CRISPR/Cas13a-Responsive and RNA-Bridged DNA Hydrogel Capillary Sensor for Point-of-Care Detection of RNA.

Disease diagnostics and surveillance increasingly highlight the importance of portable, cost-effective, and sensitive point-of-care (POC) detection of nucleic acids. Here, we report a CRISPR/Cas13a-responsive and RNA-bridged DNA hydrogel capillary sensor for the direct and visual detection of specific RNA with high sensitivity. The capillary sensor was simply prepared by loading RNA-cross-linking DNA hydrogel film (∼0.2 mm ± 0.02 mm) at the end of a capillary. When CRISPR/Cas13a specifically recognizes the target RNA, the RNA bridge in the hydrogel film is cleaved by the trans-cleavage activity of CRISPR/Cas13a, increasing the permeability of the hydrogel film. Different concentrations of target RNA activate different amounts of Cas13a, cleaving different amounts of the RNA bridge in the hydrogel and causing corresponding changes in the permeability of the hydrogel. Therefore, samples containing different amounts of the target RNA travel to different distances in the capillary. Visual reading of the distance provides quantitative detection of the RNA target without the need for any nucleic acid amplification or auxiliary equipment. The technique was successfully used for the determination of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in clinical nasopharyngeal (NP) swab and saliva samples. Easily quantifiable distance using a ruler eliminates the need for any optical or electrochemical detection equipment, making this assay potentially useful for POC and on-site applications.

Advances in Polysaccharides for Cartilage Tissue Engineering Repair: A Review.

Cartilage repair has been a significant challenge in orthopedics that has not yet been fully resolved. Due to the absence of blood vessels and the almost cell-free nature of mature cartilage tissue, the limited ability to repair cartilage has resulted in significant socioeconomic pressures. Polysaccharide materials have recently been widely used for cartilage tissue repair due to their excellent cell loading, biocompatibility, and chemical modifiability. They also provide a suitable microenvironment for cartilage repair and regeneration. In this Review, we summarize the techniques used clinically for cartilage repair, focusing on polysaccharides, polysaccharides for cartilage repair, and the differences between these and other materials. In addition, we summarize the techniques of tissue engineering strategies for cartilage repair and provide an outlook on developing next-generation cartilage repair and regeneration materials from polysaccharides. This Review will provide theoretical guidance for developing polysaccharide-based cartilage repair and regeneration materials with clinical applications for cartilage tissue repair and regeneration.

The Pharmacological Efficacy of Baicalin in Inflammatory Diseases.

Baicalin is one of the most abundant flavonoids found in the dried roots of Scutellaria baicalensis Georgi (SBG) belonging to the genus Scutellaria. While baicalin is demonstrated to have anti-inflammatory, antiviral, antitumor, antibacterial, anticonvulsant, antioxidant, hepatoprotective, and neuroprotective effects, its low hydrophilicity and lipophilicity limit the bioavailability and pharmacological functions. Therefore, an in-depth study of baicalin's bioavailability and pharmacokinetics contributes to laying the theoretical foundation for applied research in disease treatment. In this view, the physicochemical properties and anti-inflammatory activity of baicalin are summarized in terms of bioavailability, drug interaction, and inflammatory conditions.

Prospects of Using Chitosan-Based Biopolymers in the Treatment of Peripheral Nerve Injuries.

Peripheral nerve injuries are common neurological disorders, and the available treatment options, such as conservative management and surgical repair, often yield limited results. However, there is growing interest in the potential of using chitosan-based biopolymers as a novel therapeutic approach to treating these injuries. Chitosan-based biopolymers possess unique characteristics, including biocompatibility, biodegradability, and the ability to stimulate cell proliferation, making them highly suitable for repairing nerve defects and promoting nerve regeneration and functional recovery. Furthermore, these biopolymers can be utilized in drug delivery systems to control the release of therapeutic agents and facilitate the growth of nerve cells. This comprehensive review focuses on the latest advancements in utilizing chitosan-based biopolymers for peripheral nerve regeneration. By harnessing the potential of chitosan-based biopolymers, we can pave the way for innovative treatment strategies that significantly improve the outcomes of peripheral nerve injury repair, offering renewed hope and better prospects for patients in need.

A Network Pharmacology and Multi-Omics Combination Approach to Reveal the Effect of Strontium on Ca2+ Metabolism in Bovine Rumen Epithelial Cells.

