Diabetic cardiomyopathy (DCM), a severe complication of type 2 diabetes mellitus (T2DM), lacks specific and effective biomarkers for early diagnosis. This study constructed a plasma-specific spectral library by integrating proteomic and nonenzymatic glycation data from eight pretreatment workflows via data-dependent acquisition. Data-independent acquisition was then applied to profile plasma proteomes and glycation modifications in controls, DM patients, and DCM patients, revealing clear disparities in protein abundance and glycation modification patterns among the three groups. Functional enrichment analysis indicated that these differentially expressed proteins and modified peptides were involved primarily in immune responses, inflammatory processes, and metabolic pathways. Subsequently, parallel reaction monitoring was used to validate the proteins and glycation sites with significant changes. Specific peptides of complement 5 and specific glycation modifications on human serum albumin demonstrated a strong capacity to discriminate DCM from DM, achieving the highest area under the curve values of 0.97 in receiver operating characteristic analyses, underscoring their promising potential as DCM biomarkers. In conclusion, integrated proteomic and glycation modification analysis revealed candidate biomarkers for DCM diagnosis and offered novel insights into DCM pathogenesis.

Background:Asymptomatic hosts can shed pathogens without showing clinical symptoms, making them invisible to routine screenings and potent drivers of pathogen dissemination and epidemic outbreaks. The lack of reliable, cost-effective tools for large-scale identification of asymptomatic infections hampers early intervention and control strategies. Porcine deltacoronavirus (PDCoV), a zoonotic pathogen with potential for cross-species transmission, presents a critical case for improving such detection methodologies. Methods:We improved the IgG serodynamics-based epitope discovery method by integrating clustering and high-level analysis, which helped us identify linear epitopes with immunogenicity from a large number of candidate epitopes. Epitopes were filtered using negative sera identified by virus neutralization tests (VNT) to eliminate highly antigenic probes. These remaining low antigenicity probes were used to construct a protein-peptide hybrid microarray (PPHMPDCoV). The platform was applied to detect PDCoV-specific transiently produced IgGs (TPIs) in serum samples collected from pigs aged 28-174 days. Results:The PPHMPDCoV successfully detected asymptomatic PDCoV infections in pigs, particularly showing a peak infection rate of 15% at 45 days of age. The platform enabled not only the detection of asymptomatic carriers but also the characterization of infection stages. Conclusion:The study provides a novel, specific, and practical platform for detecting asymptomatic PDCoV infections based on serological TPI signatures. It offers early warning and disease prevention strategies in livestock and establishes a framework for future monitoring of potential interspecies transmission.

BACKGROUND:The prompt and precise diagnosis of active pulmonary tuberculosis (TB) is crucial for controlling this disease and yet it remains a global challenge. The objective of this study was to identify a set of microRNAs (miRNAs) whose expression in plasma could be used as a triage test for diagnosing TB. METHODS:A total of 879 plasma samples were collected in seven clinical centres from healthy individuals and patients displaying TB-like symptoms and/or radiological features consistent with TB. The samples were classified as TB, pneumonia, lung cancer and HC subgroups based on subsequent diagnostic assessments.We performed quantitative profiling of 264 plasma miRNAs in a training cohort (n=410) and an independent external test cohort (n=469). After dimensionality reduction and feature selection analysis, we identified nine discriminative miRNAs and used them to train an ensemble model in a training cohort using the scikit-learn library, which was subsequently evaluated in the external test cohort. RESULTS:The ensemble model showed notable accuracy in discriminating TB from non-TB patients, yielding areas under the curve (AUC) of 0.84 (95% CI 0.80 to 0.88) for the training cohort and 0.86 (95% CI 0.82 to 0.90) for the external test cohort. When tested against subgroups of laboratory confirmed and clinically diagnosed but unconfirmed TB, the AUC values were 0.89 (95% CI 0.85 to 0.93) and 0.83 (95% CI 0.79 to 0.88), respectively. In smear-negative confirmed TB patients, the AUC exceeded 0.83, with a sensitivity and specificity of 0.75. CONCLUSIONS:Our miRNA ensemble model, based on detecting a nine-miRNA expression biosignature in plasma, demonstrated promising ability to diagnose TB and distinguish it from other common lung diseases but further studies are needed to assess its clinical applicability. TRIAL REGISTRATION NUMBER:ChiCTR2000039734.

