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.

Objective To explore the correlation between stroke etiology and clinical and imaging features in patients with acute ischemic stroke(AIS)due to large vessel occlusion treated by intravascular thrombectomy.Methods A total of 213 patients with AIS and endovascular embolectomy in our hospital from Oct.2016 to Jun.2018 were enrolled retrospectively.According to the etiological classification criteria of Trial of Org 10172 in Acute Stroke Treatment(TOAST),there were 116 cases of cardioembolism and 97 cases of non-cardioembolism.Multivariate logistic regression analysis was used to screen the clinical and imaging characteristics for identifying cardioembolism and non-cardioembolism.Results Compared with non-cardioembolism AIS,cardioembolism AIS was associated with higher NIHSS scores(adjusted odds ratio[OR]=1.09,95%confidence interval[95%CI]1.01-1.18,P=0.02),atrial fibrillation(adjusted OR=76.46,95%CI 26.75-218.51,P＜0.01),absence of hypertension(adjusted OR=0.32,95%CI 0.12-0.84,P=0.02),antiplatelet drug use(adjusted OR=5.03,95%CI 1.22-20.63,P=0.03),shorter onset-to-puncture time(adjusted OR=0.998,95%CI 0.996-1.000,P=0.04),and presence of hyperdense artery sign(HAS)(adjusted OR=4.45,95%CI 1.47-13.49,P=0.01).Conclusion There are some differences in clinical and imaging characteristics between patients with cardioembolism and non-cardioembolism AIS.The occurrence of HAS suggests a higher probability of cardioembolism in AIS patients.

Aim To investigate the predictive value of lipoprotein(a)[Lp(a)]combined with systemic inflam-matory response index(SIRI)on in-stent restenosis(ISR)after percutaneous coronary intervention(PCI)in patients with coronary heart disease.Methods The clinical data of 770 patients with coronary heart disease who underwent PCI in the Department of Cardiovascular Medicine of the First Affiliated Hospital of Xinjiang Medical University from May 2012 to December 2024 and underwent coronary angiography six months after surgery were collected.According to the imaging re-sults,the patients were divided into ISR group(n=194)and non-ISR group(n=576).Multivariate Logistic regression and random forest model were used to analyze the independent risk factors of ISR.Risk factors included in the analysis were glycated hemoglobin,SIRI,Lp(a),lymphocyte count,apolipoprotein A1(ApoA1)and residual cholesterol.Results The levels of Lp(a)and SIRI in the ISR group were significantly higher than those in the non-ISR group(P＜0.05).ROC curve analysis showed that the area under the curve of the combined indicator of Lp(a)and SIRI was 0.789,which was higher than the 0.652 of the single indicator Lp(a)and 0.778 of SIRI.Conclusion Lp(a)and SIRI are independent risk factors for the occurrence of ISR after PCI,and the combination of Lp(a)and SIRI can better predict the occurrence of ISR.

Objective:To analyze the symptoms,diagnosis and treatment of upper urinary tract calculi patients combined with mild and moderate benign prostatic hyperplasia(BPH)after ureteral stent implantation.Methods:One hundred and six BPH pa-tients who were hospitalized for upper urinary tract calculi and had ureteral stents retained from January 2019 to December 2022 were selected and divided into 2 weeks group and 4 weeks group according to the time of removal of ureteral stents after surgery.Their gener-al clinical data were analyzed and compared.International Prostatic Symptom Scale(IPSS),postoperative ureteral Stent Symptom Questionnaire(USSQ),and incidence of adverse events after ureteral stent removal were recorded before and after removal.Results:The scores of IPSS were significantly increased in all patients,and symptoms in urinary tract had improved significantly after discharge(P＜0.05).Compared with the 2 weeks group,the USSQ score of the 4 weeks group was significantly increased(P＜0.05).And no significant adverse event was observed in the 2 weeks group after the removal of ureteral sten.Conclusion:IPSS score and USSQ score increased significantly during stent implantation in BPH patients with lithiasis.And complications increased sig-nificantly over time.Following thorough clinical assessment,early ureteral stent removal demonstrates both safety and efficacy,repre-senting an optimal therapeutic approach in selected cases.

