Biological Detection Methods Research Articles

A high-quality fluorescence elimination dual-wavelength Raman method for biological detection and its application in cancer diagnosis

Raman-based methods offer a promising approach for in vivo biological detection. However, the fluorescence of biological samples will significantly affect Raman measurement accuracy. Moreover, due to the existence of excitation wavelength-dependent fluorescent molecules in biological tissues, especially porphyrin molecules, the fluorescence also exhibits significant wavelength dependence. To achieve high-quality Raman spectra of biological tissue, in this work we proposed a dual-wavelength Raman method. Two lasers with different wavelengths were used to excite optical signals in the same region, and the ordinary fluorescence and additional wavelength-dependent fluorescence in the biological samples could be eliminated by two-step normalization calibration; thus, the accuracy of Raman measurement was significantly enhanced. We applied this method to early cancer diagnosis and identified several molecules and structures worthy of attention in carcinogenesis for esophageal tissue, such as phenylalanine and the CC bonds of porphyrins. Normal, precancerous, and early cancer samples were successfully identified by the changes in biomolecules with associated degrees of malignancy. Thus, the imaging and diagnosis of indefinite tumors were realized, which verified the potential of the dual-wavelength Raman method in biological detection.

KGRACDA: A Model Based on Knowledge Graph from Recursion and Attention Aggregation for CircRNA-disease Association Prediction.

CircRNA is closely related to human disease, so it is important to predict circRNA-disease association (CDA). However, the traditional biological detection methods have high difficulty and low accuracy, and computational methods represented by deep learning ignore the ability of the model to explicitly extract local depth information of the CDA. We propose a model based on knowledge graph from recursion and attention aggregation for circRNA-disease association prediction (KGRACDA). This model combines explicit structural features and implicit embedding information of graphs, optimizing graph embedding vectors. First, we built large-scale, multi-source heterogeneous datasets and construct a knowledge graph of multiple RNAs and diseases. After that, we use a recursive method to build multi-hop subgraphs and optimize graph attention mechanism by gating mechanism, mining local depth information. At the same time, the model uses multi-head attention mechanism to balance global and local depth features of graphs, and generate CDA prediction scores. KGRACDA surpasses other methods by capturing local and global depth information related to CDA. We update an interactive web platform HNRBase v2.0, which visualizes circRNA data, and allows users to download data and predict CDA using model.

The clinical potential of 1,5-anhydroglucitol as biomarker in diabetes mellitus.

A crucial measure of diabetes management is to monitor blood glucose, which often requires continuous blood collection, leading to economic burden and discomfort. Blood glucose and glycated hemoglobin A1c serve as traditional indicators of glucose monitoring. But now glycated albumin, fructosamine, and 1,5-anhydroglucitol (1,5-AG) have been gaining more attention. 1,5-AG is a chemically stable monosaccharide that exists in the human body. Its serum concentration remains stable when blood glucose levels are normal. However, it decreases when blood glucose exceeds the renal glucose threshold. Studies have shown that 1.5-AG reflects blood glucose changes in 1 to 2 weeks; therefore, decreased levels of serum 1,5-AG can serve as a clinical indicator of short-term blood glucose disturbances. Recent studies have shown that 1,5-AG can be used not only for the screening and managing of diabetes but also for predicting diabetes-related adverse events and islet β cell function in prediabetic patients. In addition, saliva 1,5-AG demonstrates potential value in the screening and diagnosis of diabetes. This review focuses on the biological characteristics, detection methods, and clinical application of 1,5-AG to promote understanding and applicable research of 1,5-AG in the future.

Understanding the Antidepressant Mechanisms of Acupuncture: Targeting Hippocampal Neuroinflammation, Oxidative Stress, Neuroplasticity, and Apoptosis in CUMS Rats.

