Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

Chemiluminescence,as a self-luminous phenomenon that does not require light,heat,acoustic,electric and magnetic excitation,has been widely used in the fields of analytical chemistry,cold light source and bio-imaging because of its advantages including high sensitivity,wide linear range,simple equipment and fast detection speed compared with other analytical techniques.Carbon dots(CDs)are a class of nanomaterials with excellent photoluminescence properties and high biocompatibility.CDs are stable,easy to prepare and abundant in types,and researchers have introduced many types of CDs into different chemiluminescence systems.In this paper,the applications of CDs in common chemiluminescence systems and the possible mechanisms of action were discussed,and the research progresses on the application of CDs in different chemiluminescence detection fields in recent years were summarized.Finally,the development trend of CDs in chemiluminescence was analyzed.

Poly(lactic-co-glycolide)(PLGA)is a key excipient in long-acting sustained-release preparations,and its degradation properties directly affect the drug release behavior.In this study,PLGA microspheres were prepared by microfluidic techniques,and the morphology changes of the microspheres were observed by scanning electron microscopy(SEM).In alkaline environment,due to the accelerated hydrolysis of ester bonds,the surface of the microspheres was rapidly dissolved and eroded,and the degradation rate was significantly higher than that in acidic environment.High temperature accelerated the degradation of PLGA microspheres.Under neutral and alkaline conditions,the microspheres showed aggregation and adhesion.Under acidic conditions,the microspheres gradually decomposed into irregular fragments.The high ionic strength further promoted the surface corrosion of the microspheres,especially under extreme pH conditions.Simultaneously,PLGA microspheres encapsulating coumarin were prepared to simulate the microsphere formulation.The release rate of coumarin after degradation of the microspheres under different conditions was observed by measuring the absorbance with ultraviolet-visible spectrophotometry.The results were consistent with those of the blank microspheres.This study revealed that the degradation of PLGA microspheres was significantly pH-dependent,temperature sensitive and ion strength responsive.These findings not only helped to understand and optimize the long-term stability and controlled release performance of drug-carrying microspheres,but also provided a theoretical basis for further improvement of PLGA-based drug carrier design.

Introducing fingerprint level 3 features(especially sweat pores)in fingerprint recognition can significantly improve the value of fingerprints.However,conventional fingerprint visualization methods suffer from issues such as poor stability and reproducibility,insufficient resolution,and feature masking in detecting level 3 features.Electrospun membrane has unique advantages in latent fingerprint(LFP)detection due to its excellent adsorption performance and high specific surface area,and thus its application potential in LFP visualization urgently need to be explored.A novel pore visualization method based on core-shell structured PAN-Flu/PVP composite nanofibrous membrane was proposed in this work.Specifically,the PAN-Flu/PVP composite nanofibrous membrane was prepared via coaxial electrospinning technology,with polyacrylonitrile(PAN)loaded with fluorescein(Flu)as the core and polyvinylpyrrolidone(PVP)as the shell.The experimental results showed that the prepared PAN Flu/PVP composite nanofibrous membrane had a porous structure and excellent adsorption performance.Based on the water solubility of the outer shell PVP and the water induced fluorescence enhancement effect of the core Flu,high-resolution visualization of sweat pores could be achieved within 2 s.The optimization experiment showed that the best quality of sweat latent fingerprints was obtained when the Flu content was 4 mg/mL,the spinning time was 1 h,and the sweating time was 2 min.Through repeated fingerprinting and live fingerprint comparison experiment,the strong stability and high reproducibility of the as-produced membrane in displaying fingerprint sweat pores were finally verified.In summary,the development method could quickly,stably and accurately extract the spatial distribution and activity level of fingerprint sweat pores,which was of great significance for improving the utilization and value of fingerprints.

Due to the poor ionization efficiency and the weak mass spectrometry(MS)intensity of weakly polar substances,direct analysis using the traditional electrospray ionization mass spectrometry(ESI-MS)is a big challenge.In this study,a novel rapid on-site detection method of four prohibited sex hormones in cosmetics was proposed using online derivatization strategy coupled with a miniature mass spectrometer.The target substances in the samples were extracted by a custom-made polyaniline/multi-walled carbon nanotube solid-phase microextraction(SPME)probe.The stirring speed was 200 r/min,the extraction temperature was 40℃,and the extraction time was 2 min.A pulled dual-channel θ borosilicate glass capillary emitter was used as the nano-ESI ion source.The SPME probe was inserted into the channel containing methanol in theθborosilicate glass capillary.When the spray voltage was applied,the four sex hormones were desorbed and formed spray microdroplets,which then collided with the hydroxylamine microdroplets generated from the other channel.The microdroplets of reaction product entered into the miniature mass spectrometer for direct analysis.The limits of detection(LOD)and limits of quantification(LOQ)for the four sex hormones were 10-20 ng/mL and 20-50 ng/mL,respectively.The recoveries were from 84.6%to 107.8%with the relative standard deviations(RSD)from 4.1%to 11.6%.Compared to detection without derivatization,the MS signals of the four target substances were increased by 3 to 15 times.This method was simple,rapid,highly efficient and sensitive,and suitable for on-site rapid analysis of weakly polar sex hormones in cosmetics.

