Internal standard optimization advances sensitivity and robustness of ratiometric detection method.

We design a p-aminothiophenol (pATP) modified Au/ITO chip to determine nitrite ions in lake water by a ratiometric surface-enhanced Raman scattering (SERS) method based on nitrite ions triggering the transformation of pATP to p,p'-dimercaptoazobenzene (DMAB). Intriguingly, by using the SERS peak (at 1008 cm-1) from benzoic ring deforming as an internal standard instead of the traditional peak at 1080 cm-1, the detection sensitivity of the method was improved 10 times.

Cat-scratch disease manifesting as uveitis and binocular fundus nodular lesions: a case report.

BACKGROUND: Cat-scratch disease typically presents with various ocular manifestations such as uveitis, vitritis, retinitis, retinochoroiditis, and optic neuritis. However, fundus nodular lesions was rarely reported. In our study, we reported a case of Cat-Scratch disease with binocular fundus nodular lesions. CASE PRESENTATION: An 11-year old male presented with uveitis in the right eye and bilateral fundus nodular lesions after indirect contact with unvaccinated cats. Comprehensive ancillary examinations including wide-angle fundus photography, ultrasonography, fluorescein fundus angiography, optical coherence tomography, and orbital magnetic resonance imaging were performed to elucidate the multidimensional features of the binocular lesions. Metagenomics next-generation sequencing was utilized to confirm the diagnosis of Cat-scratch disease. The patient's condition showed improvement after a 6-month combination treatment regimen involving systemic administration of doxycycline hyclate and methylprednisolone tablets, as well as local application of mydriatic and corticosteroid eye drops. CONCLUSIONS: We firstly reported a case of Cat-scratch disease presenting simultaneously with uveitis and fundus nodular lesions caused by Bartonella henselae infection in a child. Timely diagnosis and treatment with antibiotics and corticosteroids showed promising outcomes for the prognosis of these ocular disorders.

[Clinical, genetic, and pathological analysis in 165 children with disorders of sex development]. / 165ä¾‹å„¿ç«¥æ€§å‘è‚²å¼‚å¸¸ç–¾ç— çš„ä¸´åºŠã€é—ä¼ å­¦ä¸Žç— ç†å­¦åˆ†æž.

OBJECTIVES: To investigate the clinical phenotypes, genetic characteristics, and pathological features of children with disorders of sex development (DSD). METHODS: A retrospective analysis was conducted on epidemiological, clinical phenotype, chromosomal karyotype, gonadal pathology, and genotype data of 165 hospitalized children with DSD at Children's Hospital of Hebei Province and Tangshan Maternal and Child Health Hospital from August 2008 to December 2022. RESULTS: Among the 165 children with DSD, common presenting symptoms were short stature (62/165, 37.6%), clitoromegaly (33/165, 20.0%), cryptorchidism (28/165, 17.0%), hypospadias (24/165, 14.5%), and skin pigmentation abnormalities/exteriorized pigmented labia majora (19/165, 11.5%). Chromosomal karyotype analysis was performed on 127 cases, revealing 36 cases (28.3%) of 46,XX DSD, 34 cases (26.8%) of 46,XY DSD, and 57 cases (44.9%) of sex chromosome abnormalities. Among the sex chromosome abnormal karyotypes, the 45,X karyotype (11/57, 19%) and 45,X/other karyotype mosaicism (36/57, 63%) were more common. Sixteen children underwent histopathological biopsy of gonadal tissues, resulting in retrieval of 25 gonadal tissues. The gonadal tissue biopsies revealed 3 cases of testes, 3 cases of dysplastic testes, 6 cases of ovaries, 11 cases of ovotestes, and 1 case each of streak gonad and agenesis of gonads. Genetic testing identified pathogenic/likely pathogenic variants in 23 cases (23/36, 64%), including 12 cases of 21-hydroxylase deficiency congenital adrenal hyperplasia caused by CYP21A2 pathogenic variants. CONCLUSIONS: Short stature, clitoromegaly, cryptorchidism, hypospadias, and skin pigmentation abnormalities are common phenotypes in children with DSD. 45,X/other karyotype mosaicism and CYP21A2 compound heterozygous variants are major etiological factors in children with DSD. The most commonly observed gonadal histopathology in children with DSD includes ovotestes, ovaries, and testes/dysgenetic testes.