Strontium (Sr) belongs to the same group in the periodic table as calcium (Ca). Sr level can serve as an index of rumen Ca absorption capacity; however, the effects of Sr on Ca2+ metabolism are unclear. This study aims to investigate the effect of Sr on Ca2+ metabolism in bovine rumen epithelial cells. The bovine rumen epithelial cells were isolated from the rumen of newborn Holstein male calves (n = 3, 1 day old, 38.0 ± 2.8 kg, fasting). The half maximal inhibitory concentration (IC50) of Sr-treated bovine rumen epithelial cells and cell cycle were used to establish the Sr treatment model. Transcriptomics, proteomics, and network pharmacology were conducted to investigate the core targets of Sr-mediated regulation of Ca2+ metabolism in bovine rumen epithelial cells. The data of transcriptomics and proteomics were analyzed using bioinformatic analysis (Gene Ontology and Kyoto Encyclopedia of genes/protein). Quantitative data were analyzed using one-way ANOVA in GraphPad Prism 8.4.3 and the Shapiro-Wilk test was used for the normality test. Results presented that the IC50 of Sr treatment bovine rumen epithelial cells for 24 h was 43.21 mmol/L, and Sr increased intracellular Ca2+ levels. Multi-omics results demonstrated the differential expression of 770 mRNAs and 2436 proteins after Sr treatment; network pharmacology and reverse transcriptase polymerase chain reaction (RT-PCR) revealed Adenosylhomocysteine hydrolase-like protein 2 (AHCYL2), Semaphoring 3A (SEMA3A), Parathyroid hormone-related protein (PTHLH), Transforming growth factor ß2 (TGF-ß2), and Cholesterol side-chain cleavage enzyme (CYP11A1) as potential targets for Sr-mediated Ca2+ metabolism regulation. Together these results will improve the current comprehension of the regulatory effect of Sr on Ca2+ metabolism and pave a theoretical basis for Sr application in bovine hypocalcemia.

Ultra-Trace Protein Detection by Integrating Lateral Flow Biosensor with Ultrasound Enrichment.

Lateral flow biosensor (LFB) is one of the most successful and applied commercial detection methods for food safety, drug abuse, and disease. Here, we integrated the ultrasound enrichment as sample preparation with LFB to achieve the ultra-trace protein detection in blood. When the ultrasound field is applied, the interaction between the acoustic field and gold nanoparticles can gather specifically modified gold nanoparticles toward pressure nodes in seconds and enrich target proteins. Such an approach can detect protein with a linear range of 1-20 ng mL-1 and detection limit of 0.58 ng mL-1 in blood within 20 min, which enormously reduces false positive readings caused by interference in real blood samples with complex components. Such a microchip that integrated acoustic enrichment with LFB shows great potential in detecting ultra-trace biomarkers for clinical diagnosis.

Characterizing microring resonators using optical frequency domain reflectometry.

A novel, to the best of our knowledge, method to extract optical microring resonators' loss characteristics is proposed and demonstrated using optical frequency domain reflectometry (OFDR). Compared with the traditional optical transmission measurement method, the spatial-resolved backscattering optical signals obtained from the OFDR can clearly show the resonance mode's increased optical path length due to its circulation inside the resonator. By further processing the backscattered optical signals, loaded $Q$-factors of several resonators can be accurately determined. A calculation model is proposed to derive the resonance mode's intrinsic $Q$-factor from OFDR measurements of a series of loaded resonators.

Core@Satellite Janus Nanomotors with pH-Responsive Multi-phoretic Propulsion.

We report core@satellite Janus mesoporous silica-Pt@Au (JMPA) nanomotors with pH-responsive multi-phoretic propulsion. The JMPA nanomotors first undergo self-diffusiophoretic propulsion in 3.0 % H2 O2 due to the isolation of the Au nanoparticles (AuNPs) from the PtNPs layer. Then the weak acidity of H2 O2 can trigger the disassembly and reassembly of the AuNPs, resulting in the Janus distribution of large AuNPs aggregates. Such reconstruction of JMPA leads to the contact between PtNPs and AuNPs aggregates, thus changing the propulsion mechanism to self-electrophoresis. The asymmetric and aggregated AuNPs also enable the generation of a thermal gradient under laser irradiation, which propels the JMPA nanomotors by self-thermophoresis. Such multi-phoretic propulsion offers considerable promise for developing advanced nanomachines with a stimuli-responsive switch of propulsion modes in biomedical applications.