Objectives:To evaluate the predictive value of Tabular Prior-data Fitted Network(TabPFN) for short-term outcome in patients with spontaneous intracerebral hemorrhage (sICH), and compared with the Extreme Gradient Boosting (XGboost) model and traditional logistic regression (LR) model. Methods:Patients with sICH admitted to the Department of Neurology, Hefei Second People's Hospital from January 2018 to March 2024 were included retrospectively. The demographic and baseline data were collected. At 3 months after onset, the modified Rankin Scale score was used to determine the outcome, 0-2 was defined as good outcome and >2 was defined as poor outcome. All enrolled patients were randomly divided into a training set and a testing set at a ratio of 7:3. Feature selection was performed using recursive feature elimination (RFE) method, and then the selected feature variables were included into TabPFN, XGboost, and LR models for training and testing. The area under the curve (AUC) of receiver operating characteristic (ROC) curve was used to evaluate the predictive ability of the models. Shapley additive explanations (SHAP) method was used for model interpretation.Results:A total of 547 patients with sICH were enrolled, including 367 males (67.1%), with a median age of 65 (interquartile range, 54-76) years. Two hundred twenty-six patients (41.3%) had poor outcome. Age, baseline blood pressure (systolic blood pressure, diastolic blood pressure), baseline laboratory tests (white blood cell count, red blood cell count, platelet count, neutrophil count, hemoglobin, fasting blood glucose, creatinine, uric acid, urea nitrogen, alanine aminotransferase, aspartate aminotransferase), hematoma rupture into the ventricle, island sign, baseline hematoma volume, and baseline National Institutes of Health Stroke Scale (NIHSS) score were selected as characteristic variables using RFE method. ROC curve analysis showed that the ROC AUC for TabPFN, Xgboost, and LR models predicting poor short-term outcome in the testing set were 0.918 (95% confidence interval [ CI] 0.870-0.966], 0.883 (95% CI 0.826-0.940), and 0.905 (95% CI 0.854-0.957), respectively. SHAP analysis showed that the top four important variables in the TabPFN model were baseline NIHSS score, baseline hematoma volume, baseline aspartate aminotransferase, and age. Conclusions:The TabPFN model is superior to the LR model and the XGBoost model in predicting poor outcome in patients with sICH. In the TabPFN model, baseline NIHSS score, baseline hematoma volume, aspartate aminotransferase, and age are the most important predictors of poor outcome in patients with sICH.Objectives To evaluate the predictive value of Tabular Prior-data Fitted Network(TabPFN) for short-term outcome in patients with spontaneous intracerebral hemorrhage (sICH), and compared with the Extreme Gradient Boosting (XGboost) model and traditional logistic regression (LR) model. Methods Patients with sICH admitted to the Department of Neurology, Hefei Second People's Hospital from January 2018 to March 2024 were included retrospectively. The demographic and baseline data were collected. At 3 months after onset, the modified Rankin Scale score was used to determine the outcome, 0-2 was defined as good outcome and >2 was defined as poor outcome. All enrolled patients were randomly divided into a training set and a testing set at a ratio of 7:3. Feature selection was performed using recursive feature elimination (RFE) method, and then the selected feature variables were included into TabPFN, XGboost, and LR models for training and testing. The area under the curve (AUC) of receiver operating characteristic (ROC) curve was used to evaluate the predictive ability of the models. Shapley additive explanations (SHAP) method was used for model interpretation. Results A total of 547 patients with sICH were enrolled, including 367 males (67.1%), with a median age of 65 (interquartile range, 54-76) years. Two hundred twenty-six patients (41.3%) had poor outcome. Age, baseline blood pressure (systolic blood pressure, diastolic blood pressure), baseline laboratory tests (white blood cell count, red blood cell count, platelet count, neutrophil count, hemoglobin, fasting blood glucose, creatinine, uric acid, urea nitrogen, alanine aminotransferase, aspartate aminotransferase), hematoma rupture into the ventricle, island sign, baseline hematoma volume, and baseline National Institutes of Health Stroke Scale (NIHSS) score were selected as characteristic variables using RFE method. ROC curve analysis showed that the ROC AUC for TabPFN, Xgboost, and LR models predicting poor short-term outcome in the testing set were 0.918 (95% confidence interval [ CI] 0.870-0.966], 0.883 (95% CI 0.826-0.940), and 0.905 (95% CI 0.854-0.957), respectively. SHAP analysis showed that the top four important variables in the TabPFN model were baseline NIHSS score, baseline hematoma volume, baseline aspartate aminotransferase, and age. Conclusions The TabPFN model is superior to the LR model and the XGBoost model in predicting poor outcome in patients with sICH. In the TabPFN model, baseline NIHSS score, baseline hematoma volume, aspartate aminotransferase, and age are the most important predictors of poor outcome in patients with sICH.

