Wei Cheng

Electrochemical point‐of‐care testing for complex clinical samples is frequently hampered by matrix interference, signal instability, and laborious electrode modification procedures. Herein, we report a magnetic portable electrochemical immunosensor utilizing a conductive, anti‐fouling hydrogel coating composed of sodium alginate, polyethylene glycol, and g raphene (SPG). Upon calcium‐ion triggering, the SPG solution forms a stable electrode hydrogel coating within 1 min, effectively resisting nonspecific adsorption while maintaining low‑impedance electron transfer. Guided by a 3D‐printed magnetic apparatus, the MpECis directly captures target‐bound magnetic complexes, enabling rapid enrichment and electrical readout within 25 min. By implementing time‑domain averaging over the enzymatic reaction plateau, the coefficient of variation of electrical signals generated by nanoenzyme catalysis was reduced to 3.4%, markedly improving measurement reproducibility. The clinical practicality of the SPG MpECis platform was validated through three independent cohorts, including detection of cardiac troponin I in 59 serum samples, quantification of extracellular vesicles in 47 plasma samples, and detection of C‐reactive protein in 17 fingertip blood samples. Compared with the reference method, the area under the curve of this method are 0.99, 0.93, and 1.00, respectively. This platform offers a practical route for stable electrochemical diagnostics in complex biological matrices and home‐based precision medicine.

Aberrant proximal biomolecular complexes are critical disease biomarkers. The precise in situ imaging of these complexes is essential for deciphering disea

Preterm birth (PTB) is a leading cause of neonatal morbidity and mortality. Although increasing evidence links vaginal microbiota to preterm birth, clinica

Vitiligo pathogenesis involves progressive melanocyte loss and keratinocyte dysfunction, which are driven primarily by oxidative stress resulting from excessive ROS accumulation. We engineered a temporally controlled hydrogel microneedle system that integrates ginseng-derived exosomes (G-Exos) with biomimetic polydopamine nanoparticles (PDA@PEGs) to concurrently target the pathogenic triad of vitiligo, including oxidative stress, inflammation, and melanocyte deficiency. This system employs methacrylated hyaluronic acid (HAMA) hydrogel microneedles for rapid PDA@PEG release while utilizing glyceryl monostearate micelles to achieve matrix metalloproteinase-9 (MMP-9)-responsive G-Exo release at inflammatory foci, enabling intelligent spatiotemporal control. Functionally, G-Exos help restore redox homeostasis and suppress inflammation through bioactive constituents, thereby protecting melanocytes and enhancing keratinocyte proliferation. Moreover, PDA@PEG promotes repigmentation through the dual mechanisms of exogenous melanin deposition and endogenous melanogenesis stimulation. In murine models, this strategy achieves significant repigmentation within 3 weeks by activating follicular stem cells, upregulating melanogenic markers (Tyr/Mc1r), increasing antioxidant defense (ApoE), and suppressing inflammatory signaling (IL-17). This natural-biomimetic hybrid design leverages biocompatible materials to co-target multiple pathological axes, offering a novel self-adaptive approach for microenvironmental rehabilitation in vitiligo. Graphical Abstract

The CRISPR/Cas system is a powerful tool for molecular diagnostics, but its reliance on linear amplification constrains sensitivity, particularly for in situ imaging. Here, we discovered that phosphorothioate (PS)-modified activators can modulate Cas enzyme conformation via hydrophobic anchoring. By adjusting the PS modification sites, we achieved precise control over Cas activation and trans-cleavage resistance. Guided by this mechanism, we proposed a tailored design strategy featuring a “scattered” PS modification to engineer a linear “Coordinator” probe. This design effectively decouples Cas enzyme activation from substrate trans-cleavage resistance, enabling the construction of a Scattered PS Nucleic Acid-driven Cas Autocatalytic system (SACA). SACA achieves exponential amplification without external enzymes, enhancing Cas12a and Cas13a sensitivity by 50 000-fold and 10 000-fold, respectively. Furthermore, the superior biostability and structural simplicity of these linear probes endow SACA with excellent compatibility, facilitating precise in situ imaging of HPV16 and HPV18 mRNA in cervical cancer cells. This study not only advances the understanding of Cas enzyme regulation by chemically modified nucleic acids but also establishes a new paradigm for precise and efficient molecular diagnostics.