SERS determination of dopamine using metal-organic frameworks decorated with Ag/Au noble metal nanoparticle composite after azo derivatization with p-aminothiophenol.

A specific surface-enhanced Raman scattering (SERS) assay for dopamine (DA) based on an azo derivatization reaction is proposed for the first time by preparation of p-aminothiophenol (PATP)-modified composite SERS substrate, composed of metal-organic framework (MIL-101) decorated with Au and Ag nanoparticles. As the result, the SERS method for detection of the azo reaction between PATP and DA exhibits superior sensitivity, selectivity, and stability. A reasonable linearity in the range 10-6 to 10-10 mol∙L-1 is achieved, and the limit of detection is 1.2 × 10-12 mol∙L-1. The reactive SERS assay is free from interference in complex physiological fluid. The feasibility of the proposed SERS method for the detection of DA levels in fetal bovine serum (FBS) samples and human serum samples is validated by HPLC-MS methods, displaying promising application potential in early disease diagnosis.

Enzyme-Assist-Interference-Free Strategy for Raman Selective Determination of Sialic Acid.

Abnormal physiological levels of sialic acid (SA) could be used to diagnosis cancer progression stages. In this work, we describe an enzyme-assist-interference-free strategy for Raman selective determination of SA in serum. First, we assemble gold nanoparticles (Au NPs) onto the indium tin oxide glass (ITO) to construct an ITO/Au two-dimension substrate. Through modification of 4-mercaptoboric acid (4-MPBA) onto the surface of ITO/Au, the SA response plate is prepared due to the reversible esterification bond. In this strategy, a sandwich structure is rationally designed as ITO/Au/4-MPBA/SA/4-MPBA/Au to enhance the Raman scattering. The Raman detection linear concentration of SA ranged from 2.5 × 10-7 to 1.5 × 10-6 M, and a limit of detection about 1.2 × 10-7 M could be achieved. Considering the presence of glucose (Glu) in physiological fluid, we introduce glucose oxidase to remove the interference from Glu and realize the accurate determination of SA. The proposed novel Raman rapid method provides an ultrasensitive and interference-free protocol for the early diagnosis of cancer.

A flexible carbon nanotube-modified poly(styrene-butadiene)-based dopamine sensor.

In this work, a multi-walled carbon nanotube-modified flexible poly(styrene-butadiene) fiber membrane material was prepared for the sensitive and selective electrochemical detection of dopamine (DA) in human serum and DA injection. The flexible fiber membrane prepared by electrospinning technology is expected to realize its application in wearable devices. The obtained conductive film-based electrochemical sensor can effectively minimize interference caused by ascorbic acid and uric acid. Under the optimized experimental conditions of differential pulse voltammetry, DA gives a linear response in the range of 1-650 µM (R2 = 0.996). The detection limit of DA (signal-to noise ratio = 3) was determined to be 0.062 µM.

Fabrication of homogeneous waffle-like silver composite substrate for Raman determination of trace chloramphenicol.

Waffle-like anodized aluminum oxide homogeneously immobilized with Ag nanoparticles (AAO/Ag) is rationally designed and fabricated as surface-enhanced Raman scattering (SERS) substrate. The as-prepared SERS substrate is characterized with transmission electron microscope (TEM), scanning electron microscopy (SEM), UV-Vis spectrophotometer, and Fourier transform infrared spectrometer (FT-IR). The AAO/Ag substrate shows good uniformity of the Raman signals (RSD = 7.02%) due to waffle-like AAO supporting the well-dispersed Ag nanoparticles. For real application, the AAO/Ag substrate is used for rapid determination of chloramphenicol (CAP) in honey with low detection limit (4.0 × 10-9 mol L-1) and good linearity from 1.0 × 10-5 to 1.0 × 10-8 mol L-1 based on the SERS peak at 1348 cm-1. The better accumulation in the short pore path of AAO improves the target molecule approaching into the vicinity of hot spots of Ag nanoparticles. The high selectivity for CAP is attributed to the strong interaction between -NO2 group in CAP and the composite substrate. Schematic representation of the preparation of SERS substrate, AAO150/Ag10-5 composite nanoparticles, and antibiotic determination.

Preparation of Hydrophobic Film by Electrospinning for Rapid SERS Detection of Trace Triazophos.