Viral myocarditis (VMC), caused by pathogens such as coxsackievirus B3 (CVB3), leads to severe cardiac injury and currently lacks specific therapeutic options. Here, we report a biomimetic antiviral strategy based on receptor engineering and intracellular gelation. By combining genetic and protein engineering, we generated a high-affinity Coxsackievirus and adenovirus receptor mutant (Mut-1_CAR) that markedly enhances the binding of host cardiomyocytes to CVB3. Using photochemical crosslinking, these engineered cells were converted into structurally stable, function-retaining gelated cells (PMs). PMs efficiently adsorb and neutralize virus particles, significantly reducing CVB3 plaque formation in vitro. In a murine model of viral myocarditis, PMs demonstrated excellent in vivo safety and biocompatibility while effectively lowering viral load and mitigating myocardial injury. This study establishes a “receptor enhancement + function fixation” approach for non-immune-dependent viral neutralization, providing a conceptual and technical foundation for the development of novel cell-based biomimetic antiviral therapies.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the benchmark platform for multiplex vitamin quantification, yet simultaneous determination of chemically disparate fat-soluble (FSVs) and water-soluble vitamins (WSVs) remains analytically challenging. We introduce a pragmatic dual-stream workflow that converts the intrinsic waste segments of any LC gradient into productive analytical time, delivering complete FSV and WSV profiles. Two 5-min chromatographic separations, optimized for fat-soluble and water-soluble vitamins respectively, are staggered through a six-port divert valve: while one column eluted its analytes to the mass spectrometer, the other was shunted to waste. This seamless overlap effectively doubles the throughput within 5 min. And every WSV elution window remained fully enclosed within the waste segment of the FSV gradient, with CVs < 10 % for the internal-standard peak areas across both vitamin classes. Moreover, method comparison between dual LC-MS/MS and single LC-MS/MS across twenty patient sera yielded Spearman's rho values ranging from 0.950 to 0.997, Passing-Bablok slopes spanning 0.884-1.128, and Bland-Altman biases below 8.3 %, confirming clinical concordance. Owing to its simple equipment requirements and setup process, this method should be highly accessible for other laboratories.

The ongoing arthropod-borne Chikungunya virus (CHIKV) highlights the requirements of rapid and accurate diagnostic methods to enhance the epidemic control. CRISPR diagnostic (CRISPR-Dx) technology holds promise, but the development of a highly efficient one-pot diagnostic system usually requires fine-tuning of the balance between isothermal amplification and Cas cleavage procedures. Here, we describe a simple method (psHOLMES) to create one-pot, two-step CRISPR-Dx systems, using photocleavable partially phosphorothioate-modified DNA (ppPS-DNA) to regulate Cas12a activity. Cas12a activity is first inactivated via binding of ppPS-DNA during the target sequence amplification procedure, which is then reactivated by ultraviolet (UV)-mediated photolysis of ppPS-DNA after amplification, triggering Cas12a trans-cleavage reactions. psHOLMES demonstrates attomolar sensitivity for CHIKV RNA detection and zero cross-reactivity against other related arboviruses. When applied to clinical samples, psHOLMES achieved 100% (50/50) accuracy and could detect CHIKV within 30 min. As traditional efforts for fine-tuning Cas cis-cleavage activity can be omitted, psHOLMES thus enables rapid development of one-pot CRISPR-Dx systems for clinical applications.

Antibodies are cornerstone molecules in immunity and biotechnology. Understanding the structural basis of specific antigen-antibody recognition is pivotal for informed antibody engineering and affinity optimization. However, the functional significance of binding-induced conformational changes, the principles governing binding interface organization, and the relationship between interface features and affinity remain incompletely understood. Here, we uncover the structural logic underlying antigen-antibody binding through integrated, multilevel analyses of large-scale structural datasets. Quantitative analyses reveal that binding-induced conformational rearrangements, although subtle, are functionally directed adjustments that refine interfacial complementarity. At the ensemble level, these adjustments significantly enhance electrostatic complementarity, whereas the optimization of geometric fit becomes evident upon stratification, revealing context-specific modes of refinement across different interface-area regimes. While the heavy chain typically dominates the binding, the total interface area is the more reliable correlate of affinity, with light and heavy chains contributing similarly by expanding the binding interface. Further analysis reveals the preferences of individual CDRs for specific interaction types. We also define six distinct interaction fingerprints of antigen-antibody binding, highlighting area-efficient and area-inefficient complexes with signature interaction profiles. Collectively, these findings delineate the mechanistic landscape of antigen-antibody binding and offer a roadmap for rational antibody engineering.

