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.

Optical imaging is highly valued for its superior temporal and spatial resolution. This is particularly important in near-infrared II (NIR-II, 1 000-3 000 nm) imaging, which offers advantages such as reduced tissue absorption, minimal scattering, and low autofluorescence. These characteristics make NIR-II imaging especially suitable for deep tissue visualization, where high contrast and minimal background interference are critical for accurate diagnosis and monitoring. Currently, inorganic fluorescent probes—such as carbon nanotubes, rare earth nanoparticles, and quantum dots—offer high brightness and stability. However, they are hindered by ambiguous structures, larger sizes, and potential accumulation toxicity in vivo. In contrast, organic fluorescent probes, including small molecules and polymers, demonstrate higher biocompatibility but are limited by shorter emission wavelengths, lower quantum yields, and reduced stability. Recently, gold clusters have emerged as a promising class of nanomaterials with potential applications in biocatalysis, fluorescence sensing, biological imaging, and more. Water-soluble gold clusters are particularly attractive as fluorescent probes due to their remarkable optical properties, including strong photoluminescence, large Stokes shifts, and excellent photostability. Furthermore, their outstanding biocompatibility—attributed to good aqueous stability, ultra-small hydrodynamic size, and high renal clearance efficiency—makes them especially suitable for biomedical applications. Gold clusters hold significant potential for NIR-II fluorescence imaging. Atomic-precision gold clusters, typically composed of tens to hundreds of gold atoms and measuring only a few nanometers in diameter, possess well-defined three-dimensional structures and clear spatial coordination. This atomic-level precision enables fine-tuned structural regulation, further enhancing their fluorescence properties. Variations in cluster size, surface ligands, and alloying elements can result in distinct physicochemical characteristics. The incorporation of different atoms can modulate the atomic and electronic structures of gold clusters, while diverse ligands can influence surface polarity and steric hindrance. As such, strategies like alloying and ligand engineering are effective in enhancing both fluorescence and catalytic performance, thereby meeting a broader range of clinical needs. In recent years, gold clusters have attracted growing attention in the biomedical field. Their application in NIR-II imaging has led to significant progress in vascular, organ, and tumor imaging. The resulting high-resolution, high signal-to-noise imaging provides powerful tools for clinical diagnostics. Moreover, biologically active gold clusters can aid in drug delivery and disease diagnosis and treatment, offering new opportunities for clinical therapeutics. Despite the notable achievements in fundamental research and clinical translation, further studies are required to address challenges related to the standardized synthesis and complex metabolic behavior of gold clusters. Resolving these issues will help accelerate their clinical adoption and broaden their biomedical applications.

Localized scleroderma (LoS) is a chronic autoimmune skin disorder characterized by fibrosis and hardening of the skin and subcutaneous tissues, potentially affecting deeper tissues and other organs. Currently, China lacks evidence-based guidelines for the diagnosis and management of LoS, which poses a challenge to disease recognition and leads to significant variation in treatment strategies across medical centers. To address this problem, we plan to establish a nationwide, multidisciplinary expert team through the Chinese Society of Plastic Surgery to initiate the development of the Chinese Guideline for the Diagnosis and Management of Localized Scleroderma. The guideline aims to standardize the diagnosis and treatment procedures for LoS, improve clinical diagnostic and therapeutic capabilities, and enhance patient outcomes through early identification and intervention. The guideline will adhere to international standards, with recommendations formed through evidence evaluation and multiple rounds of expert discussions. Although evidence-based data on LoS specific to the Chinese population is currently limited, and no unified gold standard for disease assessment exists, the clinical expertise of multidisciplinary specialists will play a pivotal role in the guideline's formulation. We anticipate that this guideline will provide LoS patients with higher-quality medical care and enhance patient awareness and engagement, which may ultimately help reduce social and healthcare burden. This proposal outlines the background, methodology, and anticipated value of the guideline, with the hope of providing guidance for its rigorous development and successful publication.

Localized scleroderma (LoS) is a chronic autoimmune skin disorder characterized by fibrosis and hardening of the skin and subcutaneous tissues, potentially affecting deeper tissues and other organs. Currently, China lacks evidence-based guidelines for the diagnosis and management of LoS, which poses a challenge to disease recognition and leads to significant variation in treatment strategies across medical centers. To address this problem, we plan to establish a nationwide, multidisciplinary expert team through the Chinese Society of Plastic Surgery to initiate the development of the Chinese Guideline for the Diagnosis and Management of Localized Scleroderma. The guideline aims to standardize the diagnosis and treatment procedures for LoS, improve clinical diagnostic and therapeutic capabilities, and enhance patient outcomes through early identification and intervention. The guideline will adhere to international standards, with recommendations formed through evidence evaluation and multiple rounds of expert discussions. Although evidence-based data on LoS specific to the Chinese population is currently limited, and no unified gold standard for disease assessment exists, the clinical expertise of multidisciplinary specialists will play a pivotal role in the guideline's formulation. We anticipate that this guideline will provide LoS patients with higher-quality medical care and enhance patient awareness and engagement, which may ultimately help reduce social and healthcare burden. This proposal outlines the background, methodology, and anticipated value of the guideline, with the hope of providing guidance for its rigorous development and successful publication.