For real application, it is an urgent demand to fabricate stable and flexible surface-enhanced Raman scattering (SERS) substrates with high enhancement factors in a large-scale and facile way. Herein, by using the electrospinning technique, a hydrophobic and flexible poly(styrene-co-butadiene) (SB) fibrous membrane is obtained, which is beneficial for modification of silver nanoparticles (Ag NPs) colloid in a small region and then formation of more "hot spots" by drying; the final SERS substrate is designated as Ag/SB. Hydrophobic Ag/SB can efficiently capture heterocyclic molecules into the vicinity of hot spots of Ag NPs. Such Ag/SB films are used to quantitatively detect trace triazophos residue on fruit peels or in the juice, and the limit of detection (LOD) of 2.5 × 10-8 M is achieved. Ag/SB films possess a capability to resist heat. As a case, 6-mercaptopurine (6MP) that just barely dissolves in 90 °C water is picked for conducting Ag/SB-film-based experiments.

Thiol-Disulfide Exchange Reaction for Cellular Glutathione Detection with Surface-Enhanced Raman Scattering.

Abnormal glutathione (GSH) levels are related to several diseases, including cancer. In this paper, surface-enhanced Raman spectroscopy (SERS) based on a 2-(pyridyldithio)ethylamine (PDEA)-modified AgNPs@Si wafer is proposed for intracellular GSH detection. PDEA plays multifunctional roles in the method, including the participation in the thiol-disulfide exchange reaction and the contribution of a great SERS signal reporter. With the addition of GSH, the disulfide bond of PDEA will be broken, releasing the pyridine ring in PDEA and resulting in a signal-off SERS response. The developed specific reaction-based SERS assay can detect GSH levels as low as 2.5 × 10-7 M. Subsequently, we employed this method to evaluate cellular GSH levels and found that the levels in cancer cells are higher than those in normal cells. The PDEA-modified AgNPs@Si-based SERS protocol demonstrates good selectivity and high sensitivity, as well as robustness, which is suitable to evaluate cellular GSH levels.

Surface enhanced Raman detection of the colon cancer biomarker cytidine by using magnetized nanoparticles of the type Fe3O4/Au/Ag.

Cytidine is regarded as an early marker of colon cancer. The authors describe a surface enhanced Raman scattering (SERS) technique to detect trace levels of cytidine in urine. The Raman band at 784 cm-1 can be acquired best. Compared to earlier methods, an improvement in detection sensitivity by a factor of 6.2 × 105 is achieved by using a magnetically induced method in which cytidine is captured in the vicinity of the SERS hot spots of the type Fe3O4/Au/Ag. Cytidine can be quantified at 1 nM levels by this method which is simple and reliable. Graphical Abstract Clusters consisting of magnetite (Fe3O4) nanoparticles, gold nanoparticles and silver nanoparticles were prepared and used in a SERS based method for detection of cytidine in urine by using magnetic improvement. The lowest detectable concentration of cytidine are at the nM level.

Photoelectrochemical determination of the activity of M.SssI methyltransferase, and a method for inhibitor screening.

A photoelectrochemical (PEC) method is described for the determination of the activity of M.SssI methyltransferase (MTase). The assay relies on enzyme-linkage reactions and a DNA intercalator Ru(bpy)2(dppz)2+ (where bpy is 2,2'-bipyridine, and dppz is dipyrido[3,2-a:2',3'-c]phenazine) which both serves as a PEC signal. The PEC electrode was obtained by immobilizing 5'-amino modified DNA strands (containing the methylation recognition site 5'-CCGG-3') on a polyethylenimine (PEI) coated ITO/SnO2 electrode with glutaraldehyde as crosslinking agent. In the presence of MTase and S-adenosyl-L-methionine, the 5'-CCGG-3' sequence in the DNA on the electrode is methylated. This protects the DNA strands from the shear of the methylation-sensitive restriction endonuclease HpaII. Consequently, more intact DNA strands remain on the surface of the electrode, providing more sites for Ru(bpy)2(dppz)2+ binding which in turn results in a high PEC response. The result demonstrates that the photocurrent increases linearly with the activity of MTase from 5 to 80 U·mL-1, and the limit of detection is 0.45 U·mL-1. The other MTases does not enhance the photocurrent, suggesting good selectivity of the assay. The method was also applied to rapid evaluate and screen the inhibitors of MTase. This strategy can be utilized to determinate the activity of other DNA MTases with specific DNA sequence. Graphical abstract Schematic presentation of a photoelectrochemical assay based on enzyme-linkage reactions and a photo electrochemical probe combined with the oxalic acid involved cyclic amplification system for the determination of methyltransferase activity.