Objective To evaluate the clinical application value of the half-Fourier acquisition single-shot turbo spin echo(HASTE)sequence based on deep learning(DL)in pancreatic T2WI.Methods Data were collected from 41 patients who un-derwent both BLADE and DL-HASTE sequence scans during pancreatic T2WI at some hospital from February to July 2023.Qualitative assessments were made regarding overall image quality,pancreatic edge sharpness,pancreatic duct edge sharp-ness,pancreatic duct visua-lization(proximal,middle and distal segments)and lesion visibility of BLADE-sequence and DL-HASTE-sequence images.Quantitative assessments were carried out in terms of scan time,signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR).Statistical analyses were performed using SPSS 24.0.Results DL-HASTE sequence behaved significantly better than BLADE in pancreatic duct edge sharpness,pancreatic duct visualization,lesion visibility,scan time and CNR,while worse in pane-reatic edge sharpness(all P＜0.05).There were significant differences between DL-HASTE and BLADE sequences in overall image quality and SNR(all P＞0.05).Conclusion The DL-HASTE sequence maintains image quality while significantly shor-tening scan time,making it suitable for patients with irregular respiratory rates.[Chinese Medical Equipment Journal,2025,46(3):59-63]

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.

Objective:To investigate the relationship between serum endothelia cell-specific molecules (endocan), 8-hydroxy-2′deoxyguanosine (8-OHdG) and restless legs syndrome (RLS) in patients with end-stage renal disease (ESRD) on hemodialysis.Methods:A total of 180 ESRD hemodialysis patients admitted to the Shenyang First People′s Hospital, the Liaoning Electric Power Center Hospital and the Northern Theater Command General Hospital of the Chinese People′s Liberation Army from May 2021 to May 2023 were prospectively selected and divided into RLS group (41 cases) and non-RLS group (139 cases) according to whether RLS occurred. According to the International RLS Rating Scale (IRLS), the patients with RLS were divided into mild group (12 cases), moderate group (19 cases) and severe group (10 cases). Serum endocan and 8-OHdG levels were detected by enzyme-linked immunosorbent assay. Pearson correlation was used to analyze the correlation between serum endocan, 8-OHdG levels and IRLS scores in RLS patients. Multivariate logistic regression model was used to evaluate the influencing factors of RLS in ESRD hemodialysis patients. The diagnostic value of serum endocan and 8-OHdG levels in ESRD hemodialysis patients was analyzed by receiver operating characteristic (ROC) curve.Results:Comparing with the non-RLS group [(4.03±1.23)ng/ml, (0.38±0.13)ng/ml], the levels of serum endocan in the RLS group [(6.10±1.95)ng/ml], 8-OHdG [(0.53±0.14)ng/ml] were higher ( t=8.180, 6.379, P<0.001). The levels of serum endocan of the RLS patients in the mild, moderate and severe groups [(3.61±1.01)ng/ml, (6.39±1.41)ng/ml, (8.54±2.68)ng/ml], 8-OHdG [(0.39±0.07)ng/ml, (0.51±0.05)ng/ml, (0.74±0.08)ng/ml] were increased successively ( F=23.065, 83.013, P<0.001). Pearson correlation analysis showed that serum endocan and 8-OHdG levels were positively correlated with IRLS scores in RLS patients ( r=0.813, 0.830, P<0.001). Multivariate logistic regression analysis showed that long dialysis time, elevated serum endocan and 8-OHdG were independent risk factors for RLS in ESRD hemodialysis patients, and elevated ferritin was independent protective factor (all P<0.05). ROC curve analysis showed that the area under the curve of RLS in ESRD hemodialysis patients diagnosed with serum endocan and 8-OHdG combined was 0.919 (95% CI: 0.870-0.955), which was larger than in patients diagnosed with serum endocan and 8-OHdG alone 0.791 (95% CI: 0.724-0.848), 0.780 (95% CI: 0.712-0.838). Conclusions:The increase of serum endocan and 8-OHdG is related to the occurrence and severity of RLS in ESRD hemodialysis patients. The combined detection of serum endocan and 8-OHdG levels is of high diagnostic value in the occurrence of RLS in ESRD hemodialysis patients.