Depression is recognized globally as one of the most intractable diseases, and its complexity and diversity make treatment extremely challenging. Acupuncture has demonstrated beneficial effects in various psychiatric disorders. However, the underlying mechanisms of acupuncture's antidepressant action, particularly in depression, remain elusive. Therefore, this study aimed to investigate the effects of acupuncture on chronic unpredictability stress (CUMS)-induced depressive symptoms in rats and to further elucidate its underlying molecular mechanisms. All rats were exposed to CUMS of two stressors every day for 28 days, except for the control group. One hour before CUMS, rats were given a treatment with acupuncture, electroacupuncture, sham-acupuncture, or fluoxetine (2.1 mg/kg). Behavioral tests and biological detection methods were conducted in sequence to evaluate depression-like phenotype in rats. The findings of this study demonstrate that acupuncture therapy effectively ameliorated depression-like behavior induced by CUMS in rats. Additionally, acupuncture exerted a restorative effect on the alterations induced by CUMS in the levels of malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), brain-derived neurotrophic factor (BDNF), cyclic AMP response element-binding protein (CREB), postsynaptic density95 (PSD95), gamma-aminobutyric acid (GABA), and acetylcholine (ACh). Additionally, our findings indicate that acupuncture also modulates the ERK and Caspase-3 apoptotic pathways in the hippocampus of CUMS rats. This study suggests that acupuncture may play a potential preventive role by regulating hippocampal neuroinflammatory response, levels of oxidative stress, apoptotic processes, and enhancing synaptic plasticity.

Reverse iontophoresis sensing electrode for joint detection of pH, NH4+, and lactic acid

With the rapid development of integrated electronic circuit technology, biological fluid-electricity integrated detection systems have gradually become a research hotspot. Researchers have integrated biological fluid electrical detection methods into circuits and developed small, low-power, portable detection devices equipped with sensing electrodes, which can achieve continuous monitoring of human health parameters. In recent years, the integration of reverse iontophoresis technology that can be used to extract body fluids with electrochemical sensors has opened up the possibility of flexible, portable biochemical sensing with excellent detection sensitivity. Herein, we present a method for extracting biofluids based on reverse iontophoresis technology, coupled with electrochemical sensing techniques for the simultaneous detection of various biomarkers in body fluids. The multi-channel sensing electrode was modified layer by layer using nitrogen-doped graphene (N-Gr), ion-selective membrane, or lactate oxidase for rapid and sensitive detection of pH (3–8), ammonium ion (NH4+) (0.1 mM–50 mM), and lactic acid (1 mM–50 mM). Subsequently, an integrated system for electrical stimulation extraction and sensing analysis based on reverse iontophoresis was established and experimentally tested. Experimental results demonstrated that the multi-channel joint detection sensing electrode has excellent sensing linearity, specificity, repeatability, and long-term stability. Finally, a smartphone-based WeChat applet was developed, which can realize parameter setting, function selection, and result display of sensor detection. In this study, reverse iontophoresis technology was used to collect body fluids, and the important biomarkers pH, NH4+, and lactic acid in the body fluids were detected. Overall, this research presents an integrated detection system and a multi-channel sensing scheme for the detection of important biochemical markers in bodily fluids, thereby providing potential value for health monitoring applications.

Molybdenum disulfide quantum dots for rapid fluorescence detection of glutathione and ascorbic acid

A method for the specific detection of glutathione (GSH) and ascorbic acid (AA) using 3-aminophenylboronic acid functionalized molybdenum disulfide quantum dots (APBA-MoS2 QDs) was reported. The APBA-MoS2 QDs synthesized using the hydrothermal method showed the strongest fluorescence intensity at an emission wavelength of 375 nm with the optimum excitation wavelength of 320 nm. The detection method was constructed by the combination of the dynamic and static quenching and the inner filter effect (IFE) of APBA-MoS2 QDs by permanganate (MnO4−) and the rapid redox reaction (1 min) of MnO4− by GSH or AA. The concentrations of GSH and AA showed good linear ranges of 10–500 μM and 10–100 μM, respectively, with limits of detection (LOD) of 0.476 μM and 0.185 μM, respectively. The spiked recovery tests in human serum and fresh fruit samples showed that APBA-MoS2 QDs had significant potential in the construction of fluorescent biological detection methods.