Dual Stimuli-Responsive Controlled Release Nanocarrier for Multidrug Resistance Cancer Therapy.

Multidrug resistance of cancer cells is a major obstacle for cancer chemotherapy. Herein, we present a nanocarrier that can release chemotherapeutic agents to induce tumor cell death and generate NO under NIR to overcome multidrug resistance in cancer chemotherapy. Owing to the unique structure of the water channel in this controlled release system for chemotherapeutic agents, the nanocarrier surface is equipped with more active sites to graft NO donor molecules. The released NO performs very well in reversing multidrug resistance by inhibiting P-gp expression. Our findings provide new insight into multidrug resistance cancer therapy and controlled release nanocarriers for multiple drugs.

An indirect ELISA-inspired dual-channel fluorescent immunoassay based on MPA-capped CdTe/ZnS QDs.

To meet the need for high-throughput immunoassays, many multiplex fluorescent immunoassays have been proposed. Most of them need different kinds of fluorescent label indicators during the test. In this work, a novel indirect ELISA-inspired dual-channel fluorescent immunoassay based on 3-mercaptopropionic acid capped CdTe/ZnS quantum dots (QDs) was constructed. The ELISA wells were coated with two kinds of antigen-QD complex. When the primary antibodies were present in a sample, they mediated the binding of a secondary antibody-DNA-gold nanoparticle complex to the antigen-QD complex. Then the gold nanoparticles quenched the fluorescence of the QDs and a decrease in fluorescence intensity was observed. Thus, the amount of primary antibody could be estimated from the decrease of fluorescence intensity. Owing to the wide absorption range and the relatively narrow emission band of the QDs, the dual-channel fluorescent immunoassay system could work at the same excitation wavelength and the emission wavelengths of each channel had no interference. As a result, two different kinds of primary antibody could be detected at the same time in one ELISA well, which simplified the operation and greatly improved the efficiency. Besides, only one type of secondary antibody needs to be added to the prepared microtiter plates, which further simplified the operation during the detection procedure. This dual-channel fluorescent immunoassay system will provide new insights into high-throughput immunodetection. Graphical abstract.

Lateral flow biosensors based on the use of micro- and nanomaterials: a review on recent developments.

This review (with 187 refs.) summarizes the progress that has been made in the design of lateral flow biosensors (LFBs) based on the use of micro- and nano-materials. Following a short introduction into the field, a first section covers features related to the design of LFBs, with subsections on strip-based, cotton thread-based and vertical flow- and syringe-based LFBs. The next chapter summarizes methods for sample pretreatment, from simple method to membrane-based methods, pretreatment by magnetic methods to device-integrated sample preparation. Advances in flow control are treated next, with subsections on cross-flow strategies, delayed and controlled release and various other strategies. Detection conditionst and mathematical modelling are briefly introduced in the following chapter. A further chapter covers methods for reliability improvement, for example by adding other validation lines or adopting different detection methods. Signal readouts are summarized next, with subsections on color-based, luminescent, smartphone-based and SERS-based methods. A concluding section summarizes the current status and addresses challenges in future perspectives. Graphical abstractRecent development and breakthrough points of lateral flow biosensors.

Wettability alteration in a functional capillary tube for visual quantitative point of care testing.

Capillarity is an extremely common physical-chemical phenomenon related to wettability in nature, which has wide theoretical and practical interest. Herein, we reported a facile sensing device based on capillary force change in a vertical capillary tube. In this height-based capillary sensor (HCS), the inner surface of the capillary tube was modified with a layer of molecules with wetting responsibility based on the well-known simple surface chemistry. With targets in different concentrations, the wettability of the surface modified with responsive molecules would produce different changes. The responsive surfaces would change the capillary force of the vertical capillary tube, and result in different column heights. Like a thermometer, H+ and phenol have been quantified visually based on the height of the liquid inside the capillary tube.

Free-Blockage Mesoporous Anticancer Nanoparticles Based on ROS-Responsive Wetting Behavior of Nanopores.

