Amplification-free detection of genetically modified crops via spatial confinement CHA/HCR-aided surface-enhanced resonance Raman spectroscopy.

To distinguish genetically modified (GM) crops, we developed a target RNA amplification-free surface-enhanced resonance Raman scattering (SERRS) biosensor for the CaMV35S promoter RNA in GM crops by integrating enzyme-free, spatial confinement catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR). A magnetic-plasmonic composite probe modified with the first sequence of CHA was first fabricated. A small amount of target RNA can activate it to initiate the CHA, and the CHA efficacy benefits from the spatial confinement effect provided by the composite probe.

IL-2 and IFN-γ Secretion of Activated Jurkat T Cells a Microdroplet-SERS based Single-Cell Immunoassay (Drop-SCIA).

Studying the characteristics of T cell activation and cytokine secretion is crucial for understanding the cell-mediated immunological response (CMI). To assess this purpose, we present a droplet-based single-cell immunoassay platform, named Drop-SCIA, which uniquely integrates surface-enhanced Raman spectroscopy (SERS) with homogeneous-phase immunoassay, enabling highly sensitive, multiplexed cytokine detection at the single-cell level and offering superior target enrichment efficiency compared to the widely used interface-based ELISpot assay. Using this platform, we analyzed Jurkat T cell activation and further profiled IL-2 and IFN-γ secretion following coculture with normal breast epithelial cells (MCF-10A) and breast cancer subtypes (MCF-7, MDA-MB-231).

CRISPR/Cas-Based Electrochemical Biosensor for Human Immunodeficiency Virus-1 Nucleic Acid Amplification-Free Detection to the Attomolar Level.

Aiming at realizing field detection during HIV-1 patient screening in developing countries and point of care testing (POCT) of virus levels in HIV-1 patients receiving medication, an urgent demand for portable nucleic acid detection technology with low cost and sensitivity is raised. To solve this, a CRISPR/Cas13a-based electrochemical detection platform by a multiple combined crRNAs strategy is developed. This sensing platform is based on the Ion Current Rectification regulation through a Porous Anodic Alumina membrane decorated with ssRNA chains, which are trans-cleavage substrates for activated Cas13a and become shorter when the target gene exists, resulting in an altered motion of transmembrane ions.

Subchronic exposure to CeCl promotes macrophage ferroptosis and atherosclerotic plaque formation by inhibiting the Keap1/Nrf2/GPX4 pathway.

Cerium chloride (CeCl), a prevalent by-product of rare earth mining, accumulates in the biota and environment of mining regions, including plants, animals, water resources, and air, posing potential health risks to local residents. Atherosclerosis (AS) is the pathological basis of cardiovascular disease. However, the relationship between subchronic CeCl exposure and AS progression, along with the underlying regulatory mechanisms by which CeCl modulates AS, remains unclear.

Molecule-Responsive SERS Sensors for Urine Diagnosis of Kidney Diseases Enhanced by Neural Networks.

Early diagnosis of kidney disease is crucial for treatment and prognosis. Compared with kidney biopsy, a noninvasive urine-based diagnosis method of kidney disease can be more convenient and less painful for patients. Urine is closely associated with kidney disease, including nephritis, kidney failure, and kidney cancer.

Exploring Lipid Droplets for Enhancing Cytoskeletal Stability and Repair Capacity via Carbonized Polymer Dot-Assisted Fluorescence Imaging.

Lipid droplets (LDs), as dynamic organelles, are regarded as core carriers of energy storage and participate in cell signaling and metabolic process regulation. The interaction between LDs and cytoskeletons may profoundly affect cell growth and pathological processes; however, direct evidence is absent. Our study employed an LD-targeted carbonized polymer dot (CPD) that exhibits high photostability and superior imaging capabilities with confocal fluorescence imaging to visually observe the behaviors of LDs in reinforcing cytoskeletons and assisting cytoskeletal repair.

Surface-Enhanced Raman Spectroscopy for Biomedical Applications: Recent Advances and Future Challenges.

The year 2024 marks the 50th anniversary of the discovery of surface-enhanced Raman spectroscopy (SERS). Over recent years, SERS has experienced rapid development and became a critical tool in biomedicine with its unparalleled sensitivity and molecular specificity. This review summarizes the advancements and challenges in SERS substrates, nanotags, instrumentation, and spectral analysis for biomedical applications.