The development of robust immunoassays depends on high-affinity antibodies; however, rational design strategies remain challenging. Here, we present a structure-guided computational framework for antibody optimization and demonstrate its application in a clinical-grade chemiluminescence assay for C-reactive protein (CRP). Based on key topological residues identified from the cryo-EM structure of the CRP-HCAb complex, we rationally designed mutations within the complementarity-determining regions (CDRs). Molecular dynamics simulations predicted improved binding stability for the optimized variants, which was further validated by peptide-based binding assays. Upon recombinant expression, the engineered antibodies-HCAb3-CDR1-MT and HCAb4-CDR2-MT-exhibited enhanced specificity and affinity. When configured into a sandwich chemiluminescence immunoassay, the system showed excellent precision (CV < 5%) and strong resistance to interference from common serum components. In a clinical validation study involving 120 human serum samples, the assay demonstrated a strong correlation (R2 > 0.98) with a commercial method and outperformed it in repeatability when resolving discrepant samples. Collectively, this work establishes an integrated pipeline from atomic-level structure to functional clinical assay, offering a generalizable strategy for developing next-generation diagnostic antibodies.

Antigen-antibody specific recognition constitutes fundamental research in molecular drug design and immune diagnostics, where cumulative non-covalent interactions critically determine binding affinities. Current mechanistic understanding of antibody affinity optimization remains incomplete, hindering rational structure-based design of therapeutic antibodies and bispecific variants. This study presents four cryo-EM structures of C-reactive protein (CRP) complexed with heavy-chain antibodies (HCAbs) of varying affinities, resolved at 3.0-3.4 Å resolution. Comparative structural analysis reveals pronounced variations in binding modalities among affinity-differentiated HCAbs, while identifying critical determinants of engagement conformations, providing mechanistic insights for rational optimization of CRP-specific antibodies.

Aptamers, including nucleic acid and peptide aptamers, are small biological molecules whose development has consistently represented the forefront of science and technology. With advances in synthetic biology, bioinformatics, and cell biology, alongside the integration of multidisciplinary approaches, researchers have been able to construct aptamers of diverse structures and functions based on peptide self-assembly, thereby continuously driving innovation in this field. The maturation of various synthesis techniques has further facilitated the gradual translation of aptamers into the market. Supported by the establishment of aptamer information libraries, as well as their inherent excellent affinity and specificity, aptamers can now be synthesized, chemically modified, and applied across a broad spectrum of biomedical scenarios. They function not only as therapeutic agents and diagnostic probes, but also as biosensing tools and delivery vehicles for other drugs. These characteristics underscore the significance of aptamer development within the field of molecular recognition. In this paper, we conduct a comprehensive review of various research directions centered on their targeting properties, including their use as therapeutic and diagnostic agents, biosensors, platforms for new drug development, and drug delivery vehicles.

Infectious diseases pose a major challenge to public health worldwide. In recent years, vector-borne and zoonotic diseases have emerged as major public health threats. Effective prevention, control, and essential monitoring strategies are required to combat the rising global incidence and prevalence of infectious diseases. The frequency of infectious disease outbreaks has increased in the past several decades, and this trend has been predicted to be likely to continue. To effectively identify public health threats, and monitor and alert against infectious diseases, we obtained surveillance data from Shusi Tech’s Global Epidemic Information Monitoring System and conducted a comprehensive analysis of outbreak timing and location from the beginning of the year to December of 2024.

Arthropod-borne viruses (arboviruses) pose significant global health risks, yet their rapid detection remains challenging due to limitations in conventional diagnostics, including sensitivity and contamination risks. While CRISPR-based platforms have improved nucleic acid detection, current one-pot strategies still rely on target sequence-dependent suppression mechanisms, requiring laborious optimization for each target and limiting adaptability to emerging pathogens. Here, we report a universally applicable photocontrolled CRISPR/Cas12a system that eliminates target-specific optimization through engineered 3 '-modified crRNA. Mechanistic studies revealed that crRNA bearing 3 '-end modifications broadly inhibits Cas12a activity by inducing conformational distortion in the Cas12a/crRNA-DNA ternary complex. To temporally control this suppression, we engineered a photocleavable (PC) DNA linker between the crRNA 3 ' terminus and the additive, thereby decoupling amplification and detection phases in a single reaction. UV-triggered cleavage of the linker restores Cas12a activity without target sequence constraints, enabling a fully integrated one-pot system. This approach achieves 1 aM sensitivity, distinguishes yellow fever virus (YFV) from other arboviruses with excellent specificity, and demonstrates 100 % concordance with RT-qPCR in clinical samples. By integrating universal suppression regulation with precise photocontrol, our platform provides a contamination-free solution for point-of-care diagnostics, addressing critical needs in outbreak response and global health security.

