Empirical Confirmation of Theoretical Predictions for Amorphous H/VO4 Cathodes: Advancing Durability and Efficiency in Zinc-Ion Batteries.

Advancing cathode materials is crucial for the broader application of aqueous zinc-ion batteries (ZIBs) in energy storage systems. This study presents amorphous H/VO4 (HVO), a novel cathode material engineered by substituting H+ for Mg2+ in Mg2VO4 (MgVO), designed to enhance performance of ZIBs. Initial exploration of MgVO through ab initio molecular dynamics (AIMD) simulations and density functional theory (DFT) calculations revealed a favorable Mg2+ and Zn2+ exchange mechanism. This mechanism notably reduces electrostatic interactions and facilitates ion diffusion within the host lattice. Building upon these findings, in this work, theoretical calculations analysis indicated that amorphous HVO offers a higher diffusion coefficient for Zn2+ ions and fewer electrostatic interactions compared to its crystalline MgVO precursor. Subsequent empirical validation is achieved by synthesizing amorphous HVO using a rapid ion-exchange process, effectively replacing Mg2+ with H+ ions. The synthesized amorphous HVO demonstrated 100% capacity retention after 18000 cycles at a current density of 2 A g-1 and exhibited exceptional rate performance. These findings underscore the significant potential of HVO cathodes to enhance the durability and efficiency of aqueous ZIBs, positioning them as promising candidates for future energy storage technologies.

Amorphous Structure Benefits in MgV2O4/V2O3 Composites for Zinc-Ion Storage: An Integration of Computational and Experimental Studies.

This study investigates the electrochemical properties of MgV2O4/V2O3 composites for Aqueous Zinc-Ion Batteries (AZIBs) using both Density Functional Theory (DFT) calculations and experimental validation. DFT analysis reveals significant electron mobility and reactivity at the MgV2O4/V2O3 interface, enhancing Zn2+ storage capabilities. This theoretical prediction is confirmed experimentally by synthesizing a novel MgV2O4/V2O3 composite that demonstrates superior electrochemical performance compared to pristine phases. Notably, the transition of the MgV2O4/V2O3 composite into an amorphous structure during electrochemical cycling is pivotal, providing enhanced diffusion pathways and increased conductivity. The composite delivers a consistent specific capacity of 330.2 mAh g-1 over 50 cycles at 0.1 A g-1 and maintains 152.7 mAh g-1 at an elevated current density of 20 A g-1 after 2000 cycles, validating the synergy between DFT insights and experimental outcomes, and underscoring the potential of amorphous structures in enhancing battery performance.

PM2.5 exposure and its interaction of oxidative balance score on ovarian cancer survival: A prospective cohort study.

Recent evidence advises particles with a diameter of 2.5 µm or less (PM2.5) might be a prognostic factor for ovarian cancer (OC) survival. The oxidative balance score (OBS) incorporates diet-lifestyle factors to estimate individuals' anti-oxidant exposure status which may be relevant to cancer prognosis. We aimed to investigate the roles of PM2.5, and OBS and their interaction in OC prognosis. 663 patients with OC were enrolled in the current study. Satellite-derived annual average exposures to PM2.5 based on patients' residential locations. The OBS was calculated based on 16 different diet-lifestyle components derived using an acknowledged self-reported questionnaire. The Cox regression model was performed to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs) for overall survival (OS). We also assessed the effect of modification between PM2.5 and OS by OBS via interaction terms. During a median follow-up of 37.57 (interquartile:35.27-40.17) months, 123 patients died. Compared to low-concentration PM2.5 exposure, high PM2.5 during 1 year before diagnosis was associated with worse OC survival (HR= 1.19, 95% CI = 1.01-1.42). We observed an improved OS with the highest compared with the lowest OBS (HR = 0.46, 95% CI = 0.27-0.79, P for trend < 0.05). Notably, we also found an additive interaction between low OBS and high exposure to PM2.5, with the corresponding associations of PM2.5 being more pronounced among participants with lower OBS (HR = 1.42, 95% CI = 1.09-1.86). PM2.5 may blunt OC survival, but high OBS represented an antioxidative performance that could alleviate the adverse association of PM2.5 and OS.

Determination of antihypertensive drugs irbesartan and doxazosin mesylate in healthcare products and urine samples using surface-enhanced Raman scattering.

