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.

Diabetic nephropathy(DN)is a serious complication of diabetes mellitus and a leading cause of end-stage renal diseases.In DN patients,key pathological mechanisms include proteinuria,glomerulo-sclerosis,and fibrosis,largely driven by poor glycemic control and oxidative stress caused by prolonged hyperglycemia.This stress damages renal podocytes and triggers inflammatory mesenchymal infiltration of renal tubular cells,exacerbating the progression of proteinuria and fibrosis.Human umbilical cord-de-rived mesenchymal stem cells(hUC-MSCs)offer promising potential for treating DN due to their strong anti-oxidative properties.In this study,we developed a DN mouse model and treated the mouse via tail vein injections of hUC-MSCs(1×106 cells/mouse).The results indicated that hUC-MSCs significantly lowered fasting blood glucose levels(22.5±3.0 vs 14.7±1.1,P＜0.01)and improved glucose toler-ance,as shown by intraperitoneal glucose tolerance test(IPGTT)results(P＜0.05).Additionally,the renal function improved in hUC-MSCs-treated mice,with marked reductions in oxidative stress markers,including blood urea nitrogen(BUN),urinary creatinine(Ucr),urinary protein(PRO),superoxide dismutase(SOD),and malondialdehyde(MDA)(P＜0.05).Histological analyses through hematoxy-lin-eosin(H&E),Periodic Acid-Schiff(PAS),and Sirius red staining demonstrated alleviation of glo-merular mesangial hyperplasia,glomerular hypertrophy,and tubular inflammation.Furthermore,hUC-MSCs treatment downregulated the expression of oxidative stress-related proteins,such as NADPH oxi-dase 4(NOX4)and thioredoxin-interacting protein(TXNIP),and reduced reactive oxygen species(ROS)production(P＜0.05).Meanwhile,human renal cortical proximal tubule epithelial cells(HK-2 cells)were selected for validation in vitro experiments using high glucose treatment followed by super-natants of hUC-MSCs(MSC-CM),and Western blotting showed that the expression of both NOX4 and TXNIP was inhibited(P＜0.05)and ROS expression was reduced.In conclusion,hUC-MSC treatment effectively lowered blood glucose levels and improved renal function in DN mice,likely through the sup-pression of NOX4 expression and TXNIP-mediated oxidative stress.

Objective To investigate the changing antibiotic resistance profiles of E.coli isolated from patients in the 52 hospitals participating in the CHINET program from 2015 to 2021.Methods Antimicrobial susceptibility was tested for clinical isolates of E.coli according to the unified protocol of CHINET program.WHONET 5.6 and SPSS 20.0 software were used for data analysis.Results Atotal of 289 760 nonduplicate clinical strains ofE.coli were isolated from 2015 to 2021,mainly from urine samples(44.7±3.2)％.The proportion of E.coli strains isolated from urine samples was higher in females than in males(59.0％vs 29.5％).The proportion of E.coli strains isolated from respiratory tract and cerebrospinal fluid samples was significantly higher in children than in adults(16.7％vs 7.8％,0.8％vs 0.1％,both P＜0.05).The isolates from internal medicine department accounted for the largest proportion(28.9±2.8)％with an increasing trend over years.Overall,the prevalence of ESBLs-producing E.coli and carbapenem resistant E.coli(CREco)was 55.9％and 1.8％,respectively during the 7-year period.The prevalence of ESBLs-producing E.coli was the highest in tertiary hospitals each year from 2015 to 2021 compared to secondary hospitals.The prevalence of CREco was higher in children's hospitals compared to secondary and tertiary hospitals each year from 2015 to 2021.The prevalence of ESBLs-producing E.coli in tertiary hospitals and children's hospitals and the prevalence of CREco in children's hospitals showed a decreasing trend over the 7-year period.The prevalence of CREco in secondary and tertiary hospitals increased slowly.Antibiotic resistance rates changed slowly from 2015 to 2021.Carbapenem drugs(imipenem,meropenem)were the most active drugs amongβ-lactams against E.coli(resistance rate≤2.1％).The resistance rates of E.coli to β-lactam/β-lactam inhibitor combinations(piperacillin-tazobactam,cefoperazone-sulbactam),aminoglycosides(amikacin),nitrofurantoin and fosfomycin(for urinary isolates only)were all less than 10％.The resistance rate of E.coli strains to antibiotics varied with the level of hospitals and the departments where the strains were isolated,especially for cefazolin and ciprofloxacin,to which the resistance rate of E.coli strains from children in non-ICU departments was significantly lower than that of the strains isolated from other departments(P＜0.05).The E.coli isolates from ICU showed higher resistance rate to most antimicrobial agents tested(excluding tigecycline)than the strains isolated from other departments.The E.coli strains isolated from tertiary hospitals showed higher resistance rates to the antimicrobial agents tested(excluding tigecycline,polymyxin B,cefepime and carbapenems)than the strains from secondary hospitals and children's hospitals.Conclusions E.coli is an important pathogen causing clinical infection.More than half of the clinical isolates produced ESBL.The prevalence of CREco is increasing in secondary and tertiary hospitals over the 7-year period even though the overall prevalence is still low.This is an issue of concern.