Surface-enhanced Raman spectroscopy (SERS)-based urine biopsy for kidney transplant patients.

For kidney transplantation patients, monitoring potential complications such as immune rejection and infection has always been a clinical challenge since the existing detection technology faces difficulties due to limited information, insufficient sensitivity and specificity, and lack of continuous dynamic monitoring ability. Herein, we employed surface-enhanced Raman spectroscopy (SERS) for liquid biopsy, which has non-invasive and highly sensitive characteristics, to assess the physiological status of kidney transplant patients. Two SERS methods were applied to analyze the changes in urine at different stages of renal transplantation.

Three-Dimensional SERS-Active Hydrogel Microbeads Enable Highly Sensitive Homogeneous Phase Detection of Alkaline Phosphatase in Biosystems.

Alkaline phosphatase (ALP) is a biomarker for many diseases, and monitoring its activity level is important for disease diagnosis and treatment. In this study, we used the microdroplet technology combined with an laser-induced polymerization method to prepare the Ag nanoparticle (AgNP) doped hydrogel microbeads (HMBs) with adjustable pore sizes that allow small molecules to enter while blocking large molecules. The AgNPs embedded in the hydrogel microspheres can provide SERS activity, improving the SERS signal of small molecules that diffuse to the AgNPs.

Revealing mitochondrial microenvironmental changes triggered by ferroptosis regulation.

Ferroptosis is a unique form of iron-dependent cell death characterized by dramatic ultrastructural changes in mitochondria. Since mitochondria are intracellular energy factories and binding sites for producing reactive oxygen species, there is increasing evidence that mitochondria are closely related to ferroptosis and play a crucial role in the regulation and execution of ferroptosis. The pH of the mitochondrial microenvironment is an important parameter for cellular physiological activities.

Cervical cancer diagnosis model using spontaneous Raman and Coherent anti-Stokes Raman spectroscopy with artificial intelligence.

Cervical cancer is the fourth most common cancer worldwide. Histopathology, which is currently considered the gold standard for cervical cancer diagnosis, can be time-consuming and subjective. Therefore, there is an urgent need for a rapid, objective, and non-destructive cervical cancer detection technique.

Super-Resolution Microscopic Imaging of Lipid Droplets in Living Cells via Carbonized Polymer Dot-Based Polarity-Responsive Nanoprobe.

Lipid droplets (LDs) are dynamic subcellular organelles that participate in various physiological processes, and their abnormality can also lead to various diseases. Tracing the dynamics of LDs in living cells will be valuable for understanding cell physiological states. Here, we employed a structured light illumination super-resolution imaging assisted with a carbonized polymer dot (CPD)-based fluorescence nanoprobe to track the travel paths of LDs and other organelles.

Identification of Molecular Profile of Cell Membrane via Magnetic Plasmonic Nanoprobe.

Cell membranes are primarily composed of lipids, membrane proteins, and carbohydrates, and the related studies of membrane components and structures at different stages of disease development, especially membrane proteins, are of great significance. Here, we investigate the chemical signature profiles of cell membranes as biomarkers for cancer cells via label-free surface-enhanced Raman scattering (SERS). A magnetic plasmonic nanoprobe was proposed by decorating magnetic beads with silver nanoparticles, allowing self-driven cell membrane-targeted accumulation within 5 min.

Comparison of a Supervised Home-Based Tele-Rehabilitation with Center-Based Pulmonary Rehabilitation: Protocol for a Randomized Non-Inferiority Multicenter Study in Ningxia.

Background: Pulmonary rehabilitation (PR) is an effective intervention for people with chronic obstructive pulmonary disease (COPD). However, fewer than 5% of eligible individuals receive pulmonary rehabilitation, largely due to limited by the accessibility of rehabilitation and difficulties associated with travel and transport. Supervised home-based tele-rehabilitation (SHTR) is an alternative model to center-based pulmonary rehabilitation.

In-droplet multiplex immunoassays for hypoxia-induced single-cell cytokines.

Cytokine expression is an important biomarker in understanding hypoxia microenvironments in tumor growth and metastasis. In-droplet-based immunoassays performed above the target cell membrane were employed to track the cytokines of single cells with the aid of three types of immuno-nanoprobes (one capture nanoprobe and two reporter nanoprobes). Single cells and nanoprobes were co-packaged in water-in-oil microdroplets (about 100 μm in diameter) using a cross-shaped microfluidic chip.