The prevalence of hepatitis B infection in central China: An adult population-based serological survey of a large sample size.

The objective of this study was to determine the prevalence of serum HBsAg among adults in Henan province, China. A provincial serosurvey of people aged 18-74 years was conducted and individuals for analysis were selected using a multistage stratified random cluster sampling method. A 5-ml serum sample from each person was collected and the serum was tested for the presence of HBsAg, HBcAb, HBsAb, HBeAg, and HBeAb by ELISA. Multivariate analysis was used to identify factors associated with the prevalence of HBsAg. A total of 16,685 residents, aged 18-74 years who resided in 60 communities or villages in Henan, China were surveyed. Among the eligible study population, 642 HBsAg positive cases (3.7%) were found. The prevalence increased steadily from 3.1% among population aged 18-35 years old, to 5.1% among population aged 55-74 years. Accordingly, HBsAb declined steadily from 53.4% to 24.7%. Six hundred and forty-two positive HBsAg blood samples were screened for the HBeAg and HBeAb. HBeAg prevalence declined steadily from 27.7% among population age 18-35 years old to 6.8% among population age 55-74 years old. The single factor analysis was used for 16 possible risk factors and multivariate analysis showed that five risk factors were significantly related to HBV infection. HBV infection is a serious public health problem among adult population in Henan. Strengthening administrative regulations of medical practices, especially in rural areas, and providing health education propaganda to the public of HBV infection should be given more attention on public health policy. J. Med. Virol. 89:450-457, 2017. © 2016 Wiley Periodicals, Inc.

[Effects of Yiqi Bushen Experimental Recipe on mRNA Expressions of foxp3, STAT5, and NF-KB in Decidua Tissue of Natural Abortion Model Mice].

Objective To observe the effects of Yiqi Bushen Experimental Recipe (YBER, a rec- ipe for benefiting qi and Shen supplementing) on mRNA expression of foxp3 in splenic CD4 + CD25 Treg cells and mRNA expressions of foxp3, STAT5, and NF-κB in decidua tissue of natural abortion (NA) model mice. Methods Female CBA/J mice were caged and mated to male DBA/2 mice or male BALB/c mice in 2:1 for NA model. Pregnant CBA/J mice were randomly divided into 5 groups, i.e., the negative control group, the positive control group, high, middle, and low dose YBER groups, 10 in each group. Mice in the NS control group were administered with normal saline by gastrogavage from day 0 to their death, 10 mg/kg, once per day. Mice in the positive control group were administered with Cyclosporine A solution by gastrogavage on the 4th day of pregnancy. YBER (48, 24, 12 g/kg) was respectively administered to mice in the 3 dose YBER groups by gastrogavage from day 0 to their death, once per day. Preg- nant mice were sacrificed at day 9 and 14, and fresh spleens were taken out for extracting Treg cells. Dcidua tissues were collected and stored in -80 °C for frozen. Splenic CD4 + cells CD25 + were isolated and purified by magnetic bead. The purity of CD4 + cells CD25 + was identified by flow cytometry (FCM) before and after magnetic bead. mRNA expressions of foxp3, STAT5A, STAT5B, and NF-KB in decidua tissue were analyzed by RT-PCR. Results The purity of CD4 Treg CD25 could arrived at 88% plus. Its activity could be over 95% after trypan blue test. The average ratio of CD4 CD25+/CD4 was 13. 20% before purified isolation, while it was 91. 43% after purified isolation. Compared with the negative control group, foxp3 mRNA expression level in Treg cells was obviously elevated in the positive control group and the high dose YBER group (P <0. 05). foxp3 mRNA expression level in Treg cells was obviously ele- vated more in the high dose YBER group than in the middle dose YBER group and the low dose YBER group (P <0.05). Compared with the negative control group, mRNA expression levels of foxp3 and STAT5B in decidua tissue increased in the positive control group, high and middle dose YBER groups (P <0. 05). mRNA expression level of STAT5A increased in the positive control group and the high dose YBER group at day 9 and 14; as well as in the middle dose YBER group (P <0. 05, P <0. 01). NF-κB mR- NA expression level in decidua tissue was reduced in the positive control group and 3 dose YBER groups (P<0. 01). Conclusion YBER could up-regulate the expression of foxp3 mRNA in splenic CD4 + CD25 + Treg cells and mRNA expressions of foxp3 and STAT5 in decidua tissues of NA model mice, down-regulate NF-κB mRNA expression in maternal-fetal interface, and promote the maintenance of immune tolerance state.