In this paper, a surface-enhanced Raman scattering (SERS) strategy was constructed for the determination of antihypertensive drugs irbesartan (IRB) and doxazosin mesylate (DOX). ß-Cyclodextrin-capped silver nanoparticles (CD-AgNPs) are employed as SERS-active substrate. The introduction of ß-CD with hydrophobic cavity can capture drug molecules to form host-guest complex, making the drug molecules closer to the electromagnetic enhancement field of the AgNPs, thereby enhancing the SERS signal of drug molecules with low Raman cross-section. The vibrational modes of IRB and DOX are assigned by density functional theory calculations. The linear response from 3.0 × 10-7 to 1.0 × 10-5 mol L-1 for IRB and 3.0 × 10-7 to 2.0 × 10-5 mol L-1 for DOX and low limits of detection (LOD) 7.5 × 10-8 mol L-1 for IRB and 8.6 × 10-8 mol L-1 for DOX can be achieved. Meanwhile, this SERS approach can be applicable to determine IRB and DOX in commercial drug tablets, healthcare products, and human urine samples with recoveries of 90.8-115.7% and 90.0-113.5%, respectively, with relative standard deviation (RSD) less than 4.5%. This designed SERS strategy enables for the rapid determination of IRB and DOX in drug quality monitoring and illegal additives in healthcare products as well as clinical applications.

Wearable Inertial Measurement Unit Sensing System for Musculoskeletal Disorders Prevention in Construction.

Construction workers executing manual-intensive tasks are susceptible to musculoskeletal disorders (MSDs) due to overexposure to awkward postures. Automated posture recognition and assessment based on wearable sensor output can help reduce MSDs risks through early risk-factor detection. However, extant studies mainly focus on optimizing recognition models. There is a lack of studies exploring the design of a wearable sensing system that assesses the MSDs risks based on detected postures and then provides feedback for injury prevention. This study aims at investigating the design of an effective wearable MSDs prevention system. This study first proposes the design of a wearable inertial measurement unit (IMU) sensing system, then develops the prototype for end-user evaluation. Construction workers and managers evaluated a proposed system by interacting with wearable sensors and user interfaces (UIs), followed by an evaluation survey. The results suggest that wearable sensing is a promising approach for collecting motion data with low discomfort; posture-based MSDs risk assessment has a high potential in improving workers' safety awareness; and mobile- and cloud-based UIs can deliver the risk assessment information to end-users with ease. This research contributes to the design, development, and validation of wearable sensing-based injury prevention systems, which may be adapted to other labor-intensive occupations.

Heterologous expression of a GH3 ß-glucosidase from Neurospora crassa in Pichia pastoris with high purity and its application in the hydrolysis of soybean isoflavone glycosides.

Previous studies have shown isoflavone aglycones to have more biological effects than their counterparts, isoflavone glycones. Some ß-glucosidases can hydrolyze isoflavone glucosides to release aglycones, and discovery of these has attracted great interest. A glycoside hydrolase (GH) family 3 ß-glucosidase (bgl2) gene from Neurospora crassa was heterologously expressed in Pichia pastoris with high purity. The recombinant BGL2 enzyme displayed its highest activity at pH 5.0 and 60 °C, and had its maximum activity against p-nitrophenyl-ß-d-glucopyranoside (pNPG) (143.27 ± 4.79 U/mg), followed by cellobiose (74.99 ± 0.78 U/mg), gentiobiose (47.55 ± 0.15 U/mg), p-nitrophenyl-ß-d-cellobioside (pNPC) (40.07 ± 0.87 U/mg), cellotriose (12.31 ± 0.36 U/mg) and cellotetraose (9.04 ± 0.14 U/mg). The kinetic parameters of Km and Vmax were 0.21 ± 0.01 mM and 147.93 ± 2.77 µM/mg/min for pNPG. The purified enzyme showed a heightened ability to convert the major soybean isoflavone glycosides (daidzin, genistin and glycitin) into their corresponding aglycone forms (daidzien, genistein and glycitein). With this activity against soybean isoflavone glycosides, BGL2 shows great potential for applications in the food, animal feed, and pharmaceutical industries.

A C-terminal proline-rich sequence simultaneously broadens the optimal temperature and pH ranges and improves the catalytic efficiency of glycosyl hydrolase family 10 ruminal xylanases.