Developing a preamplification-free and sensitive clustered regularly interspaced short palindromic repeats (CRISPR)-based method is significant but still extremely challenging for microRNA (miRNA) detection. Here we present a combination of a CRISPR/Cas13a-based reaction with a lateral flow biosensor, which enables the quantitative and colorimetric readout of preamplification-free miRNA detection at room temperature. In this work, the reaction principle and the structure of the lateral flow strip are well-designed to achieve surface-enhanced Raman scattering (SERS)/colorimetric dual-signal "turn-on" response of target miRNA. The CRISPR/Cas13a Reporter is engineered with a DNA-RNA splicing structure to generate DNA cleavage products and reduce nonspecific collateral cleavage. Without the need for nucleic acid preamplification strategy, the developed CRISPR/Cas13a-driven lateral flow biosensor enables the microRNA-21 (miR-21) detection at room temperature with a readout time of 10 min and a total process time of less than 45 min, achieving an impressive limit of detection of 8.96 aM by SERS and 1 fM by visualization, respectively. Moreover, the platform demonstrated excellent recovery rates in spiked human serum samples. The proposed CRISPR/Cas13a-driven, dual-signal "turn-on"-responded lateral flow platform has the potential to simultaneously meet the requirements of convenient point-of-care visualization detection and more accurate and sensitive SERS detection of miR-21, offering a cost-effective, rapid, and reliable tool for early cancer diagnosis.

In the first half of 2025, cholera, mpox, measles, dengue fever, chikungunya, malaria, and Zika virus spread globally, and severe outbreaks occurred in the Democratic Republic of the Congo (DRC) and Afghanistan. The primary drivers were enhanced pathogen adaptability and healthcare system vulnerabilities. These diseases showed remarkable spatial heterogeneity and dynamic evolution, influenced by climate, healthcare resources, and vector control. Temperature and precipitation played major roles in mosquito-borne disease transmission, and rising temperatures and altered precipitation patterns extended the zones of vector suitability. Four major challenges in arboviral disease control are summarized herein: vector ecology complexity, diagnostic-therapeutic access disparities, fragility of public health infrastructure, and climate-driven transmission expansion. Future prospects and recommendations include enhancing international cooperation, innovating prevention and control technologies, boosting public health system capacity, and addressing climate change to decrease the risk of mosquito-borne disease transmission.

Chemotherapy resistance presents a major challenge in the treatment of hepatocellular carcinoma (HCC), with the underlying molecular mechanisms largely unknown. This study aimed to investigate the role of tissue factor pathway inhibitor 2 (TFPI2) in modulating HCC chemosensitivity. We explored the impact of TFPI2 on sorafenib sensitivity in patient-derived organoids and mouse models using immunofluorescence analysis, chromatin immunoprecipitation, and RNA immunoprecipitation. We observed the downregulation of TFPI2 in HCC, and its deletion in mice (TFPI2HKO) accelerated DEN-induced liver tumorigenesis. Notably, TFPI2 overexpression increased sorafenib sensitivity in HCC organoids and in vivo models. Mechanistic insights indicated that TFPI2 stabilizes the mRNA of growth arrest and DNA damage-inducible alpha (GADD45A) by engaging the cell cycle and apoptosis regulator 2 (CCAR2), promoting GADD45A-mediated DNA damage and inhibiting homologous recombination repair. Furthermore, TFPI2 protects CCAR2 from ubiquitination-induced degradation by associating with the deubiquitinating enzyme BRCC3. We identified polydatin, a resveratrol glycoside, which upregulates TFPI2 and synergistically enhances the chemosensitizing effect of sorafenib in organoids and in vivo. TFPI2 plays a critical role in CCAR2-GADD45A-induced DNA damage repair, providing a strategy to enhance HCC chemosensitivity. Our findings elucidate the molecular intricacies of chemoresistance in HCC and reveal a potential therapeutic target for alleviating this resistance.