Mitigation of retinol-induced skin irritation by physiologic lipids: Evidence from patch testing.

BACKGROUND: There is a dearth of effective treatments to counter retinol-induced skin irritation. OBJECTIVE: This study aimed to investigate the efficacy of three potential mitigants: (i) phytosteryl/octyldodecyl lauroyl glutamate (PLG), (ii) a physiologic lipid mixture (PLM) comprised of ceramide three and cholesterol, and (iii) niacinamide, in ameliorating irritation instigated by retinol. METHODS: An occlusive human patch test, spanning 5 days, was undertaken on 18 Chinese participants aged between 23 and 40. It was designed as a randomized, double-blind, and vehicle-controlled study. Clinician erythema assessment (CEA) and instrumental evaluations were employed pre and post-test. Subsequently, a 4-week consumer in-use test, randomized and double-blind in nature, was executed to substantiate the soothing effects of PLG. RESULTS: Data from CEA and bioengineering assessments revealed that, in comparison to the vehicle control, both 2% PLG and 5% PLM notably curbed retinol-induced skin erythema and inflammation. Notably, PLG outperformed PLM. Conversely, 3% niacinamide did not offer relief against retinol-induced discomfort. The subsequent consumer in-use test affirmed that treatments with 2% PLG were better tolerated than those with the vehicle alone. CONCLUSION: To the best of our knowledge, this study represents the first confirmation that physiologic lipids effectively mitigate retinol-induced irritation. Given their capacity to counter retinol-induced irritation, physiologic lipids, particularly PLG, are recommended for incorporation in retinol regimens. Additionally, the Visia-CR a* value can serve as a robust objective measure for interpreting patch test outcomes.

Rapid detection of I- in food samples by cholesteric chiral artificial receptor L5.

In this study, a cholesteric chiral artificial receptor L5 was synthesized and a rapid method for the detection of iodide ions was established. The addition of L5 to Ethanol/Tris (V/V = 1/1, pH 7.0) solution changed the iodide ion solution from colorless to light yellow to achieve the naked-eye identification. The identification pattern was confirmed with the help of spectral titration, job test and anti-interference test, and it was found that L5 bound to iodide ion in a 1:1 coordination ratio. It was also confirmed that the hydrogen atoms of the amide group of MeO-PhCONH in L5 and the hydrogen atoms of -COOCH3 in L5 interacted with I- through hydrogen bonding force by 1H NMR titration, infrared spectroscopy, scanning electron microscopy and computer molecular simulation. The detection limit of L5 was 0.025 µmol/L, which could efficiently and conveniently detect iodide ions in food samples.

O-phthalaldehyde assisted surface enhanced Raman spectroscopy selective determination of trace homocysteine in serum.

High plasma homocysteine (Hcy) levels may indicate cardiovascular disease. However, sensitive and selective determination of Hcy remains a major challenge. Herein, we present a sensing strategy for Hcy by surface enhanced Raman scattering (SERS) method along with a specific reaction of o-phthalaldehyde (OPA) and Hcy. The obtained adduct 2-(1-carboxyl-3-thiopropyl)-1-isoindolinone (Hcy-OPA) can be directly detected by SERS using gold nanoparticles (Au NPs) as the substrate. The developed SERS method displays superior sensitivity (low detection limit of 2.50 × 10-12 mol L-1) with a broad linear range (5.00 × 10-10 -5.00 × 10-6 mol L-1). As a proof of real application, it can be used to detect Hcy in bovine serum samples with a concentration as low as 5.00 × 10-9 mol L-1, which is free from the interference of the other amino acids and glutathione.

Reactive Hydrogel Patch for SERS Detection of Environmental Formaldehyde.

