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.

Lymph node metastasis (LNM) is a crucial risk factor influencing an unfavorable prognosis in specific cancers. Fundamental research illuminates our understanding of tumor behavior and identifies valuable therapeutic targets. Nevertheless, the exploration of fundamental theories and the validation of clinical therapies hinge on preclinical experiments. Preclinical models, in this context, serve as the conduit connecting fundamental theories to clinical outcomes. In vivo models established in animals offer a valuable platform for comprehensively observing interactions between tumor cells and organisms. Using various experimental animals, including mice, diverse methods, such as carcinogen-induced tumorigenesis, tumor cell line or human tumor transplantation, genetic engineering, and humanization, have been used effectively to construct numerous models for tumor LNM. Carcinogen-induced models simulate the entire process of tumorigenesis and metastasis. Transplantation models, using human tumor cell lines or patient-derived tumors, offer a research platform closely mirroring the histology and clinical behavior of human tumors. Genetically engineered models have been used to delve into the mechanisms of primary tumorigenesis within an intact microenvironment. Humanized models are used to overcome barriers between human and murine immune systems. Beyond mouse models, various other animal models have unique advantages and limitations, all contributing to exploring LNM. This review summarizes existing in vitro and animal preclinical models, identifies current bottlenecks in preclinical research, and offers an outlook on forthcoming preclinical models.
Animals ; Humans ; Mice ; Lymphatic Metastasis/pathology* ; Disease Models, Animal ; Cell Line, Tumor

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

Objective To investigate the implementation of surveillance,prevention and control measures for healthcare-associated infection(HAI)in maternal and child healthcare(MCH)institutions,and provide policy evi-dence for optimizing HAI prevention and control in MCH institutions.Methods Stratified sampling was conducted among the MCH institutions at provincial,municipal and county levels in 8 provinces/autonomous regions.A uni-fied questionnaire was designed and the online survey was conducted through"Questionnaire Star".Results The data from 123 MCH institutions were included in the analysis.90.24％of the MCH institutions carried out compre-hensive surveillance on HAI.The ratios of MCH institutions which implemented targeted surveillance on HAI in neonatal intensive care unit(NICU),surgical site infection,multidrug-resistant organisms(MDROs)and HAI in intensive care units(non-NICU excluded)were 89.66％,85.96％,80.77％,and 74.19％,respectively.51.22％MCH institutions adopted information surveillance system on HAI cases.94.31％MCH institutions carried out surveillance on hand hygiene compliance.Over 90％MCH institutions carried out surveillance on environment hy-giene in high-risk departments.71.54％MCH institutions conducted centralized cleaning,disinfection,sterilization and supply for reusable medical instruments in the central sterile supply department(CSSD).Over 90％MCH insti-tutions established three-level pre-examination triage systems.86.18％set up transitional wards.MCH institutions generally adopted a management model with established effective communication,full appointment visits,and sepa-rate visits for special medical groups,such as registered pregnant women,high-risk newborns,healthcare groups,and long-term rehabilitation patients.However,the ratio of institutions conducting on-line follow-up visits was less than 50％.Conclusion MCH institutions have generally carried out comprehensive and targeted surveillance on HAI.Information surveillance need to be facilitated.Hand hygiene and environmental hygiene surveillance has been popularized to a certain extent at all levels of MCH institutions.The cleaning,disinfection,sterilization,and supply processes of reusable medical devices in a few MCH institutions are not standardized.Special medical populations get effective management.On-line healthcare is to be further promoted.