Efficient degradation of plant polysaccharides in rumen requires xylanolytic enzymes with a high catalytic capacity. In this study, a full-length xylanase gene (xynA) was retrieved from the sheep rumen. The deduced XynA sequence contains a putative signal peptide, a catalytic motif of glycoside hydrolase family 10 (GH10), and an extra C-terminal proline-rich sequence without a homolog. To determine its function, both mature XynA and its C terminus-truncated mutant, XynA-Tr, were expressed in Escherichia coli. The C-terminal oligopeptide had significant effects on the function and structure of XynA. Compared with XynA-Tr, XynA exhibited improved specific activity (12-fold) and catalytic efficiency (14-fold), a higher temperature optimum (50°C versus 45°C), and broader ranges of temperature and pH optima (pH 5.0 to 7.5 and 40 to 60°C versus pH 5.5 to 6.5 and 40 to 50°C). Moreover, XynA released more xylose than XynA-Tr when using beech wood xylan and wheat arabinoxylan as the substrate. The underlying mechanisms responsible for these changes were analyzed by substrate binding assay, circular dichroism (CD) spectroscopy, isothermal titration calorimetry (ITC), and xylooligosaccharide hydrolysis. XynA had no ability to bind to any of the tested soluble and insoluble polysaccharides. However, it contained more α helices and had a greater affinity and catalytic efficiency toward xylooligosaccharides, which benefited complete substrate degradation. Similar results were obtained when the C-terminal sequence was fused to another GH10 xylanase from sheep rumen. This study reveals an engineering strategy to improve the catalytic performance of enzymes.

Quinoline yellow acts as a novel amyloid fibrillation probe by using surface-enhanced Raman spectroscopy.

Protein amyloid fibrillation is linked to a wide range of neurodegenerative diseases. Protein oligomer is an intermediate substance in the process of fibrillation, which is neurotoxic and formed by the aggregation of protein molecules under physiological stress. Early detection of protein oligomers could make timely intervention of protein fibrillation related diseases. Therefore, it is crucial to develop efficient inhibitors and probes for monitoring amyloid fibril formation. In this study, we developed a novel amyloid inhibitor quinoline yellow (QY), which was proved to be effective in inhibiting insulin protein fibrillation as demonstrated by fluorescence, morphology characterization and circular dichroism. When QY binds to insulin, it exerts inhibitory effects on the nucleation process and effectively impedes the formation of fibrillar fibrils. In addition, we present the application of surface-enhanced Raman spectroscopy (SERS) as an extremely sensitive technique for identifying amyloid oligomers. The investigation employed the probe QY, which demonstrated a linear reaction for identifying oligomers in the concentration range of 1.0-58.0 µM. Impressively, it showcased an exceptionally sensitive detection threshold of 0.2 µM. And also illustrating the binding sites and interaction mechanisms between small molecules of QY and insulin by SERS. The aforementioned methodology was also employed for the identification of insulin oligomers in human serum samples. Thereby, the proposed approach presenting a promising avenue with extensive implications in the realms of pharmaceutical exploration and disease diagnosis.

Rewiring metabolic flux to simultaneously improve malate production and eliminate by-product succinate accumulation by Myceliophthora thermophila.

Although a high titre of malic acid is achieved by filamentous fungi, by-product succinic acid accumulation leads to a low yield of malic acid and is unfavourable for downstream processing. Herein, we conducted a series of metabolic rewiring strategies in a previously constructed Myceliophthora thermophila to successfully improve malate production and abolish succinic acid accumulation. First, a pyruvate carboxylase CgPYC variant with increased activity was obtained using a high-throughput system and introduced to improve malic acid synthesis. Subsequently, shifting metabolic flux to malate synthesis from mitochondrial metabolism by deleing mitochondrial carriers of pyruvate and malate, led to a 53.7% reduction in succinic acid accumulation. The acceleration of importing cytosolic succinic acid into the mitochondria for consumption further decreased succinic acid formation by 53.3%, to 2.12 g/L. Finally, the importer of succinic acid was discovered and used to eliminate by-product accumulation. In total, malic acid production was increased by 26.5%, relative to the start strain JG424, to 85.23 g/L and 89.02 g/L on glucose and Avicel, respectively, in the flasks. In a 5-L fermenter, the titre of malic acid reached 182.7 g/L using glucose and 115.8 g/L using raw corncob, without any by-product accumulation. This study would accelerate the industrial production of biobased malic acid from renewable plant biomass.

Carbon nanotubes integrated photonic barcodes in Herringbone Microfluidics for Multiplex Biomarker Quantification.

