal microbial staining ability and specific affinity for dead microbes, respectively. Based on the distinctive fluorescence response produced by the diverse interaction of AIEgens with live or dead microbes, this dual-AIEgen system can detect all the microbes and identify their viabilities. Furthermore, the morphology and metabolic status of a sessile biofilm can also be imaged and monitored. The system exhibits rapid labelling properties that suitable for various microbes, and good biocompatibilities.

Ultrasensitive Visualization of Virus via Explosive Catalysis of an Enzyme Muster Triggering Gold Nano-aggregate Disassembly.

Sensitive and accurate diagnosis of viral infection is important for human health and social safety. Herein, by means of explosive catalysis from an enzyme muster, a powerful naked-eye readout platform has been successfully constructed for ultrasensitive immunoassay of viral entities. Liposomes were used to encapsulate multiple enzymes into an active unit. In addition, its triggered rupture could boost the disassembly of gold nano-aggregates that were cross-linked by peptides with opposite charges. As a result, plasmonically colorimetric signals were rapidly generated for naked-eye observation. Further harnessing the immunocapture, enterovirus 71 (EV71), a class of highly infective virus, was sensitively assayed with a detection limit down to 16 copies/µL. It is superior to the single enzyme-anchored immunoassay system. Most importantly, the colorimetric assay was demonstrated with 100% clinical accuracy, displaying strong anti-interference capability. It is expectable that this sensitive, accurate, and convenient strategy could provide a prospective alternative for viral infection analysis, especially in resource-constrained settings.

Less is more: Silver-AIE core@shell nanoparticles for multimodality cancer imaging and synergistic therapy.

Nanomaterials with integrated multiple imaging and therapeutic modalities possess great potentials in accurate cancer diagnostics and enhanced therapeutic efficacy. Traditional strategies for achieving multimodality nanoplatform through one by one combination of different modalities are challenged by the complicated structural design and fabrication as well as inherent incompatibility between different modalities. Herein, a novel strategy is presented to realize multimodal imaging and synergistic therapy using a class of simple silver core/AIE (aggregation-induced emission) shell nanoparticles. In addition to the intrinsic AIE fluorescence (FL) and metal-based computed tomography (CT) and radiation therapy (RT) properties, an extra functionality at the core/shell interface was identified to enable excellent photothermal (PT) and photoacoustic (PA) performance. As a result, five imaging and therapy modalities (FL, CT, PA, photothermal therapy (PTT), and RT) were achieved with a single structural unit for sensitive tumor imaging and effective therapy.

Three-Component Regio- and Stereoselective Polymerizations toward Functional Chalcogen-Rich Polymers with AIE-Activities.

Polymers containing rich chalcogen elements are rarely reported due to the lack of facile synthesis methods. Herein, a novel multicomponent polymerization route toward chalcogen-rich polymers was introduced. A series of poly(vinyl sulfones) (PVSs) were synthesized at room temperature using readily prepared monomers. PVSs were generated with high regio- and stereo-selectivity in high yields (up to 92.3%). Rich chalcogen elements endowed PVSs with distingctive multifunctionalities. The PVSs possessed good solubility and film-forming ability. Their thin films exhibited outstanding refractive indices up to 1.8062 at 550.0 nm together with good optical transparency in the visible region. Thin films of some polymers can also be fabricated into well-resolved fluorescent photopatterns by photolithography. Thanks to the unique redox properties of selenium, postmodification by oxidation reaction of P1a/2/3a successfully eliminates the caused heavy atom effect and endow resulting polymers with novel functionality as fluorescent bioprobes for cellular imaging.

AIE-based theranostic systems for detection and killing of pathogens.

Pathogenic bacteria, fungi and viruses pose serious threats to the human health under appropriate conditions. There are many rapid and sensitive approaches have been developed for identification and quantification of specific pathogens, but many challenges still exist. Culture/colony counting and polymerase chain reaction are the classical methods used for pathogen detection, but their operations are time-consuming and laborious. On the other hand, the emergence and rapid spread of multidrug-resistant pathogens is another global threat. It is thus of utmost urgency to develop new therapeutic agents or strategies. Luminogens with aggregation-induced emission (AIEgens) and their derived supramolecular systems with unique optical properties have been developed as fluorescent probes for turn-on sensing of pathogens with high sensitivity and specificity. In addition, AIE-based supramolecular nanostructures exhibit excellent photodynamic inactivation (PDI) activity in aggregate, offering great potential for not only light-up diagnosis of pathogen, but also image-guided PDI therapy for pathogenic infection.

