Ultrathin layer chromatography is an efficient method that is fast and requires a small amount of sample for the separation. This method may be valuable in for the separation of biological samples in many different industries such as pharmaceutical analyses as well as clinical analyses. This is the first example of the use of nanofibers of DEAE-cellulose for UTLC for protein separations. This is combined with the detection of the proteins using paper spray ionization mass spectrometry.

Cardiolipins (CL) are a mitochondria-specific family of phospholipids that play central roles in mitochondrial function. Imbalance in CL metabolism, especially excessive CL oxidation, leads to mitochondrial dysfunction, apoptosis, and inflammation, contributing to age-related diseases. As of yet no comprehensive methods have been developed to assess CL, oxidized CL (oxCL), and monolyso-CL (MLCL) species.

Researchers are developing customized capillary electrophoresis (CE) instruments due to their affordability, and versatility. CE's simplicity, compared to chromatography-based methods, allows for low-cost designs using commercially available components. Injection methods, however, necessitate particular attention because of the high reproducibility required for quantitative determinations. Traditional manual hydrodynamic and electrokinetic injections involve one or more actions by the user thus increasing the number of potential sources of error. Automation efforts, such as using syringe pumps and pressure controllers, have improved injection precision, achieving results comparable to commercial systems. Despite progress, many methods still require manual sample loading and flushing, highlighting the need for further innovation in CE automation.

Simultaneous inhibition of α-glucosidase (α-Glu) and protein tyrosine phosphatase 1B (PTP1B) offers a promising therapeutic strategy for type 2 diabetes (T2D). Despite the natural product potential in T2D management, existing screening methods predominantly target single enzymes, whereas multi-enzyme approaches are often costly and less practical. A simple, cost-effective method for identifying multi-target inhibitors is therefore urgently needed.

Cardiac troponin I (cTnI) is a crucial diagnosis biomarker for acute myocardial infarction (AMI). Early accurate determination of the concentration of cTnI significantly decreases the death rate of AMI. Compared with classic methods, dual-mode sensors for cTnI determination could help reduce the false positive rate.

Aquatic viruses cause devastating diseases in aquaculture, severely limiting production and resulting in significant economic losses. The giant salamander iridovirus (GSIV), a member of the genus Ranavirus, is the only virus reported to infect giant salamanders, posing a severe threat to the farming industry. Currently, there are no effective strategies available for its control. In this context, a reliable diagnostic tool for the rapid detection of GSIV is crucial to mitigate its impact. This study developed a nanobody-based immunoassay for the rapid and reliable detection of GSIV. GSIV was cultured and used to immunize an alpaca, A phage library with an original diversity of 1.89 × 10 PFU was established, and six nanobodies were identified after three rounds of panning. Among them, HC-2 exhibited superior performance and was used to develop a highly sensitive ELISA method employing streptavidin-PolyHRP (SA-PolyHRP) as a signal amplification strategy. The assay achieved a detection limit of 3.3 × 10 PFU/mL and demonstrated high specificity without cross-reactivity. Practical application was validated in infected giant salamander samples, underscoring its diagnostic potential. This work provides a robust tool for GSIV diagnosis and showcases the potential of nanobodies in advancing aquaculture diagnostics.

Peroxynitrite (ONOO) and lipid droplets (LDs) are crucial substances essential for maintaining normal physiological functions in biological systems. They play pivotal roles as biomarkers in the initiation and progression of various diseases, such as epilepsy. Therefore, the simultaneous detection of ONOO and LDs in epilepsy disorders is of great importance. Here, we discovered that the fluorescence probe composed of trifluoromesulfonate and fluorophore can not only be used as the recognition site of ONOO, but also has the property of LDs targeting. Therefore, we reasonable designed and synthesized a dual-channel fluorescent probe CBT, capable of simultaneously monitoring ONOO and LDs. CBT exhibited exceptional dual-response properties: firstly, upon specific reaction with ONOO, the resulting product BHD emitted a robust red fluorescent signal in the near-infrared region (749 nm); secondly, CBT selectively targeted and labeled LDs, emitting green fluorescence at 482 nm for effective LDs tracking. The signals from these two detection channels did not overlap, which significantly enhanced the accuracy and reliability of detection. Based on these characteristics, CBT has been successfully utilized in real-time imaging of ONOO and LDs in epilepsy models of cells induced by various drugs. Notably, in a pentylenetetrazole (PTZ)-induced chronic epileptic mice model, CBT exhibited excellent efficacy in ONOO imaging, further confirming its considerable potential for practical applications. In summary, this study validated CBT as an efficient tool capable of simultaneous detection and differentiation of ONOO and LDs, presenting a novel and promising strategy for the early diagnosis and treatment of diseases such as epilepsy.