Early monitoring of cardiovascular disease (CVD) is crucial for its treatment and prognosis. Hence, highly specific and sensitive detection method is urgently needed. In this study, we propose a novel herringbone microfluid chip with aptamer functionalized core-shell photonic crystal (PhC) barcode integration for high throughput multiplex CVD detection. Based on the PhC derived from co-assembled carboxylated single-wall carbon nanotubes and silicon dioxide nanoparticles, we obtain core-shell PhC barcodes by hydrogel replicating and partially etching. These core-shell PhC barcodes not only retain the original structural colors coding element, but also fully expose a large number of carboxyl elements in the ore for the probe immobilization. We further combine the functionalized barcodes with herringbone groove microfluidic chip to elucidate its acceptability in testing clinical sample. It is demonstrated that the special design of microfluidic chip can significantly enhance fluid vortex resistance and contact frequency, improving the sample capture efficiency and detection sensitivity. These features indicate that our core-shell PhC barcodes-integrated herringbone microfluidic system possesses great potential for multiplex biomarker detection in clinical application.

Metagenomics-based gene exploration and biochemical characterization of novel glucoamylases and α-amylases in Daqu and Pu-erh tea microorganisms.

α-Amylases and glucoamylases play a crucial role in starch degradation for various industrial applications. Further exploration of novel α-amylases and glucoamylases with diverse enzymatic characteristics is necessary. In this study, metagenomics analysis revealed a high abundance of these enzymes in the microorganisms of Daqu and Pu-erh tea, identifying 271 glucoamylases and 232 α-amylases with significant sequence identity to known enzymes. Functional studies indicated that these enzymes have broad optimal temperatures (30 °C to 70 °C) and acidic or neutral pH optima. Additionally, two novel low-temperature glucoamylases and one novel low-temperature α-amylases were characterized, demonstrating potential for use in industries operating under low temperature conditions. Further analysis suggested that fewer molecular interactions and more flexible coli regions may contribute to their high activity at low temperatures. In summary, this study not only highlights the feasibility of exploring enzymes through metagenomic approaches, but also presents a library of novel and diverse α-amylases and glucoamylases for potential industrial applications.

Unsupervised domain adaptation multi-level adversarial learning-based crossing-domain retinal vessel segmentation.

BACKGROUND: The retinal vasculature, a crucial component of the human body, mirrors various illnesses such as cardiovascular disease, glaucoma, and retinopathy. Accurate segmentation of retinal vessels in funduscopic images is essential for diagnosing and understanding these conditions. However, existing segmentation models often struggle with images from different sources, making accurate segmentation in crossing-source fundus images challenging. METHODS: To address the crossing-source segmentation issues, this paper proposes a novel Multi-level Adversarial Learning and Pseudo-label Denoising-based Self-training Framework (MLAL&PDSF). Expanding on our previously proposed Multiscale Context Gating with Breakpoint and Spatial Dual Attention Network (MCG&BSA-Net), MLAL&PDSF introduces a multi-level adversarial network that operates at both the feature and image layers to align distributions between the target and source domains. Additionally, it employs a distance comparison technique to refine pseudo-labels generated during the self-training process. By comparing the distance between the pseudo-labels and the network predictions, the framework identifies and corrects inaccuracies, thus enhancing the accuracy of the fine vessel segmentation. RESULTS: We have conducted extensive validation and comparative experiments on the CHASEDB1, STARE, and HRF datasets to evaluate the efficacy of the MLAL&PDSF. The evaluation metrics included the area under the operating characteristic curve (AUC), sensitivity (SE), specificity (SP), accuracy (ACC), and balanced F-score (F1). The performance results from unsupervised domain adaptive segmentation are remarkable: for DRIVE to CHASEDB1, results are AUC: 0.9806, SE: 0.7400, SP: 0.9737, ACC: 0.9874, and F1: 0.8851; for DRIVE to STARE, results are AUC: 0.9827, SE: 0.7944, SP: 0.9651, ACC: 0.9826, and F1: 0.8326. CONCLUSION: These results demonstrate the effectiveness and robustness of MLAL&PDSF in achieving accurate segmentation results from crossing-domain retinal vessel datasets. The framework lays a solid foundation for further advancements in cross-domain segmentation and enhances the diagnosis and understanding of related diseases.

Metallofullerenol alleviates alcoholic liver damage via ROS clearance under static magnetic and electric fields.