The tobacco epidemic is a public health threat that has taken a heavy toll of lives around the globe each year. As one of the poison species from smoking, formaldehyde (FA) affects both the smokers and nearby persons who are exposed to second-hand smoke. Therefore, on-site tracking of FA exposure could evaluate the public environmental safety and mitigate the potential hazards. Herein, we first prepare SiO2-shelled AuAg alloy nanoparticles (AuAg@SiO2) and then embed AuAg@SiO2 within agarose hydrogel to construct the three-dimensional (3D) surface-enhanced Raman scattering (SERS) substrate. A reagent of 3-methyl-2-benzothiazolinone hydrazine (MBTH) with reaction activity toward FA is loaded in the 3D substrate to obtain the selective SERS patch (designated as M-hydrogel patch). Based on a marker Raman peak at 1273 cm-1 from the reaction product of MBTH-FA, the M-hydrogel patch is used to realize SERS detection of FA in smoke. A good linear relationship from 5 × 10-4 to 5 mg/m3 and a limit of detection (LOD) of 2.92 × 10-5 mg/m3 could be reached. While for detection of FA in aqueous, a linear range of 1 × 10-7-1 × 10-3 mg/mL with an LOD of 1.46 × 10-8 mg/mL could be achieved. As the real application, the proposed M-hydrogel patch could be placed anywhere indoor to SERS evaluate the spatial distribution of FA in tobacco smoke, which is in connection with the second-hand smoke effect on children and adults. Such M-hydrogel patch-based SERS assay is exerted for on-field detection of FA in water and furniture panels by using a portable Raman system, showing satisfactory selectivity and reproducibility.

PU.1 positively regulates GATA-1 expression in mast cells.

Coexpression of PU.1 and GATA-1 is required for proper specification of the mast cell lineage; however, in the myeloid and erythroid lineages, PU.1 and GATA-1 are functionally antagonistic. In this study, we report a transcriptional network in which PU.1 positively regulates GATA-1 expression in mast cell development. We isolated a variant mRNA isoform of GATA-1 in murine mast cells that is significantly upregulated during mast cell differentiation. This isoform contains an alternatively spliced first exon (IB) that is distinct from the first exon (IE) incorporated in the major erythroid mRNA transcript. In contrast to erythroid and megakaryocyte cells, in mast cells we show that PU.1 and GATA-2 predominantly occupy potential cis-regulatory elements in the IB exon region in vivo. Using reporter assays, we identify an enhancer flanking the IB exon that is activated by PU.1. Furthermore, we observe that in PU.1(-/-) fetal liver cells, low levels of the IE GATA-1 isoform is expressed, but the variant IB isoform is absent. Reintroduction of PU.1 restores variant IB isoform and upregulates total GATA-1 protein expression, which is concurrent with mast cell differentiation. Our results are consistent with a transcriptional hierarchy in which PU.1, possibly in concert with GATA-2, activates GATA-1 expression in mast cells in a pathway distinct from that seen in the erythroid and megakaryocytic lineages.

Beneficial effect of Bupleurum polysaccharides on autoimmune-prone MRL-lpr mice.

Systemic lupus erythematosus (SLE) is a chronic systemic autoimmune disease leading to inflammatory tissue damage in multiple organs. The crude polysaccharides (BPs) isolated from the roots of Bupleurum smithii var. parvifolium have anticomplementary activity and immunomodulatory functions on macrophages. To study its potential benefit on SLE, we examined effects of BPs on MRL-lpr mice, which have similar disease features to human SLE. MRL-lpr mice were treated orally with BPs 15, 30, or 60 mg kg⁻¹ day⁻¹ for 12 weeks and their SLE characteristics were evaluated. The results revealed that BPs elongated life span, improved kidney function, delayed lymphadenopathy, and reduced autoantibodies. It seemed to be mediated by inhibition of complement and macrophages activation and suppression of interferon-γ (IFN-γ) and interleukin-6 (IL-6) gene expression in the kidney. These results implicate that BPs may be an immunomodulator for the treatment of autoimmune diseases like SLE.

Detection strategies for superoxide anion: A review.

Reactive oxygen species (ROS) play an essential role in regulating various physiological functions of living organisms. Superoxide anion (O2-.), one kind of ROS, is the single-electron reduction product of oxygen molecules, which mainly exists in plants and animals, and is closely related to many inflammatory diseases. In the field of biomedicine, with the deepening understanding of superoxide anion, more and more detection methods have been developed. This review mainly introduces the detection techniques for superoxide anion in recent years.