Multifunctional Cellular Beacons with in Situ Synthesized Quantum Dots Make Pathogen Detectable with the Naked Eye.

The rapid and sensitive detection of pathogens is extremely crucial for timely clinical diagnosis and diseases control. Here, by employing cellular beacons with in situ synthesized QDs created from Staphylococcus aureus ( S. aureus), we efficiently fabricated an antibody (Ab) and acetylcholinesterase (AChE)-functionalized nanobioprobe, i.e., multifunctional cellular beacons (MCBs), avoiding complicated modification. Coupled with magnetic separation, a novel method for pathogen detection with the naked eye is established. With this method, enterovirus 71 (EV71) can be detected by the naked eye through the aggregation of gold nanoparticles that is triggered by the product of AChE catalyzed acetylthiocholine, with a detection limit of 0.5 ng/mL. Moreover, due to the MCBs have high luminance with perfect uniformity, the detection can also be realized by counting the number of MCBs, with a detection limit of 1 ng/mL. The method is validated with human throat swabs, resulting in a complete consistence with reverse transcription-polymerase chain reaction results. This study reports the first cellular beacons-based method for pathogen detection by the naked eye and broadens the applicability of cell self-synthesized nanoparticles-based immunoassays. Moreover, the MCBs-based method will provide a powerful tool for clinical detection.

AIE Featured Inorganic-Organic Core@Shell Nanoparticles for High-Efficiency siRNA Delivery and Real-Time Monitoring.

RNA interference (RNAi) is demonstrated as one of the most powerful technologies for sequence-specific suppression of genes in disease therapeutics. Exploration of novel vehicles for small interfering RNA (siRNA) delivery with high efficiency, low cytotoxicity, and self-monitoring functionality is persistently pursued. Herein, by taking advantage of aggregation-induced emission luminogen (AIEgen), we developed a novel class of Ag@AIE core@shell nanocarriers with regulable and uniform morphology. It presented excellent efficiencies in siRNA delivery, target gene knockdown, and cancer cell inhibition in vitro. What's more, an anticancer efficacy up to 75% was achieved in small animal experiments without obvious toxicity. Attributing to the unique AIE properties, real-time intracellular tracking of siRNA delivery and long-term tumor tissue imaging were successfully realized. Compared to the commercial transfection reagents, significant improvements were obtained in biocompatibility, delivery efficiency, and reproducibility, representing a promising future of this nanocarrier in RNAi-related cancer therapeutics.

Ultrasensitive Virion Immunoassay Platform with Dual-Modality Based on a Multifunctional Aggregation-Induced Emission Luminogen.

Sensitive and accurate detection of highly contagious virus is urgently demanded for disease diagnosis and treatment. Herein, based on a multifunctional aggregation-induced emission luminogen (AIEgen), a dual-modality readout immunoassay platform for ultrasensitive detection of viruses has been successfully demonstrated. The platform is relied on virions immuno-bridged enzymatic hydrolysis of AIEgen, accompanying with the in situ formation of highly emissive AIE aggregates and shelling of silver on gold nanoparticles. As a result, robust turn-on fluorescence and naked-eye discernible plasmonic colorimetry composed dual-signal is achieved. By further taking advantage of effective immunomagnetic enrichment, EV71 virions, as an example, can be specifically detected with a limit of detection down to 1.4 copies/µL under fluorescence modality. Additionally, semiquantitative discerning of EV71 virions is realized in a broad range from 1.3 × 103 to 2.5 × 106 copies/µL with the naked eye. Most importantly, EV71 virions in 24 real clinical samples are successfully diagnosed with 100% accuracy. Comparing to the gold standard polymerase chain reaction (PCR) assay, our immunoassay platform do not need complicated sample pretreatment and expensive instruments. This dual-modality strategy builds a good capability for both colorimetry based convenient preliminary screening and fluorescence based accurate diagnosis of suspect infections in virus-stricken areas.

Redox-Active AIEgen-Derived Plasmonic and Fluorescent Core@Shell Nanoparticles for Multimodality Bioimaging.

