Recent, outbreaks of new emergency zoonotic diseases have prompted an urgent need to develop fast, accurate, and portable screening assays for pathogen infections. Recombinase polymerase amplification (RPA) is sensitive and specific and can be conducted at a constant low temperature with a short response time, making it especially suitable for on-site screening and making it a powerful tool for preventing or controlling the spread of zoonoses. This review summarizes the design principles of RPA-based biosensors as well as various signal output or readout technologies involved in fluorescence detection, lateral flow assays, enzymatic catalytic reactions, spectroscopic techniques, electrochemical techniques, chemiluminescence, nanopore sequencing technologies, microfluidic digital RPA, and clustered regularly interspaced short palindromic repeats/CRISPR-associated systems. The current status and prospects of the application of RPA-based biosensors in zoonoses screening are highlighted. RPA-based biosensors demonstrate the advantages of rapid response, easy-to-read result output, and easy implementation for on-site detection, enabling development toward greater portability, automation, and miniaturization. Although there are still problems such as high cost with unstable signal output, RPA-based biosensors are increasingly becoming one of the most important means of on-site pathogen screening in complex samples involving environmental, water, food, animal, and human samples for controlling the spread of zoonotic diseases.

The number of words: 4645, the number of figures: 4, the number of tables: 1The outbreak of COVID-19 in December 2019 caused a global pandemic of acute respiratory disease, and with the increasing virulence of mutant strains and the number of confirmed cases, this has resulted in a tremendous threat to global public health. Therefore, an accurate diagnosis of COVID-19 is urgently needed for rapid control of SARS-CoV-2 transmission. As a new molecular biology technology, loop-mediated isothermal amplification (LAMP) has the advantages of convenient operation, speed, low cost and high sensitivity and specificity. In the past two years, rampant COVID-19 and the continuous variation in the virus strains have demanded higher requirements for the rapid detection of pathogens. Compared with conventional RT–PCR and real-time RT–PCR methods, genotyping RT-LAMP method and LAMP plus peptide nucleic acid (PNA) probe detection methods have been developed to correctly identified SARS-CoV-2 variants, which is also why LAMP technology has attracted much attention. LAMP detection technology combined with lateral flow assay, microfluidic technology and other sensing technologies can effectively enhance signals by nucleic acid amplification and help to give the resulting output in a faster, more convenient and user-friendly way. At present, LAMP plays an important role in the detection of SARS-CoV-2.

The western part of north-eastern China is mostly covered by soda soil, which has a significant impact on the growth, breeding and production of crops. The present study investigates the physiological responses and metabolic adaptation of Chlamydomonas reinhardtii P.A.Dangeard (C. reinhardtii) under different types of alkaline stress and the potential application to soda soil improvement and plants in response to alkaline stress. The soda–saline soil was treated with C. reinhardtii for 5 and 120 days, respectively, and the chemical properties of the soil were determined. The microalga was cultured for a period of 12 days, with and without an alkaline medium, and then the metabolic profiles of the microalgae were then analyzed by metabolomics. Results showed that C. reinhardtii exhibited a high level of tolerance to 40 mM NaHCO₃, pH 8.3, and pH 10.3 with an increase in incubation time. Moreover, C. reinhardtii has been demonstrated to ameliorate selected physicochemical properties of soda–saline soil by reducing soil pH, electrical conductivity (EC), and the contents of CO₃²⁻ and HCO₃⁻ by 40–100% (w/w, wet biomass) addition for 120 cultivation days, respectively. It was demonstrated that rice seedlings were less affected by alkaline stress in the presence of C. reinhardtii. Furthermore, C. reinhardtii has the capacity to adapt to alkaline environments by regulating key pathways such as cofactor biosynthesis, glycerophospholipid metabolism, amino acid biosynthesis, purine metabolism, carbon metabolism, tropane, piperidine, and pyridine alkaloid biosynthesis. This adaptation is characterized by the accumulation of metabolites, including saccharides, amino acids and organic acids. These interact with high pH, CO₃²⁻ and HCO₃⁻ ions. This study elucidates that C. reinhardtii has the capacity to improve soda–saline soil and respond well to diverse alkaline stresses, attributable to its intricate metabolic profile.

Background Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited targeted therapeutic options and marked heterogeneity in response to immune checkpoint blockade. Shikonin, a natural naphthoquinone compound, has shown antitumor and immunomodulatory activities, but its potential molecular targets and immune-related transcriptional programs in TNBC remain incompletely characterized. Methods We integrated systems pharmacology, TCGA-BRCA bulk transcriptomic data, molecular docking, immune infiltration analysis, prognostic modeling, and single-cell RNA sequencing data from GSE176078. Putative shikonin-associated TNBC targets were identified by intersecting predicted shikonin targets, TNBC-related genes, and differentially expressed genes. Functional enrichment analysis was performed to characterize biological pathways. Candidate immune-related genes were prioritized according to their correlation with CD274 expression. The resulting signature was evaluated for associations with immune checkpoint genes, immune cell infiltration, immune activation, computationally inferred immunotherapy-related phenotypes, and overall survival. Single-cell analysis was used to define the cellular distribution of signature genes within the TNBC tumor microenvironment. Results Seventy candidate shikonin-associated TNBC targets were identified and were enriched in immune-related and oncogenic pathways, including PD-1/PD-L1 checkpoint signaling, chemokine signaling, PI3K-Akt signaling, and MAPK signaling. A CD274-prioritized 10-gene immune signature comprising ITK, CCR4, TAP1, JAK3, PIM2, LYZ, SYK, HCK, SOAT1, and PLA2G7 was identified by integrating positive CD274 correlations, false-discovery-rate control, and established immune relevance. High signature scores were associated with elevated immune checkpoint expression, increased immune infiltration, a stronger immune activation index, and lower predicted immune evasion. Molecular docking provided computational support for potential binding compatibility between shikonin and the encoded proteins. At single-cell resolution, the signature genes were distributed across lymphoid, myeloid, epithelial, and stromal compartments, indicating a multicellular immune-related niche. Conclusion This study identifies a shikonin-associated immune signature that reflects an inflamed and checkpoint-enriched tumor microenvironment in TNBC. These findings provide candidate molecular features and a hypothesis-generating framework for further experimental evaluation of the potential immunomodulatory relevance of shikonin in TNBC.