Multiple mutation detection is increasingly essential for clinical applications targeting the diagnosis, treatment decision-making, and prognosis assessment of thyroid cancer, especially for the limited amount of thyroid fine-needle aspiration (FNA) samples. However, there is a lack of cost-effective methods that can simultaneously achieve high sensitivity and high throughput for thyroid cancer. Herein, we present a novel multiplex mutation detection technology that integrates nucleotide enrichment (NE)-assisted specific identification of variant alleles with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), termed NE-MS, enabling the simultaneous identification of 26 somatic hotspot mutations in thyroid cancer. This method enhances sensitivity by removing the mass-modified dideoxynucleotide (ddNTP) matching the wild-type from the nucleotide mixture during the single-base extension reaction, leaving the mutant alleles available for analysis. NE-MS exhibits an 8-fold lower LOD compared to the regular MS method. This strategy provided an excellent diagnostic performance for thyroid cancer, revealing that multiple mutations are associated with poor prognosis in thyroid cancer patients. Accordingly, this study demonstrated that the NE-MS assay is a highly sensitive and reliable initial screening method for somatic mutation detection, as well as a diagnostic and prognostic tool for thyroid nodules.

Aptamer-based homogeneous immunoassays exhibit considerable potential in the domains of bioanalysis and biodiagnosis owing to their universality in analyzing both proteins and small molecules as well as their compatibility with nucleic acid amplification technologies. Nevertheless, the substantial signal leakage by nonspecific aptamer allostery poses a challenge to enhancing sensitivity further. Herein, we reported a T4 DNA polymerase-proofread DNA binding identifier (ReID). This strategy could harness the dual-enzymatic activity of T4 DNA polymerase to eliminate the leaked signal, thereby efficiently integrating target-induced aptamer allostery with subsequent polymerase chain reaction signal amplification. Moreover, we explored the regulation mechanism of dNTPs concentration on the dual-enzymatic activity of the T4 DNA polymerase. As a result, this strategy achieved an ultrasensitive protein detection limit of 8 fg/mL, validating the effectiveness of this proofreading approach. The universality was further confirmed by highly sensitive detection of small molecules. The exploration of ReID represents a significant advancement in the sensitivity and universality of immunoassays, even demonstrating the potential for multiple proteomic assays, offering a novel perspective for the development of high-performance homogeneous immunoassays.

RNA small fragment aberrances are associated with diseases by mediating a range of pathogenesis and pathological processes. DNA assembly‐based barcoding and amplification technologies are currently being actively explored for RNA in situ analysis. However, these modular integrated DNA assembly processes are inevitably accompanied with false positive signals caused by unexpected misassembly. Completely avoiding this phenomenon through simple and universal methods is challenging. Here, a novel dual‐input to dual‐output in situ analysis paradigm is proposed, aiming to improve target specificity through co‐recognition (dual‐input) and to eliminate false positive misassembly through fluorescent signal co‐localization (dual‐output). Based on this paradigm, Gemini molecular assembly co‐localization (GOAL) in situ imaging system is launched to accurately distinguish the fusion gene subtypes associated with chronic myeloid leukemia (CML), and to precisely report the proportion of minimum residual cancer cells in clinical samples by intelligent co‐localization counting and sorting. GOAL achieves highly sensitive and accurate genotyping recognition of 0.01% CML tumor cells and realizes fully automatic rapid diagnosis with a customized Intelligent Cell Image Sorter (iCis). iCis‐assisted GOAL represents an innovative and versatile molecular toolkit for accurate, rapid, user‐friendly, and professional‐independent profiling of cancer cells with RNA small fragment aberrances, providing efficient clinical decision support for disease diagnosis.