Purpose To compare the clinical and cardiovascular magnetic resonance(CMR)characteristics of hypertrophic cardiomyopathy(HCM)with and without diabetes mellitus,and to investigate the effect of diabetes mellitus on left ventricular remodeling in HCM patients.Materials and Methods A total of 96 HCM patients diagnosed with CMR in Chengdu Fifth People's Hospital from January 2021 to March 2023,retrospectively.They were divided into HCM with diabetes mellitus group(n=22)and HCM without diabetes mellitus group(n=74)according to whether they had diabetes mellitus.The clinical baseline and CMR data of HCM patients were analyzed,and left ventricular function and strain were compared between the two groups.Results The HCM group with diabetes mellitus was observed to have a significantly higher left ventricular mass index compared to HCM without diabetes mellitus group[(1.33±0.50)g/ml vs.(1.10±0.23)g/ml;t=3.002,P=0.003],along with a significantly lower left ventricular global circumferential strain[-19.69(-21.82,-17.06)%vs.-16.33(-20.84,-13.86)%;Z=543.000,P<0.05]and global longitudinal strain[(-8.69±3.94)%vs.(-10.74±3.62)%;t=2.227,P<0.05)].Furthermore,diabetes mellitus was identified as an independent factor associated with an increased left ventricular mass index(β=0.330,P<0.05)by multivariate linear regression analysis.Conclusion In patients with HCM,diabetes mellitus is independently associated with left ventricular remodeling.The global circumferential strain and longitudinal strain of HCM patients with diabetes mellitus decreases significantly,suggesting that diabetes mellitus has a significant impact on left ventricular strain in HCM patients.

To understand the background of Escherichia coli(E.coli)carried by artificially bred sika deer and the biological characteristics of the isolated strains,such as drug resistance and pathogenicity,in April 2024,we collected 184 fresh deer fecal samples from four deer farms in Luxiang Township,Shuangyang District,Changchun City,Jilin Province,for isolation and cultivation of E.coli.The isolates were tested for drug resistance and biochemical identification with a BD PhoenixTM-100 Automated Microbiology System.The virulence genes were detected with PCR,and the strains were molecularly typed with ERIC-PCR.A total of 165 E.coli strains were isolated from 184 samples of deer feces,with an isolation rate of 89.67%.Twenty strains had a drug resistance phenotype,and the drug resistance rate was 12.12%;these strains included 15 strains of multi-drug resistant bacteria and 11 strains of ESBL-producing bacteria.Virulence gene detection indicated that the sika deer isolates carried multiple diarrhea-associated virulence genes,such as EAST-1(12.12%),eae(1.21%),stx1(7.88%),stx2(7.27%),and STa(1.82%).ERIC-PCR demonstrated that the isolates showed high polymorphism.The ESBL-producing E.coli carried by sika deer are likely to spread drug resistance in the community and livestock population.Some isolates carried multiple diarrhea-associated virulence genes,thus posing a human transmission risk.Therefore,monitoring of drug resistance and virulence genes must be strengthened,and antibiotics must be used reasonably during the breeding process to avoid excessive use and misuse.