Purpose:This study aims to conduct a whole-genome analysis of the isolated strain HhutSZ1, providing more reliable clinical experience for the treatment of patients infected with such bacteria. Patients and Methods:A patient with IgA nephropathy failed to respond to treatment with ciprofloxacin for an infection. A rare strain of Herbaspirillum was isolated from the patient's blood. The VITEK MS Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) System and VITEK 2 Automated Microbial Identification System were used to conduct a preliminary identification and antibiotic susceptibility tests. Then we performed whole-genome sequencing of this strain along with comparative genomic analysis. Results:VITEK 2 System identified HhutSZ1 as Burkholderia cepacia, while the VITEK MS System identified it as Herbaspirillum huttiense with a high confidence coefficient. The phylogenetic tree based on the 16S rRNA gene showed that HhutSZ1 belonged to the genus Herbaspirillum. Average Nucleotide Identity (ANI) analysis showed that the scores of this strain compared with other Herbaspirillum huttiense strains were all lower than 95, confirming that this strain did not belong to Herbaspirillum huttiense. The patient's infection failed to resolve despite ciprofloxacin treatment. Subsequent antimicrobial susceptibility testing revealed that HhutSZ1 exhibited intermediate resistance to ciprofloxacin, which is consistent with the clinical treatment failure. Six genes were detected in the Comprehensive Antibiotic Resistance Database (CARD). And among them, five RND family efflux pump genes resistant to fluoroquinolone were all located on chromosomes. Conclusion:For the genus Herbaspirillum, mass spectrometry identification cannot accurately identify the species. The analysis based on 16s rRNA combined with ANI can be more accurate. Some Herbaspirillum spp may have inherent resistance to fluoroquinolone antibiotics. In conclusion, our findings suggest that the low detection rate and low drug resistance of this strain cannot be overlooked, and the study provides valuable clinical insight for managing infections in immunocompromised patients.

Sepsis remains the leading cause of in-hospital mortality in critically ill patients. Platelet distribution width (PDW), an indicator of platelet activation and variability, is associated with inflammation and coagulation dysfunction during sepsis. However, dynamic changes in PDW and their association with patient outcomes remain unexplored. This study investigated the relationship between changing PDW trends and in-hospital mortality in critically ill patients with sepsis using machine learning techniques for robust analysis. In the model development cohort, inpatient admissions fulfilling the sepsis 3.0 criteria in the Intensive Care Unit of Shenzhen People’s Hospital were analyzed. PDW measurements were obtained at six-time points: First Day (D1), Second Day (D2), Third Day (D3), and the last three days before discharge (LD-3, LD-2, and LD-1). PDW was compared between survivors and non-survivors. Group-based trajectory modeling identified distinct PDW trajectory groups, and patient characteristics and outcomes were analyzed. The model was externally validated at a second hospital using identical inclusion criteria. A total of 1,090 and 429 patients with sepsis were included in the development and validation cohorts, respectively. Four distinct PDW trajectory groups emerged in the development cohort: “PDW Rapidly Increasing Group” (n = 174; 15.96

We investigated genomic evolution of vancomycin-resistant Enterococcus faecium (VREF) during an outbreak in Shenzhen, China. Whole-genome sequencing revealed 2 sequence type 80 VREF subpopulations diverging through insertion sequence-mediated recombination. One subpopulation acquired more antimicrobial resistance and carbohydrate metabolism genes. Persistent VREF transmission underscores the need for genomic surveillance to curb spread.

With the development of e-commerce, noninvasive mail inspection is becoming particularly prominent. Terahertz waves have fingerprint spectrum characteristics and can penetrate nonpolar materials. Terahertz waves are ideal for the nondestructive identification of harmful substances hidden in the mail. However, the gaps between mail packages and samples affect the accuracy of the inspection. In this study, the influence of irregular gaps was analyzed using a model sample under envelope occlusion. A spectral reconstruction method based on Voigt and asymmetric least squares (AsLS) fitting is proposed. Principal component analysis (PCA) results showed that the reconstructed spectral data were easier to identify and the root mean square error (RMSE) of quantitative analysis was the smallest. PCA–support vector machine (SVM) and convolutional neural network (CNN) classification models were used to verify the effectiveness of this method.

C-Reactive protein (CRP) is a key biomarker for evaluating inflammation levels and estimating cardiovascular risk. However, current CRP detection methods rely on monoclonal antibodies (mAb), which possess shortcomings such as a lengthy preparation cycle, high cost, and poor repeatability. To address these challenges, we explored the potential of peptide aptamers as an alternative to mAb for CRP detection. Using some bioinformatics approaches, we designed and optimized peptide aptamers, selecting the dominant peptide aptamer C9m (KWRWRFRLSR) through experimental validation for its specific recognition of CRP. We then established a sandwich ELISA detection system combining C9m with CRP mAb. This system demonstrated a detection limit of 22.275 ng/mL CRP and exhibited excellent specificity, with no cross-reactivity observed with human serum albumin or γ-globulin. The method also showed high reproducibility, with intra- and inter-assay coefficients of variation (CV) less than 15 %, meeting laboratory testing standards. Furthermore, comparison with clinically used immunoturbidimetry revealed high consistency (r = 0.9891).