ObjectiveTo characterize the seasonal patterns of influenza in Kunming City, Yunnan Province before, during, and after the COVID-19 pandemic, and provide scientific evidence for optimizing influenza prevention and control strategies. MethodsInfluenza-like illness (ILI) and etiological surveillance data for influenza from the 14^th week of 2010 to the 13^th week of 2024 in Kunming City of Yunnan Province were collected. Harmonic regression models were constructed to analyze the epidemic characteristics and seasonal patterns of influenza before (2010/2011‒2019/2020 influenza seasons), during (2020/2021‒2022/2023 influenza seasons), and after (2023/2024 influenza season) the COVID-19 pandemic. ResultsBefore the COVID-19 pandemic, influenza in Kunming City mainly exhibited an annual cyclic pattern without a significant semi-annual periodicity, peaking from December to February of the next year, with an epidemic duration of 20‒30 weeks. During the pandemic, influenza seasonality shifted, with an increase in semi-annual periodicity and an approximate one month delay in annual peaks. However, after the pandemic, the annual amplitude of influenza increased compared with that before the pandemic, and the epidemic duration extended by about one month. Although the annual peak largely reverted to the pre-pandemic levels, the annual peaks for different influenza subtypes/lineages had not fully recovered. ConclusionInfluenza seasonality in Kunming City underwent substantial alterations following the COVID-19 pandemic and has not yet fully reverted to pre-pandemic levels. Continuous surveillance on different subtypes/lineages of influenza viruses remains essential, and prevention and control strategies should be adjusted and optimized in a timely manner based on current epidemic trends.

Rare tumors pose significant challenges to clinical research, diagnosis, and therapy due to their low incidence, high molecular heterogeneity, and the lack of preclinical models.Conventional patient-derived cell lines and patient-derived tumor xenografts have significant limitations in recapitulating the characteristics of rare tumors. As an emerging in vitro biological model, organoids preserve the molecular characteristics of original tumors and possess high biological relevance, efficient amplification capabilities, and potential for personalized medicine applications. They have currently been applied in the research of various rare tumors. This review summarizes current research on organoid models of rare tumors derived from different tissue origins, and outlines current challenges and future development directions.

AIM To investigate the effect of Fuzheng Hefu Zhiyang Formula(FZHFZY)on psoriasis-like skin lesions and immune regulation in mice.METHODS In the in vivo experiment,30 BALB/c mice were randomly divided into the blank group,the model group,the dexamethasone group(1.5 g/kg of compound dexamethasone acetate cream),and the low-dose(2.5 g/kg)and high-dose(5 g/kg)FZHFZY groups,with six mice in each group.The experiment groups were treated with respective FZHFZY and dexamethasone,and the other groups were given normal saline for 10 consecutive days,during which psoriatic skin lesions were induced with imiquimod cream for 7 consecutive days.The mice had their area and severity of psoriasis assessed by PASI score;their histological changes of skin lesions.observed with Hematoxylin-eosin(HE)staining;their F4/80 ratio of skin lesions observed with immunohistochemical(IHC)staining;their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected by Western blot;and their mRNA expressions of tumor necrosis factor-α(TNF-α),IL-17,IL-23 and IL-1β detected by RT-qPCR.In the in vitro research,the cultured RAW264.7 cells were divided into the blank group,the LPS group,and the FZHFZY groups(1 200,600,300,150 μg/mL).The cells had their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected with Western blot;and their mRNA expressions of IL-6,TNF-α,IL-23 and IL-8 detected by RT-qPCR.RESULTS The in vivo experiment showed that compared to the model group,the FZHFZY groups demonstrated decreased PASI score(P＜0.01);improved epidermal thickening and parakeratosis of skin lesions as revealed by HE staining result and increased expression of F4/80 in IHC staining sections;decreased protein expression ratios of p-P38/P38,p-ERK/Erk and p-P65/P65 in skin(P＜0.05,P＜0.01);and reduced mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β in the skin(P＜0.01).FZHFZY(0～2 400 μg/mL)showed no significant cytotoxicity towards RAW264.7 cells in vitro(P＞0.05).Compared to those of the LPS group,the cells exposed to FZHFZ at concentrations of 1 200 and 600 μg/mL demonstrated decreased protein expression ratios of p-P38/P38,p-ERK/Erk,and p-P65/P65(P＜0.05,P＜0.01);and significantly decreased mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β(P＜0.01).CONCLUSION FZHFZY alleviates imiquimod-induced psoriatic lesions in mice and suppresses inflammatory response in LPS-stimulated RAW264.7 cells by inhibiting P38/Erk/NF-κB signaling pathway.

Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.