An atomic force microscopy and total internal reflection fluorescence microscopy correlated system (AFM-TIRF) for fluorescence imaging and spectroscopy of a single particle.

Combining atomic force microscopy (AFM) with other optical microscopic techniques is pivotal in nanoscale investigations, particularly leveraging the surface-sensitive properties of total internal reflection fluorescence microscopy (TIRF). A novel design that integrates AFM with a multi-wavelength TIRF is displayed, providing simultaneous fluorescence imaging and spectral acquisition capabilities. We elaborate on the considerations in the instrument design process and demonstrate the performance and potential applications of the instrument through fluorescence imaging and spectroscopy testing of individual nanoparticles.

A review of clinical use of surface-enhanced Raman scattering-based biosensing for glioma.

Glioma is the most common malignant tumor of the nervous system in recent centuries, and the incidence rate of glioma is increasing year by year. Its invasive growth and malignant biological behaviors make it one of the most challenging malignant tumors. Maximizing the resection range (EOR) while minimizing the impact on normal brain tissue is crucial for patient prognosis.

Fabrication of High-Negatively Charged Bicelle-Mediated Supported Lipid Bilayer.

Supported lipid bilayers (SLBs), two-dimensional lipid films formed on a solid-supporting substrate, serve as models for biomembranes and exhibit remarkable potential in chemistry, biology, and medicine. However, preparing SLBs with highly negatively charged contents on the negatively charged surface by overcoming electrostatic repulsion remains a challenge. Here, a creative bicelle-mediated and divalent cation-free SLB preparation method with the assistance of phosphate-buffered saline (PBS) solution was proposed, which can form the SLBs containing 50% DOPS or 30% CL on the silica surface monitored by a quartz crystal microbalance with dissipation (QCM-D).

Engineering of 4-hydroxyphenylacetate 3-hydroxylase derived from Pseudomonas aeruginosa for the ortho-hydroxylation of ferulic acid.

Polyphenolic compounds have natural antioxidant properties, and their antioxidant activity is usually related to the number and position of hydroxyls. Here, we successfully applied the engineered 4-hydroxyphenylacetate 3-hydroxylases (4HPA3Hs) derived from Pseudomonas aeruginosa to catalyze ferulic acid (FA) synthesis of ortho-hydroxyferulic acid (5-hydroxyferulic acid, 5-OHFA). Through optimization of co-expression, the oxygenase component (PaHpaB) and the reductase component (PaHpaC) in E.

Policies for recovery from drug use: Differences between public stigma and perceived stigma and associated factors.

Introduction: Public stigma towards people who use drugs is widespread and places obstacles in way of their recovery. Previous studies have used different approaches to measure public stigma, resulting in a notable gap in the understanding of the relationship between it and its associated factors. Some studies measure public stigma by assessing stigma perceived by those who use drugs, while others investigate attitudes towards them among the general public.

4-Hydroxyphenylacetate 3-Hydroxylase (4HPA3H): A Vigorous Monooxygenase for Versatile -Hydroxylation Applications in the Biosynthesis of Phenolic Derivatives.

4-Hydroxyphenylacetate 3-hydroxylase (4HPA3H) is a long-known class of two-component flavin-dependent monooxygenases from bacteria, including an oxygenase component (EC 1.14.14.

Sensitive detection of genetically modified maize based on a CRISPR/Cas12a system.

With the vigorous development of biotechnology, genetically modified organisms (GMOs) have become more and more common. In order to effectively supervise and administrate them, the rapid and accurate detection of GMOs is urgently demanded. Here, GMO gene-specific sensing methods based on colorimetry and surface-enhanced Raman scattering (SERS) were proposed based on the lateral branch cleavage function of the CRISPR/Cas12a system.

Biosynthesis of 3-Hydroxyphloretin Using Rational Design of 4-Hydroxyphenylacetate 3-Monooxygenase.

The compound 3-hydroxyphloretin is a typical dihydrochalcone that can be obtained in plants by the 3-hydroxylation of phloretin. Here, the flavin-dependent two-component monooxygenase (HpaBC) derived from was used to convert phloretin into 3-hydroxyphloretin. Following molecular docking and sequence alignment, modifications to the substrate pocket and loop of HpaBC were rationally designed, and mutant residues were selected.