Alcoholic liver disease (ALD) is a common chronic redox disease caused by increased alcohol consumption. Abstinence is a major challenge for people with alcohol dependence, and approved drugs have limited efficacy. Therefore, this study aimed to explore a new treatment strategy for ALD using ferroferric oxide endohedral fullerenol (Fe3O4@C60(OH)n) in combination with static magnetic and electric fields (sBE). The primary hepatocytes of 8-9-week-old female BALB/c mice were used to evaluate the efficacy of the proposed combination treatment. A mouse chronic binge ethanol feeding model was established to determine the alleviatory effect of Fe3O4@C60(OH)n on liver injury under sBE exposure. Furthermore, the ability of Fe3O4@C60(OH)n to eliminate •OH was evaluated. Alcohol-induced hepatocyte and mitochondrial damage were reversed in vitro. Additionally, the combination therapy reduced liver damage, alleviated oxidative stress by improving antioxidant levels, and effectively inhibited liver lipid accumulation in animal experiments. Here, we used a combination of magnetic derivatives of fullerenol and sBE to further improve the ROS clearance rate, thereby alleviating ALD. The developed combination treatment may effectively improve alcohol-induced liver damage and maintain redox balance without apparent toxicity, thereby enhancing therapy aimed at ALD and other redox diseases.

Comparative quantitative analysis of gene expression profiles of glycoside hydrolase family 10 xylanases in the sheep rumen during a feeding cycle.

Xylanase is a crucial hydrolytic enzyme that degrades plant polysaccharides in the rumen. To date, there is no information on the genetic composition and expression characteristics of ruminal xylanase during feeding cycles of ruminants. Here, the major xylanase of the glycoside hydrolase family 10 (GH 10) from the rumen of small-tail Han sheep was investigated during a feeding cycle. We identified 44 distinct GH 10 xylanase gene fragments at both the genomic and transcriptional levels. Comparison of their relative abundance showed that results from the evaluation of functional genes at the transcriptional level are more reliable indicators for understanding fluctuations in xylanase levels. The expression patterns of six xylanase genes, detected at all time points of the feeding cycle, were investigated; we observed a complex trend of gene expression over 24 h, revealing the dynamic expression of xylanases in the rumen. Further correlation analysis indicated that the rumen is a dynamic ecosystem where the transcript profiles of xylanase genes are closely related to ruminal conditions, especially rumen pH and bacterial population. Given the huge diversity and changing composition of enzymes over the entire rumen, this research provides valuable information for understanding the role of functional genes in the digestion of plant material.

Developing Liver Microphysiological Systems for Biomedical Applications.

Microphysiological systems (MPSs), also known as organ chips, are micro-units that integrate cells with diverse physical and biochemical environmental cues. In the field of liver MPSs, cellular components have advanced from simple planar cell cultures to more sophisticated 3D formations such as spheroids and organoids. Additionally, progress in microfluidic devices, bioprinting, engineering of matrix materials, and interdisciplinary technologies have significant promise for producing MPSs with biomimetic structures and functions. This review provides a comprehensive summary of biomimetic liver MPSs including their clinical applications and future developmental potential. First, the key components of liver MPSs, including the principal cell types and engineered structures utilized for cell cultivation, are briefly introduced. Subsequently, the biomedical applications of liver MPSs, including the creation of disease models, drug absorption, distribution, metabolism, excretion, and toxicity, are discussed. Finally, the challenges encountered by MPSs are summarized, and future research directions for their development are proposed.

Bi2O3/Gd2O3 Meta-Aerogel with Leaf-Inspired Nanotrap Array Enables Efficient X-Ray Absorption.

The increasing utilization of X-rays has generated a growing need for efficient shielding materials. However, the existing Pb-based materials suffer from a narrow X-ray absorbing range, high weight, and rigidity. Inspired by the natural leaf, which can efficiently absorb light through chlorophyll and carotenoids in confined cells, we engineer ultralight and superelastic nanofibrous Bi2O3/Gd2O3 meta-aerogels (BGAs) with X-ray nanotrap arrays by manipulating the 3D confined assembly of 1D Bi2O3 and Gd2O3 nanofibers. The BGAs can synergistically absorb X-ray photons from complementary energy ranges into the nanotraps and induce cyclic collisions with Bi2O3 and Gd2O3 nanofibers, maximizing the effective X-ray attenuation. The meta-aerogel exhibits the integrated performance of efficient X-ray shielding efficiency (60-83%, 16-90 keV), ultralow density (10 mg cm-3), and superelasticity. The production of these meta-aerogels presents an avenue for the development of next-generation X-ray protective materials and the resolution of X-ray imaging systems.

Role of adenosine A2a receptor in cancers and autoimmune diseases.