AIM To study the chemical constituents from Dictamni Cortex.METHODS The 70％ethanol extract from Dictamni Cortex was isolated and purified by HP-20 macroporous resin,silica gel,MCI,ODS and preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Thirty-three compounds were isolated and identified as rutin(1),apigenin(2),catechin(3),hesperetin(4),leonuriside A(5),androsin(6),2-methoxy-4-acetylphenol-O-α-rhamnopyranosyl-(1"-6')-β-glucopyranoside(7),vanillic acid(8),gallic acid(9),4-hydroxybenzoic acid(10),benzoic acid(11),involcranoside B(12),benzyl β-D-glucopyranoside(13),bphenylethyl-rutinoside(14),1-bromonaphthalene(15),cimifugin(16),9(S),12(S),13(S)-trihydroxyoctadeca-10(E),15(Z)-dienoic acid(17),methyl-9,12,13-trihydroxyoctadeca-10,15-dienoate(18),7,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid(19),vernolic acid(20),9,10(erythro)-dihydroxy-11 E-octadecadienoic acid methyl ester(21),(7Z,9E,13Z)-11-hydroxyhexadeca-7,9,13-trienoic acid(22),(7Z,10Z,14E,16Z,19Z)-13-hydroxydocosa-7,10,14,16,19-pentaenoic acid(23),(9E)-8,11,12-trihydroxyoctadecenoic acid methyl ester(24),n-hexanol-O-rutinoside(25),hexyl β-sophoroside(26),3-pentyl 6'-(3-hydroxy-3-methylglutaryl)-β-D-glucopyranoside(27),3-methylbut-3-enyl-6-O-β-D-glucopyranosyl-β-D-glucopyranoside(28),3-methyl-but-2-en-1-yl β-D-glucopyranoside(29),3-methylbutan-1-ol-β-D-glucopyranoside(30),pregnenolone(31),2-butoxytetrahydrofuran(32),psydrin(33).CONCLUSION Compounds 2-4,8-13,15-16,25-28 and 32-33 are isolated from Rutaceae family for the first time.

In the past 6 years, significant breakthroughs have been achieved in the treatment of heart failure (HF), especially in drug therapy. The classification of chronic HF and the treatment methods for HF and its complications are also constantly being updated. In order to apply these results to the diagnosis and treatment of patients with HF in China and further improve the level of diagnosis and treatment of HF in China, the HF Group of Chinese Society of Cardiology, Chinese Medical Association, Chinese College of Cardiovascular Physician, Chinese HF Association of Chinese Medical Doctor Association, and Editorial Board of Chinese Journal of Cardiology have organized an expert group and update the consensus and evidence-based treatment methods in the field of HF based on the latest clinical research findings at home and abroad, combined with the national conditions and clinical practice in China, and referring to the latest foreign HF guidelines while maintaining the basic framework of the 2018 Chinese Guidelines for Diagnosis and Treatment of HF.

Atrial fibrillation (AF) is the most common arrhythmia in clinical practice. It has a high prevalence and poor prognosis. The application of antiarrhythmic drugs and even surgery cannot completely treat the disease, and there are many sequelae. AF can be classified into the category of "palpitation" in Chinese medicine according to its symptoms. Acupuncture has a significant effect on AF. The authors find that an important mechanism of acupuncture in AF treatment is to regulate the cardiac vagus nerve. Therefore, this article intends to review the distribution and function of vagus nerve in the heart, the application and the regulatroy effect for the treatment of AF.
Atrial Fibrillation/physiopathology* ; Humans ; Acupuncture Therapy ; Vagus Nerve/physiology* ; Animals

OBJECTIVE: Lipid oxidation is involved in the pathogenesis of atherosclerosis and may be contribute to the development of Ischemic stroke (IS). However, the lipid profiles associated with IS have been poorly studied. We conducted a pilot study to identify potential IS-related lipid molecules and pathways using lipidomic profiling. METHODS: Serum lipidomic profiling was performed using LC-MS in 20 patients with IS and 20 age- and sex-matched healthy controls. Univariate and multivariate analyses were simultaneously performed to identify the differential lipids. Multiple testing was controlled for using a false discovery rate (FDR) approach. Enrichment analysis was performed using MetaboAnalyst software. RESULTS: Based on the 294 lipids assayed, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) models were used to distinguish patients with IS from healthy controls. Fifty-six differential lipids were identified with an FDR-adjusted P less than 0.05 and variable influences in projection (VIP) greater than 1.0. These lipids were significantly enriched in glycerophospholipid metabolism (FDR-adjusted P = 0.009, impact score = 0.216). CONCLUSIONS Serum lipid profiles differed significantly between patients with IS and healthy controls. Thus, glycerophospholipid metabolism may be involved in the development of IS. These results provide initial evidence that lipid molecules and their related metabolites may serve as new biomarkers and potential therapeutic targets for IS.
Humans ; Pilot Projects ; Lipidomics ; Male ; Female ; Biomarkers/blood* ; Middle Aged ; Ischemic Stroke/blood* ; Aged ; China ; Lipids/blood* ; Adult ; Case-Control Studies ; East Asian People