Real-time monitoring of hemolysis dynamics is essential for clinical diagnosis, ensuring transfusion safety, and supporting medical device development. Traditional methods such as spectrophotometry have limitations in real-time monitoring capabilities, often posing higher operational costs and restricted temporal resolution.

Advances in bio-analyte detection demonstrate the need for innovation to overcome the limitations of traditional methods. Emerging viruses evolve into variants, driving the need for fast screening to minimize the time required for positive detection and establish standardized detection. In this study, a SERS-active substrate with Au NPs on a regularly arranged ZrO nanoporous structure was utilized to obtain the SERS spectrum of inpatient samples from COVID-19 patients. Two analytical approaches were applied to classify clinical samples - empirical method to identify peak assignments corresponding to the target SARS-CoV-2 BA.2 variant, and machine learning (ML) method to build classifier models.

As a non-invasive liquid biopsy technology, the detection of circulating tumor cells (CTCs) overcomes the limitations of traditional tissue biopsy methods, enabling continuous sample collection and long-term dynamic monitoring. However, current CTCs analysis methods typically rely on cell size to separate and identify tumor cells, which fails to effectively distinguish tumor cells from different sources. In addition, existing methods are often constrained by limited antibody species, typically detecting only 2-3 molecular phenotypes. This narrow detection scope does not fully capture the heterogeneity of CTCs at the single-cell level, thus limiting its utility in precision diagnosis and personalized treatment. To address these challenges, it is urgent to develop CTCs detection methods that can simultaneously integrate comprehensive target and cell morphology information.

Traditional temperature-dependent models face difficulties in compact systems due to complex temperature control. This study introduces an electric field strength and runtime driven (E-t) band model to improve GE performance by correlating band behavior with electric field and runtime rather than temperature.

Triglycerides (TGs) play a crucial role in various physiological processes through the breakdown of their fatty acyl (FA) side chains. It has been demonstrated that not only the total levels of TGs but also the specific composition of FA side chains are vital for biological functions. However, biomedical studies that comprehensively identify FA compositions remain very limited. One of the reasons is the structural heterogeneity of TGs, with variability in their three fatty acyl chains posing significant challenges for TG analysis.

The coagulation process is inherently complex, and conventional diagnostic techniques are typically only capable of detecting single coagulation factors, lacking capacity to assess the overall coagulation function of the patient. Therefore, it is thus imperative to develop a novel, global coagulation assay for the diagnosis of hemorrhagic and thrombotic disorders and to evaluate the therapeutic efficacy of anticoagulant drugs. The ordered porous layer interferometry technology, developed using silica colloidal crystal (SCC) film as sensing substrate, is characterized by low cost, real-time, and convenient operation. Furthermore, entire biological events process can be detected by migration of the SCC film interference spectra.

Sulfatides are a class of sphingolipids which are abundant in the myelin sheet and oligodendrocytes, therefore they play a crucial role in the nervous system. Abnormal sulfatide excretion has been linked to several neurodegenerative disorders including metachromatic leukodystrophy (MLD) and multiple sulfatase deficiency (MSD). In MLD and MSD, sulfatide catabolism is impaired due to the reduced lysosomal arylsulfatase A (ARSA) activity resulting in an accumulation of sulfatides, which can be useful in a diagnostic setting. The current study aims to develop a method for semi-quantitation of urine sulfatides as a diagnostic tool for MLD and MSD.