Determination of monocrotaline and usaramine in rat plasma by UPLC-MS/MS and their pharmacokinetics

AbstractAn ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of monocrotaline and usaramine in rat plasma, to study the plasma drug concentration and pharmacokinetics, and to calculate the absolute bioavailability. The plasma was treated with acetonitrile and methanol (9:1, v/v) protein precipitation method. The chromatographic column was UPLC HSS T3 (50 mm × 2.1 mm, 1.7 μm), the mobile phase was methanol-water (containing 0.1% formic acid with 10 mM ammonium acetate in water), and the elution time was 4 min at a flow rate of 0.4 mL min−1. Electrospray ionization (ESI) positive ion mode was used for detection and multiple reaction monitoring (MRM) mode was used for quantitative analysis. Monocrotaline and usaramine were administered sublingual intravenously (iv) 1 mg kg−1 and orally (po) 5 mg kg−1, respectively, with 6 rats in each group, for a total of 24 rats. Then the pharmacokinetic differences in rats were evaluated. For the UPLC-MS/MS method, the calibration curve showed good linearity in the range of 2–2,000 ng mL−1, where r was greater than 0.99. The precision, accuracy, recovery, matrix effect and stability results were all consistent with the requirements of biological sample detection methods. to provide scientific experimental basis for the basic research The bioavailability of monocrotaline and usaramine in rat plasma was calculated, which was 43.5 and 19.5%, respectively.

Oriented surface imprinted 96-well microplate-based fluorescent biosensor for glycoprotein detection by boronate affinity sandwich assay

Glycoproteins perform vital functions in numerous biological processes and have important clinical implications. Many glycoproteins have been used as biomarkers and therapeutic targets for disease diagnosis. Due to low concentration of glycoprotein biomarkers and the presence of high-abundance interfering species in biological samples, a selective and sensitive detection method for glycoprotein is essential for real-world applications. In this study, we develop an oriented surface imprinted microplate-based fluorescent biosensor by boronate-affinity sandwich assay (BASA) for the specific, sensitive and high throughput determination of glycoproteins in complex samples. The structure of the BASA is based on sandwich formation between boronate affinity-oriented surface-imprinted microplates, target glycoproteins, and boronate affinity fluorescence probes. The imprinted microplates ensure the high specificity, high affinity and high throughput, while the fluorescence probes, consisting of boronic acid-modified CdTe QDs, provide high sensitivity. The proposed approach could exhibit a wide linear range of 1 ng/mL-105 ng/mL, with a low LOD of 0.528 ng/mL using horseradish peroxidase (HRP) as a model glycoprotein. As compared with traditional “turn off” fluorescent sensor, the developed “turn on” fluorescent sensor provided three orders of magnitude higher sensitivity at least. The fluorescent biosensor achieved average recoveries ranging from 96.8 % to 106.0 % in urine samples.

A Comprehensive Review on Circulating cfRNA in Plasma: Implications for Disease Diagnosis and Beyond.

Circulating cfRNA in plasma has emerged as a fascinating area of research with potential applications in disease diagnosis, monitoring, and personalized medicine. Circulating RNA sequencing technology allows for the non-invasive collection of important information about the expression of target genes, eliminating the need for biopsies. This comprehensive review aims to provide a detailed overview of the current knowledge and advancements in the study of plasma cfRNA, focusing on its diverse landscape and biological functions, detection methods, its diagnostic and prognostic potential in various diseases, challenges, and future perspectives.

The function of chemical folic acid in calibration methods and neurodevelopmental disorders.

Functional molecules have been attracting increasing attention in environmental and physiological studies. In particular, folic acid (FA) could be considered a key factor in estimating, adjusting, and making decisions in the treatment of neurodevelopmental disorders. It promotes the general significance and conceptual for considering FA molecular scientific research detections, which implies related advancement in both of biological structure and detection methods. Among these applications, the FA molecule acts as a coenzyme that incorporates carbon atoms and synthesizes purines and pyrimidines. Therefore, the calibration method has real applications and can be used as a sensing platform and for detection approaches, which conveys the internal relationship between the FA molecule and physiological characterization. This mini review briefly discusses multiple FA application fields and detection pathways and could supplement their utilization in anticipation of the onset of disease.

Surface Markers for the Identification of Cancer Stem Cells.