Objective:To explore the impacts of remimazolam(REM)on the proliferation,migration,and invasion of hepatocellular carcinoma cells by regulating sphingosine kinase 1/sphingosine 1-phosphate/sphingosine 1-phosphate receptor 3(SPHK1/S1P/S1PR3)signaling pathway.Methods:Human hepatocellular carcinoma cell line HepG2 was treated with REM at concentrations of 0,20,40,80,160,and 320 μmol/L respectively.Cell survival rate was measured,and drug concentrations were screened.Based on the results of cell survival rate,the logarithmic phase cells were di-vided into five groups:Control group,low,medium,and high concentrations of remifentanil groups(REM-L,REM-M,REM-H groups;20,40 and 80 μmol/L),and high concentrations of remifentanil+SPHK1 activator group(REM-H+K6PC-5 group).MTT assay was used to detect the survival rate of HepG2 cells.Plate cloning experiment was performed to de-tect cell proliferation ability.Scratch experiment was performed to detect cell migration ability.Transwell chamber method was performed to detect cell invasion ability.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect the SPHK1,S1P,S1PR3,Bax,and Bcl-2 proteins in cells.Results:For the Control group,the REM-L,REM-M,and REM-H groups showed that the number of HepG2 cell clones,scratch healing rate,invasion rate,Bcl-2,SPHK1,S1P,S1PR3,and N-cadherin proteins gradually decreased with increasing REM concentration,while the apoptosis rate and Bax,E-cadherin proteins showed an opposite trend(P<0.05).For the REM-H group,the REM-H+K6PC-5 group showed a clear increase in the number of HepG2 cell clones,scratch healing rate,invasion rate,Bcl-2,SPHK1,S1P,S1PR3,and N-cadherin proteins,and a clear decrease in the apoptosis rate and Bax,E-cadherin proteins(P<0.05).Conclusion:REM may inhibit the proliferation,migration,and invasion of hepatocellular carcinoma cells and promote cell apoptosis by suppressing SPHK1/S1P/S1PR3 signaling pathway.

Objective:To investigate the relationship between serum endothelia cell-specific molecules (endocan), 8-hydroxy-2′deoxyguanosine (8-OHdG) and restless legs syndrome (RLS) in patients with end-stage renal disease (ESRD) on hemodialysis.Methods:A total of 180 ESRD hemodialysis patients admitted to the Shenyang First People′s Hospital, the Liaoning Electric Power Center Hospital and the Northern Theater Command General Hospital of the Chinese People′s Liberation Army from May 2021 to May 2023 were prospectively selected and divided into RLS group (41 cases) and non-RLS group (139 cases) according to whether RLS occurred. According to the International RLS Rating Scale (IRLS), the patients with RLS were divided into mild group (12 cases), moderate group (19 cases) and severe group (10 cases). Serum endocan and 8-OHdG levels were detected by enzyme-linked immunosorbent assay. Pearson correlation was used to analyze the correlation between serum endocan, 8-OHdG levels and IRLS scores in RLS patients. Multivariate logistic regression model was used to evaluate the influencing factors of RLS in ESRD hemodialysis patients. The diagnostic value of serum endocan and 8-OHdG levels in ESRD hemodialysis patients was analyzed by receiver operating characteristic (ROC) curve.Results:Comparing with the non-RLS group [(4.03±1.23)ng/ml, (0.38±0.13)ng/ml], the levels of serum endocan in the RLS group [(6.10±1.95)ng/ml], 8-OHdG [(0.53±0.14)ng/ml] were higher ( t=8.180, 6.379, P<0.001). The levels of serum endocan of the RLS patients in the mild, moderate and severe groups [(3.61±1.01)ng/ml, (6.39±1.41)ng/ml, (8.54±2.68)ng/ml], 8-OHdG [(0.39±0.07)ng/ml, (0.51±0.05)ng/ml, (0.74±0.08)ng/ml] were increased successively ( F=23.065, 83.013, P<0.001). Pearson correlation analysis showed that serum endocan and 8-OHdG levels were positively correlated with IRLS scores in RLS patients ( r=0.813, 0.830, P<0.001). Multivariate logistic regression analysis showed that long dialysis time, elevated serum endocan and 8-OHdG were independent risk factors for RLS in ESRD hemodialysis patients, and elevated ferritin was independent protective factor (all P<0.05). ROC curve analysis showed that the area under the curve of RLS in ESRD hemodialysis patients diagnosed with serum endocan and 8-OHdG combined was 0.919 (95% CI: 0.870-0.955), which was larger than in patients diagnosed with serum endocan and 8-OHdG alone 0.791 (95% CI: 0.724-0.848), 0.780 (95% CI: 0.712-0.838). Conclusions:The increase of serum endocan and 8-OHdG is related to the occurrence and severity of RLS in ESRD hemodialysis patients. The combined detection of serum endocan and 8-OHdG levels is of high diagnostic value in the occurrence of RLS in ESRD hemodialysis patients.