Tranexamic acid is widely used in joint orthopedic surgery.At the same time,it has high safety and few adverse drug reactions.It can effectively improve intraoperative bleeding and promote early functional recovery of patients.This article reviews the mode of administration,safe dose,administration time and adverse drug reactions of tranexamic acid in the perioperative period of joint replacement and arthroscopic surgery,in order to provide reference for the clinical application of tranexamic acid.

MADS-box protein family are important transcriptional regulatory factors in plant growth and development. The AGAMOUS 12 (AGL12) subfamily is believed to play an important regulatory role in the process of plant flowering transition. To explore the potential mechanism of AGL12 subfamily involved in regulating the flower development of Lonicera macranthoides, quantitative real-time polymerase chain reaction (qRT-PCR), prokaryotic expression and yeast two-hybrid techniques were used to analyze the expression pattern, protein expression, and protein-protein interaction pattern of LmAGL12 based on transcriptome data. The results showed that the LmAGL12 gene contains a 603 bp open reading frame (ORF), encoding 200 amino acids, and the encoded protein was stable and hydrophilic without a transmembrane region and signal peptide. Through homologous sequence alignment and phylogenetic analysis, it was confirmed that LmAGL12 protein belongs to the MADS-box protein family and is closely related to the AGL12 protein of Heracleum sosnowskyi and Daucus carota subsp. sativus. The LmAGL12 gene was cloned into prokaryotic expression vector pET-28a and the recombinant constructs were transformed into Escherichia coli BL21 (DE3), which inducing the target protein successfully. The yeast two-hybrid results showed that LmAGL12 protein interacts with LmSVP protein, LmSOC1 protein and LmAP1 protein, respectively. The qRT-PCR results showed that LmAGL12 gene were differentially expressed in different development stages of flower bud, stem, and leaves of 'Longhua' and 'Baiyun' in L. macranthoides. The LmAGL12 gene showed the highest expression level in the middle stage of the 'Longhua' floral bud; For the 'Baiyun' variety, the relative expression level of LmAGL12 gene is the highest in the stem. This study cloned the LmAGL12 gene in L. macranthoides and analyzed it expression for the first time, enriching the research on flower organ development and providing a research basis for further exploring the molecular mechanisms of long bud stage and non-unfolded corolla in L. macranthoides, as well as for variety improvement.

Objective:To assess the efficacy and safety of overlapping braided carotid artery stent (Wallstent) implantation in large extracranial internal carotid artery aneurysms (15 mm≤diameter<25 mm) and giant ones (diameter≥25 mm).Methods:A retrospective study was performed; the clinical data of 23 patients with large or giant extracranial internal carotid artery aneurysms accepted overlapping Wallstent stent implantation in Department of Interventional Radiology, First Affiliated Hospital of Zhengzhou University from August 2015 to June 2023 were collected. Immediately after implantation, DSA was used to evaluate the retention of contrast agent within the aneurysms and high-resolution C-arm CT (HR-CBCT) was used to detect the apposition between the two stents and between the stents and inner wall of the blood vessel. Perioperative complications were recorded. Clinical follow-up was performed bi-monthly via outpatient visits or telephone, and modified Rankin scale (mRS) was used to assess the prognoses (mRS scores of 0-2 as good prognosis) at the last follow-up; aneurysm occlusion was evaluated in a 6-month follow-up by DSA and in-stent restenosis in a final imaging follow-up by DSA or CTA according to the OKM grading. Results:Twenty-two patients had successful overlapping implantation of 2 Wallstent stents; blood flow was restricted in one patient due to carotid artery dissection at the distal end of the aneurysm during stent implantation and restored after a Neuroform EZ stent and 2 Wallstent stent implantation from the distal-proximal lesion; technical success rate of 95.7% (22/23) was obtained. DSA immediately after implantation showed obvious contrast medium retention in all aneurysms. HR-CBCT indicated good stent apposition in 21 patients and mild incomplete stent apposition in 2. Clinical follow-up was finished in 23 patients, ranged 6-31 months (mean 11.5±6.3 months); all patients had good prognosis at the last follow-up. Imaging follow-up, including at least once DSA, was conducted for all patients, with intervals ranging from 6 to 15 months (mean 10.4±3.4 months); DSA 6 months after implantation showed complete aneurysm occlusion in 19 patients (OKM grading D) and a bit of residual contrast in 4 patients (OKM grading C); final imaging follow-up (DSA in 2 and CTA in 21) revealed in-stent stenosis in 2 patients (stenosis rates of 51% and 87%) with obvious improved stenosis after balloon angioplasty and patent stents in 21 patients without evidence of aneurysm opacification.Conclusion:Overlapping braided carotid artery stent (Wallstent) implantation is an effective and safe approach for managing large or giant extracranial carotid artery aneurysms.

Mitral valve prolapse is one of the common causes of mitral regurgitation.Mitral valve prolapse complicated with leaflet cleft is rare in clinical practice,which most often undergo surgical mitral valve repair or mitral valve replacement.We report a case of mitral valve prolapse with posterior leaflet cleft treated by transcatheter edge-to-edge mitral valve repair,in order to provide a model for similar cases.