Cancer stem cells have genetic and functional characteristics which can turn them resistant to standard cancer therapeutic targets. Identification of these cells is challenging and is done mainly by detecting the expression of antigens specific to stem cells. Currently, there is a significant number of surface markers available which can detect cancer stem cells by directly targeting the specific antigens present in cells. These markers possess differential expression patterns and sub-localizations in cancer stem cells compared to nonneoplastic and somatic cells. In addition to these biomarkers, multiple analytical methods and techniques, including functional assays, cell sorting, filtration approaches, and xenotransplantation methods, are used to identify cancer stem cells. This chapter will overview the functional significance of cancer stem cells, their biological correlations, specific markers, and detection methods.

Application of Biological Methods and Remote Sensing in Marine Monitoring

Due to the serious side effects caused by poured nuclear sewage, marine monitoring ought to be paid much more attention than before. This article concludes two major general approaches, respectively biological methods and remote sensing. Biological methods contain specifically 4 different methods: ecological monitoring, biological monitoring, measurement of biological physiological and biochemical indicators, and determination of residual amounts of pollutants in the organism. Sensor monitoring technology based on marine organisms is a common technology, which desires marine life suitable for being sensor-equipped and well-developed tags to undergo successive monitoring. Biological detection methods are established by scientists in relevant fields to make up for the shortcomings of physical and chemical pollution detection. The remote sensing method, which includes a range of techniques such as satellite imagery, aerial photography, and sonar systems, offers a unique vantage point that allows us to observe and analyze large expanses of oceanic territory with unprecedented precision and efficiency. It can also be used to identify the sources of marine pollution, assess the impacts of marine pollution on marine ecosystems, monitor the effectiveness of pollution mitigation measures, and develop and implement policies to prevent marine pollution.

Development of a reporter gene-based assay for the bioactivity determination of rhLH pharmaceutical products

To establish an in vitro biological activity detection method for luteinizing hormone (LH), the hLHCGR-CREB-HEK293 cell line was constructed to stably express human luteinizing hormone/chorionic gonadotropin receptor (hLHCGR). After optimization, the rhLH starting working concentration was 800 mIU/mL with 4-fold serial dilutions, 10 concentrations and an incubation time of 5 h. The method was confirmed to be highly specific, with good accuracy, precision and linearity, meeting the needs of process research and release testing, and can be used as a routine detection method for LH biological activity. With the increasing demand for research and development of rhLH biologically similar drugs, establishing a stable and simple activity assay method to evaluate the biological activity of rhLH can provide technical support for quality control of rhLH products and powerful tools for comparability research of similar products.

An improved YOLOv5 method for clam seedlings biological feature detection under the microscope

In order to monitor the early growth status of clam seedlings and meet the demands of precision seedling cultivation in a factory setting, this study improves the YOLOv5s model and proposes a high-precision and lightweight method for detecting biological features of clam seedlings. Firstly, dataset of clam seedlings under the microscope was constructed. Secondly, we established a clam seedling detection model based on four different attention mechanisms and analyzed the differences in the features focused on by different attention mechanisms in clam seedling detection. The accuracy of the anchors was optimized using the K-means++ algorithm, and the Soft-NMS method was employed to address the issue of missed detections caused by dense stacking of clam seedlings. The proposed approach has the optimal comprehensive performance by comparing various lightweight networks. The results show that the AP-health, AP-death and mAP of this approach are 98.15%, 93.87% and 96.01%, which can meet the standard for high precision. With a transmission speed of 86 FPS, an average response time of 11.54 ms, and a model size of 27.59 MB, the approach satisfies the requirements for efficiency and portability. Finally, a biological feature detection software of clam seedlings was developed, which can automatically calculate phenotypic information such as the number of clam seedlings, survival rate, and average size. This paper provides a basis for real-time and accurate assessment of clam seedlings biological features while offering technical support for the automation and intelligence of clam seedlings production in factories.

Determination of Tenacissoside G, Tenacissoside H, and Tenacissoside I in Rat Plasma by UPLC-MS/MS and Their Pharmacokinetics.

An ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of tenacissoside G, tenacissoside H, and tenacissoside I in rat plasma. The rat plasma was treated with liquid-liquid extraction using ethyl acetate. The determination was performed on the UPLC HSS T3 column (50 mm × 2.1 mm, 1.8 μm) with a mobile phase consisting of acetonitrile-water (containing 0.1% formic acid) and gradient elution at a flow rate of 0.4 mL/min. Electrospray (ESI) positive ion mode detection and multireaction monitoring (MRM) quantitative analysis were performed. A total of 36 rats were given tenacissoside G, tenacissoside H, and tenacissoside I, respectively, orally (5 mg/kg) and intravenously (1 mg/kg), with 6 rats in each group, to evaluate the pharmacokinetic difference of tenacissoside G, tenacissoside H, and tenacissoside I in rats. The calibration curves showed good linearity in the range of 5-2000 ng/mL, where r was greater than 0.99. The results of precision, accuracy, recovery, matrix effect, and stability met the requirements of biological sample detection methods. The established UPLC-MS/MS method was successfully applied to pharmacokinetic studies of tenacissoside G, tenacissoside H, and tenacissoside I, and the bioavailability was 22.9%, 89.8%, and 9.4%, respectively.

Zero-Carbon and Carbon-Neutral Fuels: A Review of Combustion Products and Cytotoxicity

The use of zero-carbon and carbon-neutral fuels reduces emissions of conventional pollutants, but their emissions can be toxic and have various adverse effects on human health. This article reviews the possible combustion products of zero-carbon and carbon-neutral fuels, as well as their cytotoxic effects and potential health risks. At the same time, the review outlines biological models and toxicity detection methods commonly used in pollutant toxicity studies. Metals, nitrogen oxides (NOX), and ammonia (NH3) emitted from the combustion of metal fuels, hydrogen fuels, and ammonia fuels in zero-carbon fuels are harmful to human health. Exhaust emissions from carbon-neutral fuels, particularly biodiesel, and their blends with gasoline/diesel are cytotoxic, leading to severe cellular damage, such as oxidative damage, inflammatory responses, DNA damage, cell death, or apoptosis. Moreover, the normal function of the human body’s respiratory, cardiovascular, immune, digestive, urinary, and nervous systems may also be impacted by these fuel emissions according to cytotoxic research. Cytotoxicity of fuel combustion products is usually related to the fuel type, time, dose, and cell line used in the experiment. This review provides some ideas for the exhaust emission management of zero-carbon and carbon-neutral fuels and human health assessment. It also presents a theoretical and experimental basis for further research, including in vivo experiments.

The effect of acupuncture on lateral habenular nucleus and intestinal microflora in depression model rats

BackgroundDepression is a severe emotional condition that significantly affects the quality of life. Acupuncture exerts preventive effects on depression in rats with post-chronic unpredictable mild stress (CUMS). Methods The study involved chronic unpredictable mild stress (CUMS) depression model mice to administer acupuncture as a preventative measure to investigate the mechanism of acupuncture's antidepressant and observe the effect of acupuncture on impact via the Lateral Habenula (LHb) and Gut-Liver-Brain Axis. The researcher investigated molecules correlating with a nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway and assessed inflammation in the LHb and liver. In addition, 16 S rDNA bioinformatics study revealed the quantity and variety of gut microbiota. Rats were randomly divided into five groups: control (CON), CUMS, CUMS + acupuncture (AP), CUMS + fluoxetine (FX) and CUMS + N(G) -nitro -L- arginine methyl ester (LNAME) group. Except for the CON group, other rats were exposed to CUMS condition for 28 days. Simultaneously, manual acupuncture (at Fengfu and Shangxing acupoints, once every other day) and fluoxetine gavage (2.1 mg/kg, 0.21 mg/mL, daily) were conducted to the groups of AP and FX, respectively, after stressors. Rats in LNAME group were treated with LNAME normal saline (10 mg/kg, 1 mg/mL, i.p.) solution. Behavioural tests and biological detection methods were conducted sequentially to evaluate depressionlike phenotype in rats. ResultsThe results showed CUMS induced depression-like behaviours, hyper-activation of NO/cGMP signaling pathway, inflammation in serum, LHb and liver, and dysbiosis of the gut microbiota. These changes could be prevented and ameliorated by acupuncture to varying extents. ConclusionAcupuncture prevented and attenuated depression-like phenotype induced by CUMS, possibly via regulating the NO/cGMP signaling pathway and thus improving inflammation in serum, LHb and liver, and gut microbiota dysbiosis. In addition, these can be evidence of the existence of the gut-liver-brain axis.