To achieve an excellent delivery effect of drug, stimuli-responsive nano "gate" with physical blockage units is usually constructed on the surface of the mesoporous silica nanocarriers (MSNs). In nature, the aquaporins in cell membrane can control the transport of water molecules by regulating the channel wettability, which is resulted from the conformational change of amino acids in the channel. Inspired by this phonomenon, herein a new concept of free-blockage controlled release system is proposed, which is achieved by controlling the wettability of the internal surface of nanopores on MSNs. Such a new system is different from the physical-blockage controlled release system, which bypasses the use of nano "gate" and overcomes the limitations of traditional physical blockage system. Moreover, further studies have shown that the system can selectively release the entrapped doxorubicin in human breast adenocarcinoma (MCF-7) cells triggered by intracellular reactive oxygen species (ROS) but not in normalhuman umbilical vein endothelial cells (HUVECs) containing ROS with low levels. The wettability-determined free-blockage controlled release system is simple and effective, and it can also be triggered by intracellular biological stimuli, which provides a new approach for the future practical application of drug delivery and cancer therapy.

A Voltage-Responsive Free-Blockage Controlled-Release System Based on Hydrophobicity Switching.

Controlled-release systems based on mesoporous silica nanomaterials (MSNs) have drawn great attention owing to their potential biomedical applications. Various switches have been designed to control the release of cargoes through the construction of physical blocking units on the surface of MSNs. However, such physical blockages are limited by poor sealing ability and low biocompatibility, and most of them lack closure ability. Herein, a voltage-responsive controlled-release system was constructed by functionalizing the nanopore of MSNs with ferrocene. The system realized free-blockage controlled release and achieved pulsatile release. The nanopores of the ferrocene-functionalized MSNs were hydrophobic enough to prevent invasion of the solution. Once a suitable voltage was applied, the nanopores became hydrophilic, which was followed by invasion of the solution and the release of the cargos. Moreover, pulsatile release was realized, which avoided unexpected release after the stimulus disappeared. Thus, we believe that our studies provide new insight into highly effective blockage for MSNs. Furthermore, the voltage-responsive release system is expected to find use in electrical stimulation combination therapy and bioelectricity-responsive release.

Mechanistic Investigation of Visible-Light-Induced Intermolecular [2 + 2] Photocycloaddition Catalyzed with Chiral Thioxanthone.

The recent thioxanthone-sensitizer-catalyzed intermolecular [2 + 2] cycloaddition induced by visible-light irradiation set the stage for the future development of feasible photocycloadditions. Nonetheless, the mechanism of this reaction still remains under debate, especially on the activation mode of the thioxanthone photosensitizer (energy transfer, bielectron exchange, and hydrogen transfer are all possible mechanisms). To settle this issue, systematic density functional theory calculations have been carried out. The results indicate that the energy-transfer pathway is more favorable than the bielectron-exchange and the hydrogen-transfer pathways. Meanwhile, the overall transformations involve the complexation and excitation of photosensitizer, the first C-C bond formation, the dissociation of the sensitizer, the triplet-to-singlet electronic state crossing, and the second C-C bond formation. The first C-C bond formation is the rate- and selectivity-determining step, and synergistic energy and electron transfer from photosensitizer to substrate moieties takes place along this process. On this basis, the effect of olefin substrates (ethyl vinyl ketone vs vinyl acetate) on the stereoselectivity was finally analyzed.

Ultra-short FBG based distributed sensing using shifted optical Gaussian filters and microwave-network analysis.

Ultrashort fiber Bragg gratings (US-FBGs) have significant potential as weak grating sensors for distributed sensing, but the exploitation have been limited by their inherent broad spectra that are undesirable for most traditional wavelength measurements. To address this, we have recently introduced a new interrogation concept using shifted optical Gaussian filters (SOGF) which is well suitable for US-FBG measurements. Here, we apply it to demonstrate, for the first time, an US-FBG-based self-referencing distributed optical sensing technique, with the advantages of adjustable sensitivity and range, high-speed and wide-range (potentially >14000 µÎµ) intensity-based detection, and resistance to disturbance by nonuniform parameter distribution. The entire system is essentially based on a microwave network, which incorporates the SOGF with a fiber delay-line between the two arms. Differential detections of the cascaded US-FBGs are performed individually in the network time-domain response which can be obtained by analyzing its complex frequency response. Experimental results are presented and discussed using eight cascaded US-FBGs. A comprehensive numerical analysis is also conducted to assess the system performance, which shows that the use of US-FBGs instead of conventional weak FBGs could significantly improve the power budget and capacity of the distributed sensing system while maintaining the crosstalk level and intensity decay rate, providing a promising route for future sensing applications.

Radio-frequency unbalanced M-Z interferometer for wavelength interrogation of fiber Bragg grating sensors.