Objective:To explore the potential causal relationship between gut microbiota and teratozoospermia.Methods:We searched the database of Genome-Wide Association Study(GWAS)for gut microbiota-and teratozoospermia-related data.We used gut microbiota as an exposure factor,determined the instrumental variables according to the GWAS data on 18 340 participants released by the MiBioGen Alliance,and derived the outcome variables from the European data on teratozoospermia,with a sample size of 85 716,including 915 cases and 209 006 controls.Using inverse-variance weighting(IVW),MR-Egger regression and the weighted median estimator(WME),we performed two-sample Mendelian randomization(MR)analysis on the retrieved data,and estimated the causal relationship between gut microbiota and teratozoospermia based on the β value.Results:Two-sample MR analysis indicated that the class Erysipelotrichia,family Erysipelotrichaceae,family Streptococcaceae,genus Coprococcusl,genus Ruminococcaceae UCG009,genus Streptococcus,order Clostridialesm and order Erysipelotrichales were causally related with the increased risk,while the family Porphyromonadaceae with the decreased risk of teratozoospermia.Conclusion:The class Erysipelotrichia,family Erysipe-lotrichaceae,family Streptococcaceae,genus Coprococcusl,genus Ruminococcaceae UCG009,genus Streptococcus,order Clostridia-lesm and order Erysipelotrichales are one of the causes of teratozoospermia,related to the increased risk of the condition,while the family Porphyromonadaceae has a protective effect on sperm morphology,reducing the risk of teratozoospermia.

Objective To explore the clinical characteristics of patients with alcoholic liver disease(ALD)of various traditional Chinese medicine(TCM)syndrome types.Methods A retrospective analysis was conducted in 129 patients with alcoholic liver disease who met the inclusion and exclusion criteria in Foshan Hospital of Traditional Chinese Medicine from 2018 to 2022.The general data of the patients as well as their TCM syndrome types and clinical information of liver and kidney function,blood lipid,liver transient elastography during the hospital visit were collected.The distribution of TCM syndrome types in ALD patients was analyzed,and the clinical characteristics of the ALD patients with various TCM syndrome types were explored.Results(1)Of the 129 patients,128(99.22%)were male and only one(0.78%)was female,the average age was(48.71±11.50)years old,and the average body mass index(BMI)was(23.82±3.98)kg·m-2.(2)Damp-heat accumulation syndrome was most common syndrome type in ALD patients,with a total of 70 cases(54.26%),and then came liver depression and spleen deficiency syndrome(24 cases,18.60%),internal obstruction of phlegm-damp syndrome(22 cases,17.05%),liver-kidney sufficiency syndrome(7 cases,5.43%),phlegm interweaved with blood stasis syndrome(3 cases,2.33%),and internal accumulation of blood stasis syndrome(3 cases,2.33%).(3)The analysis of clinical characteristics by non-parametric rank sum test showed that there were no statistically significant differences in BMI,alcohol consumption,aspartate aminotransferase(AST),gamma-glutamyltransferase(GGT),total bilirubin(TBIL),alkaline phosphatase(ALP),triglyceride(TG),liver stiffness measurement(LSM),and controlled attenuation parameter(CAP)which reflects the fat content of liver in ALD patients with various TCM syndrome types(P＜0.05 or P＜0.01).The prominent features were as follows:patients with the 4 types of liver depression and spleen deficiency,internal obstruction of phlegm-damp,phlegm interweaved with blood stasis,and internal accumulation of blood stasis had a BMI exceeding the standard(＞24 kg·m-2),whereas patients with damp-heat accumulation syndrome and liver-kidney deficiency syndrome,which accounted for 54.26%of the sample size,had a BMI within the normal range(23.03 kg·m-2 and 21.42 kg·m-2,respectively),and the BMI of these two types differed from that(26.44 kg·m-2)of the internal obstruction of phlegm-damp syndrome(P＜0.01),suggesting that more than half of the ALD patients had the normal BMI;moreover,the patients with internal obstruction of phlegm-damp also had the highest values of serum TG(2.69 mmol/L)and CAP(292 db/m)except for the highest BMI,indicating that patients with internal obstruction of phlegm-damp syndrome had a more serious degree of obesity and hepatic fat infiltration than those with other syndrome types;the levels of AST and GGT,which separately reflect the chronic inflammatory injury of liver and bile duct cell injury,were significantly increased in the patients with damp-heat accumulation syndrome and liver-kidney deficiency syndrome,and the LSM value of these two types of patients was also the highest in all of the syndrome types,the differences being all statistically significant(P＜0.05 or P＜0.01).Conclusion Damp-heat accumulation syndrome is the main TCM syndrome type of ALD patients,the degree of fatty infiltration of the liver and overweight of ALD patients are not corresponded to the severity of illness,and there are some differences in the clinical indicators of ALD patients with various TCM syndrome types.However,with cross reference to the data of the four diagnostic examinations of TCM and the clinical indicators,the accuracy of the TCM diagnosis of ALD is expected to be increased.