Age-groups classification of Irrawaddy dolphins based on dorsal fin geometric morphological features

The Irrawaddy dolphin is a species at the top of the food chain, but its population is currently endangered. Hence, the critical need to take effective conservation measures for Irrawaddy dolphins, particularly in population reproduction. The age structure of Irrawaddy dolphins can be studied to understand the population reproduction status and provide reference for formulating scientific conservation measures. Unlike age estimation methods based on invasive biological tissue detection and individual biological target detection methods based on audio features or image pixel features, this paper proposes the method to extract the Irrawaddy dolphin's dorsal fin geometric morphological features for classifying age-groups by using non-uniform B-spline. Firstly, this method uses non-uniform B-spline method to fit the dorsal fin edge curve of Irrawaddy dolphins. Then the curve is resolved to extract its derivatives, curvature and other features that describe its geometry morphology. Finally, the random forest (RF) was supplemented to classify Irrawaddy dolphins into juvenile, middle-aged and older. Experiments show that the method proposed in this paper could effectively classify older dolphins to an accuracy of 87.36%. The method proposed in this paper is non-invasive not only can help analyze the Irrawaddy dolphin's development trajectory, understand the reproductive maturity, fecundity rates, and survival rates, but also can help understand the changes of the regional ecological environment and protect the aquatic diversity.

The trichotomy of HER2 expression confers new insights into the understanding and managing for breast cancer stratified by HER2 status.

Human epidermal growth factor receptor 2 (HER2) is a tyrosine kinase receptor that plays a carcinogenic role in breast cancer (BC) through gene amplification, mutation, or overexpression. Traditional methods of HER2 detection were divided into positive (immunohistochemistry (IHC) 3+/fluorescence in situ hybridization (FISH) amplification) and negative (IHC 2+/FISH-, IHC 1+, IHC 0) according to the dichotomy method. Anti-HER2-targeted therapies, such as trastuzumab and pertuzumab, have significantly improved the prognosis of HER2-positive patients. However, up to 75% to 85% of patients remain HER2-negative. In recent years, with the rapid development of molecular biology, gene detection technology, targeted therapy, and immunotherapy, researchers have actively explored the clinicopathological characteristics, molecular biological characteristics, treatment methods, and HER2 detection methods of HER2-low/zero breast cancer. With the clinical efficacy of new anti-HER2 targeted drugs, accurate classification of breast cancer is very important for the treatment choice. Therefore, the following review summarizes the necessity of developing HER2 detection methods, and the clinicopathological and drug treatment characteristics of patients with HER2-low/zero, to light the dawn of the treatment of breast cancer patients with HER2-low/zero expression.

Application of Luminescent Bacteria Bioassay in the Detection of Pollutants in Soil

The luminescent bacteria bioassay has been commonly used in the detection of environmental pollutants. Compared with traditional chemical and other biological detection methods, the luminescent bacteria bioassay has many demonstrated advantages such as a sensitive response, low cost, high efficiency, and environmental friendliness. The traditional luminescent bacteria bioassay has poor reproducibility and cannot achieve undisturbed soil testing, and the use of leach liquor also affects the results. This paper reviews the research progress and existing issues for the traditional luminescent bacteria bioassay used in the detection of soil pollutants. The luminescence mechanisms and detection principles of three commonly used luminescent bacteria, i.e., Vibrio fischeri, Photobacterium phosphoreum, and Vibrio qinghaiensis, are discussed and compared. In addition, two new luminescent bacteria bioassays are introduced to detect soil pollutants. One method is based on recombinant luminescent bacteria obtained with a gene-modification technique. This method can realize specific detection and enhance sensitivity, but it still cannot achieve undisturbed soil detection. The other method involves using magnetic nanoparticle (MNP)-based biosensors made from luminescent bacteria and MNPs. It can realize the accurate detection of the biological toxicity of the combined pollutants in soil without disturbing the soil’s integrity. This study shows that MNP-based biosensors have good application prospects in soil pollution detection, but the mechanism behind their utility still needs to be investigated to realize their popularization and application.