Adenosine receptors are P1 class of purinergic receptors that belong to G protein-coupled receptors. There are 4 subtypes of adenosine receptors, namely A1, A2A, A2B, and A3. A2AR has a high affinity for the ligand adenosine. Under pathological conditions or external stimuli, ATP is sequentially hydrolyzed to adenosine by CD39 and CD73. The combination of adenosine and A2AR can increase the concentration of cAMP and activate a series of downstream signaling pathways, and further playing the role of immunosuppression and promotion of tumor invasion. A2AR is expressed to some extent on various immune cells, where it is abnormally expressed on immune cells in cancers and autoimmune diseases. A2AR expression also correlates with disease progression. Inhibitors and agonists of A2AR may be potential new strategies for treatment of cancers and autoimmune diseases. We herein briefly reviewed the expression and distribution of A2AR, adenosine/A2AR signaling pathway, expression, and potential as a therapeutic target.

Cortical Organoid-on-a-Chip with Physiological Hypoxia for Investigating Tanshinone IIA-Induced Neural Differentiation.

Cortical organoids represent cutting-edge models for mimic human brain development during the early and even middle stage of pregnancy, while they often fail to recreate the complex microenvironmental factors, such as physiological hypoxia. Herein, to recapitulate fetal brain development, we propose a novel cortical organoid-on-a-chip with physiological hypoxia and further explore the effects of tanshinone IIA (Tan IIA) in neural differentiation. The microfluidic chip was designed with a micropillar array for the controlled and efficient generation of cortical organoids. With low oxygen, the generated cortical organoids could recapitulate key aspects of early-gestational human brain development. Compared to organoids in normoxic culturing condition, the promoted neurogenesis, synaptogenesis and neuronal maturation were observed in the present microsystem, suggesting the significance of physiological hypoxia in cortical development. Based on this model, we have found that Chinese herbal drug Tan IIA could promote neural differentiation and maturation, indicating its potential therapeutic effects on neurodevelopmental disorders as well as congenital neuropsychiatric diseases. These results indicate that the proposed biomimetic cortical organoid-on-a-chip model with physiological hypoxia can offer a promising platform to simulate prenatal environment, explore brain development, and screen natural neuroactive components.

Detoxification of Fumonisins by Three Novel Transaminases with Diverse Enzymatic Characteristics Coupled with Carboxylesterase.

Fumonisin (FB) is one of the most common mycotoxins contaminating feed and food, causing severe public health threat to human and animals worldwide. Until now, only several transaminases were found to reduce FB toxicity, thus, more fumonisin detoxification transaminases with excellent catalytic properties required urgent exploration for complex application conditions. Herein, through gene mining and enzymatic characterization, three novel fumonisin detoxification transaminases-FumTSTA, FumUPTA, FumPHTA-were identified, sharing only 61-74% sequence identity with reported fumonisin detoxification transaminases. Moreover, the recombinant proteins shared diverse pH reaction ranges, good pH stability and thermostability, and the recombinant protein yields were also improved by condition optimum. Furthermore, the final products were analyzed by liquid chromatography-mass spectrometry. This study provides ideal candidates for fumonisin detoxification and meets diverse required demands in food and feed industries.

SERS-based biosensor for detection of f-PSA%: Implications for the diagnosis of prostate cancer.

In diagnosing prostate cancer and distinguishing it from other prostate diseases, the ratio of the concentration of free prostate-specific antigen (f-PSA) to total prostate-specific antigen (t-PSA), i.e., (f-PSA%) is more accurate than the concentration of t-PSA alone. Immunoassay based on surface-enhanced Raman scattering (SERS) frequency shift has been proven to be particularly suitable for detecting large biomolecules with high reproducibility. Along similar lines, the present study developed a SERS-based biosensor that simultaneously detects t-PSA and f-PSA. The 4-mercaptobenzoic acid (MBA) on the immunocapture substrate is coupled to the t-PSA antibody through the carboxyl group, and the combination of t-PSA induces the Raman frequency shifts of MBA. The immunocolloidal gold attached with f-PSA antibodies selectively capture the f-PSA that immobilized on the MBA-modified SERS substrates, allowing for f-PSA quantification according to the SERS intensities of the 5, 5'-Dithiobis (succinimidyl-2-nitrobenzoate) (DSNB) probe. The results show that f-PSA and t-PSA have good linear response in the concentration scale of 0.1-20 ng/mL, and 1-200 ng/mL, respectively. The biosensor combines Raman frequency shifts and intensities, which greatly simplifies traditional procedures for f-PSA% detection. All the results demonstrated the great potential of the proposed biosensor in highly reproducible and accurate diagnosis of prostate cancers.