Lipidated anorexigenic peptides are highly promising compounds for the treatment of obesity and related diseases. However, their exact mechanism of action still remains unknown. We labelled a lipidated analogue of an anorexigenic prolactin-releasing peptide (palm-PrRP31) with an extremely stable ClickZip lanthanide tag, facilitating tracking of the peptide within the organism. We then employed a separation method based on liquid chromatography combined with inductively coupled plasma mass spectrometry (LC-ICP-MS). This technique involved the use of an unconventional mobile phase containing 5 % 1,2-hexanediol in HO (v/v) with the addition of 2 % formic acid. Using a rapid. 6-min analysis, we were able to quantify the ClickZip tag - and thus indirectly the fate of the labelled peptides in the living organism - independently of free Ln ions. The detection limits for the various lanthanide chemical forms were extremely low, ranging between 0.9 and 3.4 ng/L. We demonstrated the suitability of the method for analysing real biological samples like blood plasma, and confirmed the accuracy of our results. Prior to LC-ICP-MS analysis, we optimised a process involving the microwave-assisted digestion of liver samples to preserve the integrity of the ClickZip tag. We also identified several metabolites of the labelled peptides in the liver, urine, and blood plasma, highlighting the utility of the method for revealing the mechanism of action behind the labelled lipopeptides.

Salmonella is a prevalent zoonotic pathogen that threatens food safety and human health. Owing to the large number of Salmonella species and their significant variations in pathogenicity and virulence, it is difficult to classify Salmonella strains quickly, which makes rapid detection of Salmonella outbreaks and research on foodborne diseases difficult.

Current available detection methods can not afford the direct and precise detection of trace arsenite (As(III)) in high-salinity water bodies. Therefore, the development of device with low limit of detection (LOD) for the early detection of As(III) in high-salinity water samples is of vital importance to secure environment and food safety.

Over the recent years, the investigations on wound dressings have been undergoing significant evolution, and now smart dressings with the function of the real-time monitoring of the wound states have been recognized as one of the most advanced treatment modalities. Among a variety of wound-related biomarkers, pH represents a promising candidate for in situ supervising the wound healing status. In this regard, a variety of optically pH sensing agents have been widely incorporated into different types of wound dressings.

Acetylcholinesterase (AChE) plays a critical role in maintaining nervous system homeostasis and coordinating essential biological reactions. AChE is an important biomarker for early diagnosis and treatment, including Parkinson's disease (PD), Alzheimer's disease (AD), and Huntington's disease (HD). Therefore, developing efficient and immediate sensing platforms for AChE detection is crucial for advancing early diagnostic tools. In this study, we developed a dual-mode colorimetric/photothermal sensing platform based on iodide ion-synergized covalent porphyrin-triazine backbone nanozymes (Zn-CTF/I) to detect AChE with high sensitivity and reliability. The synergistic interaction between iodide ions and zinc atoms effectively modulated the electronic structure of the catalytic active site, significantly enhancing the peroxidase-like (POD-like) activity of Zn-CTF/I. This enhancement led to a 10-fold reduction in the AChE detection limit compared to controls, with a minimum detection limit of 0.003 U L, outperforming other reported assays. The integration of temperature-based photothermal signals with colorimetric detection improved the platform's accuracy and reliability. The system also demonstrated excellent recovery performance in detecting AChE in complex serum samples. The proposed dual-mode sensing platform provides a sensitive, reliable, and robust tool for AChE detection, with promising applications in early diagnosis and treatment monitoring of neurodegenerative diseases.

The abnormal variation of phosphorylation can lead to many human diseases, such as Alzheimer's disease and cancer. Because of its high throughput and rapidity, mass spectrometry (MS)-based method has been widely used to characterize phosphopeptides/phosphoproteins in complex biological samples. However, the direct MS analysis for phosphopeptides is still a challenging task due to the complexity of biological samples and the signal suppression of abundant non-phosphopeptides. Therefore, an efficient enrichment platform for low-abundance phosphopeptide capture and detection is in great demand.

The tumor microenvironment (TME) refers to the complex ecological system surrounding tumor cells, which is intimately associated with regulating tumor cell growth, invasive behavior, and metastatic capacity. Hence, in situ imaging of related bioactivity with resolution in the TME is critical for early cancer detection and accurate diagnosis. In recent years, fluorescence imaging technology has become a widely used tool in TME research due to its non-invasive nature, high spatiotemporal resolution, and capability for real-time monitoring. Among these advancements, signal probes designed based on functional nucleic acids (FNAs) provide a promising and innovative toolkit for targeted imaging analysis of the TME.