Multimodality imaging is highly desirable for accurate diagnosis by achieving high sensitivity, spatial-temporal resolution, and penetration depth with a single structural unit. However, it is still challenging to integrate fluorescent and plasmonic modalities into a single structure, as they are naturally incompatible because of significant fluorescence quenching by plasmonic noble-metal nanoparticles. Herein, we report a new type of silver@AIEgen (aggregation-induced emission luminogen) core-shell nanoparticle (AACSN) with both strong aggregated-state fluorescence of the AIEgen and distinctive plasmonic scattering of silver nanoparticles for multimodality imaging in living cells and small animals. The AACSNs were prepared through a redox reaction between silver ions and a redox-active AIEgen, which promoted synergistic formation of the silver core and self-assembly of the AIEgen around the core. The resulting AACSNs exhibited good biocompatibility and high resistance to environmental damage. As a result, excellent performance in fluorescence imaging, dark-field microscopy, and X-ray computed tomography-based multimodality imaging was achieved.

Highly Sensitive Naked-Eye Assay for Enterovirus 71 Detection Based on Catalytic Nanoparticle Aggregation and Immunomagnetic Amplification.

Development of sensitive, convenient, and cost-effective virus detection product is of great significance to meet the growing demand of clinical diagnosis at the early stage of virus infection. Herein, a naked-eye readout of immunoassay by means of virion bridged catalase-mediated in situ reduction of gold ions and growth of nanoparticles, has been successfully proposed for rapid visual detection of Enterovirus 71 (EV71). Through tailoring the morphologies of the produced gold nanoparticles (GNPs) varying between dispersion and aggregation, a distinguishing color changing was ready for observation. This colorimetric detection assay, by further orchestrating the efficient magnetic enrichment and the high catalytic activity of enzyme, is managed to realize highly sensitive detection of EV71 virions with the limit of detection (LOD) down to 0.65 ng/mL. Our proposed method showed a much lower LOD value than the commercial ELISA for EV71 virion detection. Comparing to the current clinical gold standard polymerase chain reaction (PCR) method, our strategy provided the same diagnostic outcomes after testing real clinical samples. Besides, this strategy has no need of complicated sample pretreatment or expensive instruments. Our presented naked-eye immunoassay method holds a promising prospect for the early detection of virus-infectious disease especially in resource-constrained settings.

Full-Genome Sequences of Seven Fatal Enterovirus 71 Strains Isolated in Shenzhen, China, in 2014.

The whole-genome sequences of seven fatal enterovirus 71 (EV71) strains, isolated in southern China, in 2014, were determined. The complete genome sequences of these strains displayed close relationships to native EV71 strains and showed 94.2% to 99.8% identity to each other. All of these strains were assigned to subgenotype C4a based on phylogenetic analysis of the VP1 gene.

Fluorescence-Converging Carbon Nanodots-Hybridized Silica Nanosphere.

Ultrabright carbon nanodots-hybridized silica nanospheres (CSNs) are synthesized through the Stöber process of silane functionalized C-dots. The fluorescence of carbon nanodots is converged intensely. A CSN is about 3800 times brighter than a single-carbon nanodot. Along with their high brightness and low cytotoxicity, CSNs also indicate their potential application in cellular labeling.

Visual and efficient immunosensor technique for advancing biomedical applications of quantum dots on Salmonella detection and isolation.

It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here by applying fluorescent nanobioprobes on a specially-designed cellulose-based swab (a solid-phase enrichment system). The selective and chromogenic medium used on this swab can achieve the ultrasensitive amplification of target bacteria and form chromogenic colonies in situ based on a simple biochemical reaction. More importantly, because this swab can serve as an attachment site for the targeted pathogens to immobilize and immunologically capture nanobioprobes, our mAb-conjugated QD bioprobes were successfully applied on the solid-phase enrichment system to capture the fluorescence of targeted colonies under a designed excitation light instrument based on blue light-emitting diodes combined with stereomicroscopy or laser scanning confocal microscopy. Compared with the traditional methods using 4-7 days to isolate Salmonella from the bacterial mixture, this method took only 2 days to do this, and the process of initial screening and preliminary diagnosis can be completed in only one and a half days. Furthermore, the limit of detection can reach as low as 10(1) cells per mL Salmonella on the background of 10(5) cells per mL non-Salmonella (Escherichia coli, Proteus mirabilis or Citrobacter freundii, respectively) in experimental samples, and even in human anal ones. The visual and efficient immunosensor technique may be proved to be a favorable alternative for screening and isolating Salmonella in a large number of samples related to public health surveillance.