The optical unbalanced Mach-Zehnder interferometer (UMZI) has attracted significant interests for interrogation of FBG sensors owing to its excellent advantages in sensitivity, resolution, and demodulation speed. But this method is still limited to dynamic measurements due to its poor stability and reliability when used for quasi-static detections. Here, we propose for the first time, to the best of our knowledge, a radio-frequency unbalanced M-Z interferometer (RF-UMZI) for interrogation of FBG sensors, which, owing to its operation in an incoherent rather than a coherent regime, provides an ideal solution for the existing stability problem of the conventional UMZI, with remarkable features of adjustable resolution and potentially extremely high sensitivity. A dispersion compensation fiber (DCF) and single-mode fiber (SMF) with a small length difference are served as the two unbalanced arms of the RF interferometer. The induced differential chromatic dispersion transfers the wavelength shift of the FBG to the change of the RF phase difference between the two interferometric carriers, which ultimately leads to the variation of the RF signal intensity. An interrogation of a strain-turned FBG was accomplished and a maximum sensitivity of 0.00835 a.u./µÎµ was obtained, which can easily be further improved by more than two orders of magnitude through various fiber dispersion components. Finally, the stability of the interrogation was tested.

A three-line lateral flow biosensor for logic detection of microRNA based on Y-shaped junction DNA and target recycling amplification.

A rapid, sensitive, and accurate detection strategy for microRNA 16 (miR-16) was developed, which combined the convenience of lateral flow biosensors (LFBs), the design flexibility of Y-shaped junction DNA probe, and the enhancement ability of endonuclease-assisted target recycling amplification. The system is composed of a molecular beacon (MB) probe, an assistant probe, and endonuclease Nt.BbvCI, which plays the role of signal translation and amplification. In the presence of the target microRNAs (miRNAs), three chains of nucleic acid could hybridize with each other to form a Y-shaped junction structure, which could be recognized by the endonuclease Nt.BbvCI. The MB probe was efficiently cleaved by endonuclease and produced two new DNA fragments, while the regenerated assistant probe and target were hybridized to another MB probe and entered into the next cycle of the amplification. In this way, the detection of the readily biodegradable miRNA was turned into the detection of two DNA fragments in the LFB. Meanwhile, the detection of two different DNAs would improve the accuracy and effectively avoid false results. The amplified products containing DNA fragments were then applied to the lateral flow nucleic acid biosensor (LFNAB) with two test zones, on which specific DNA probes were designed. The formed DNA-DNA/gold nanoparticle (GNP) conjugates were captured and accumulated to produce two red bands in two test zones. The logic judgment of the two test zones provided more accurate and convincing results. Under optimal conditions, the visual detection limit of miR-16 in aqueous solutions was 0.1 pM, which is 100-1000 times lower than that of visual or colorimetric methods in the literature. It could be used for on-field and point-of-care testing and meet the urgent demand of sensitive and selective miRNA detection in remote rural areas without costly equipment. The system displayed good universality, compatibility, high specificity, and stability of miRNA detection, which revealed significant potentiality in biomedical diagnosis. Graphical abstract A simple and rapid approach for microRNA-16 (miR-16) detection was developed based on Y-shaped junction DNA probe and the endonuclease-assisted target recycling amplification using lateral flow biosensors.

Reverse-Bumpy-Ball-Type-Nanoreactor-Loaded Nylon Membranes as Peroxidase-Mimic Membrane Reactors for a Colorimetric Assay for H2O2.

Herein we report for the first time fabrication of reverse bumpy ball (RBB)-type-nanoreactor-based flexible peroxidase-mimic membrane reactors (MRs). The RBB-type nanoreactors with gold nanoparticles embedded in the inner walls of carbon shells were loaded on nylon membranes through a facile filtration approach. The as-prepared flexible catalytic membrane was studied as a peroxidase-mimic MR. It was found that the obtained peroxidase-mimic MR could exhibit several advantages over natural enzymes, such as facile and good recyclability, long-term stability and easy storage. Moreover, the RBB NS-modified nylon MRs as a peroxidase mimic provide a useful colorimetric assay for H2O2.

A free-blockage controlled release system based on the hydrophobic/hydrophilic conversion of mesoporous silica nanopores.

A pH-responsive free-blockage release system was achieved through controlling the hydrophobic/hydrophilic conversion of mesoporous silica nanopores. This system further presented pulsatile release with changing pH values between 4.0 and 7.0 for several cycles. This free-blockage release system could also release antitumor agents to induce cell death after infecting tumor cells and could have the ability of continuous infection to tumor cells with high drug-delivery efficiency and few side effects.