Harnessing Intracellular Biochemical Pathways for In Vitro Synthesis of Designer Tellurium Nanorods.

Synthesizing nanomaterials of desired properties is a big challenge, which requires extremely harsh conditions and/or use of toxic materials. More recently developed in vivo methods have brought a different set of problems such as separation and purification of nanomaterials made in vivo. Here, a novel approach that harnesses cellular pathways for in vitro synthesis of high-quality tellurium nanorods with tunable lengths and optical properties is reported. It is first demonstrated that in vivo biochemical pathways could be used to synthesize Te nanorods via the intracellular reduction of TeO3(2-) in living Staphylococcus aureus cells. The pathways to set up a quasi-biological system for Te precursor formation are then utilized, which could further synthesize Te nanorods in vitro. This allows to successfully synthesize in vitro, under routine laboratory conditions, Te nanorods with uniform and tunable lengths, ranging from about 10 to 200 nm, and controllable optical properties with high molar extinction coefficients. The approach here should open new avenues for controllable, facile, and efficient synthesis of designer nanomaterials for diverse industrial and biomedical applications.

Uniform fluorescent nanobioprobes for pathogen detection.

Manipulating biochemical reactions in living cells to synthesize nanomaterials is an attractive strategy to realize their synthesis that cannot take place in nature. Yeast cells have been skillfully utilized to produce desired nanoparticles through spatiotemporal coupling of intracellular nonrelated biochemical reaction pathways for formation of fluorescent CdSe quantum dots. Here, we have successfully transformed Staphylococcus aureus cells into cellular beacons (fluorescing cells), all of which are highly fluorescent and photostable with perfect uniformity. Importantly, on the basis of such cells, we efficiently fabricated fluorescent nanobioprobes by a specific interaction between the protein A expressed on the S. aureus surface and the Fc fragment domain of antibodies, avoiding the use of other common methods for cell surface modifications, such as molecular covalent connection or more difficult genetic and metabolic engineering. Coupled with immunomagnetic beads, the resulting fluorescent-biotargeting bifunctional cells, i.e., biotargeting cellular beacons, can be employed as nanobioprobes for detection of viruses, bacteria, and tumor cells. With this method, H9N2 AIV can be detected specifically with a limit of 8.94 ng/mL (based on protein content). Furthermore, diverse probes for detection of different pathogens or for other biomedical applications can be easily obtained by simply changing the antibody conjugated to the cell surface.

Nanomechanical analysis of yeast cells in CdSe quantum dot biosynthesis.

QD biosynthesis affects the mechanical strength of yeast cells. The intracellular synthesis of CdSe QD in yeast cells incubated with Na2 SeO3 and subsequently with CdCl2 increases the glucan content of their cell walls, resulting in their enhanced mechanical strength.

An ultra-high sensitive platform for fluorescence detection of micrococcal nuclease based on graphene oxide.

Micrococcal nuclease (MNase) is the extracellular nuclease of Staphylococcus aureus (S. aureus). It preferentially digests single-stranded nucleic acids. The existence of MNase can be the standard to identify S. aureus and the content of MNase can be used to evaluate the pathogenicity of S. aureus. Herein, an ultra-high sensitive and selective fluorescent sensing platform for MNase is developed based on MNase-induced DNA strand scission and the difference in affinity of graphene oxide (GO) for single-stranded DNA containing different numbers of bases in length. In the absence of MNase, the adsorption of the dye-labeled ssDNA on GO makes the dyes close proximity to GO surface resulting in high efficiency quenching of fluorescence of the dyes. Conversely, and very importantly, in the presence of MNase, it cleaves the dye-labeled ssDNA into small fragments. The introduction of GO into the sensing solution results in weak quenching of the fluorescence of the dyes due to the weak affinity of the short dye-labeled oligonuleotide fragment to GO, and the fluorescence intensity gradually increases with increasing concentration of MNase. MNase can be detected in a range of 8×10⁻5 to 1.6×10⁻³ unit/mL with a detection limit of 2.7×10⁻5 unit/mL and good selectivity. The detection limit is of two orders of magnitude lower than those reported fluorescence MNase assays. Moreover, when the GO-based biosensor is used in S. aureus sample assays, preeminent fluorescence signals are obtained, thus the platform of the GO-based biosensor can be used to detect MNase in real-world samples.