Objective:To identify the risk factors for prolonged length of stay in post-anesthesia care unit (PACU-LOS) and development of a prediction model in the patients undergoing radical esophagectomy.Methods:The medical records from patients of both sexes, aged 40-80 yr, of American Society of Anesthesiologists Physical Status classificationⅠ-Ⅲ, transferred to PACU with tracheal intubation after radical esophagectomy under general anesthesia in our hospital from January 2019 to December 2020, were retrospectively collected. The patient′s age, gender, American Society of Anesthesiologists Physical Status classification, smoking history, drinking history, history of non-thoracic surgery, history of hypertension, history of diabetes mellitus, preoperative anemia, respiratory diseases, doses of anesthetics, preoperative nerve block, intraoperative consumption of opioids and dexmedetomidine, operation method (thoracotomy and endoscopic surgery), operation time, usage of vascular drugs, bradycardia, hypotension, red blood cell infusion, plasma infusion, total infusion volume, blood loss and urine volume were collected. The extubation time in PACU, visual analog scale scores at rest at 10 min after extubation, consumption of rescue analgesics in PACU, hypoxemia after extubation, and occurrence of nausea and vomiting were also collected. Patients were divided into PACU-LOS normal group (PACU-LOS≤2 h) and PACU-LOS prolonged group (PACU-LOS>2 h) according to the PACU-LOS. Logistic regression analysis was used to identity the risk factors for prolonged PACU-LOS in the patients undergoing radical esophagectomy, and the predictive model was established and verified. The receiver operating characteristic curves were used to evaluate the model discrimination and Hosmer-Lemshow goodness-of-fit test was used to evaluate the consistency of the model.Results:A total of 943 patients were included in this study, and the incidence of prolonged PACU-LOS was 15.7%. The results of logistic regression analysis showed that chronic obstructive pulmonary disease ( OR=4.900, 95% confidence interval [ CI] 2.512-9.556), increasing age ( OR=22.154, 95% CI 6.736-73.003), prolonged time of extubation ( OR=1.214, 95% CI 1.174-1.256) and hypoxemia after extubation ( OR=4.891, 95% CI 2.167-11.039) were risk factors for prolonged PACU-LOS, and the preoperative use of nerve block ( OR=0.358, 95% CI 0.190-0.672) was a protective factor for prolonged PACU-LOS in the patients undergoing radical esophagectomy ( P<0.05). The area under the receiver operating characteristic curve (95% CI) was 0.947 (0.925-0.963), the sensitivity was 0.878, and the specificity was 0.906. The internal validation of the prediction model was carried out using the receiver operating characteristic curve in the validation set, and the area under the curve (95% CI) was 0.942 (0.895-0.942, P<0.001) and the Youden index was 0.784. The line chart prediction model was developed. The prediction analysis model was verified by Hosmer-Lemshow test, P<0.001, and the C-index visualized line chart prediction model was 0.946. Conclusions:Preoperative chronic obstructive pulmonary disease, increasing age, prolonged time of extubation and hypoxemia after extubation are risk factors for prolonged PACU-LOS, and preoperative use of nerve block is a protective factor for prolonged PACU-LOS. The risk prediction model developed can effectively predict the occurrence of prolonged PACU-LOS in the patients undergoing radical esophagectomy.

ObjectiveTemporal heterogeneity in lung cancer presents as fluctuations in the biological characteristics, genomic mutations, proliferation rates, and chemotherapeutic responses of tumor cells over time, posing a significant barrier to effective treatment. The complexity of this temporal variance, coupled with the spatial diversity of lung cancer, presents formidable challenges for research. This article will pave the way for new avenues in lung cancer research, aiding in a deeper understanding of the temporal heterogeneity of lung cancer, thereby enhancing the cure rate for lung cancer. MethodsRaman spectroscopy emerges as a powerful tool for real-time surveillance of biomolecular composition changes in lung cancer at the cellular scale, thus shedding light on the disease’s temporal heterogeneity. In our investigation, we harnessed Raman spectroscopic microscopy alongside multivariate statistical analysis to scrutinize the biomolecular alterations in human lung epithelial cells across various timeframes after benzo(a)pyrene exposure. ResultsOur findings indicated a temporal reduction in nucleic acids, lipids, proteins, and carotenoids, coinciding with a rise in glucose concentration. These patterns suggest that benzo(a)pyrene induces structural damage to the genetic material, accelerates lipid peroxidation, disrupts protein metabolism, curtails carotenoid production, and alters glucose metabolic pathways. Employing Raman spectroscopy enabled us to monitor the biomolecular dynamics within lung cancer cells in a real-time, non-invasive, and non-destructive manner, facilitating the elucidation of pivotal molecular features. ConclusionThis research enhances the comprehension of lung cancer progression and supports the development of personalized therapeutic approaches, which may improve the clinical outcomes for patients.

Humans ; Osteoblastoma/surgery* ; Neck/surgery* ; Spinal Neoplasms ; Cervical Vertebrae

Objective:To investigate the application value of ultrasound technology in transurethral enucleation and resection of the prostate(TUERP).Methods:This study included 78 BPH patients admitted in our hospital from June 2021 to June 2023,aged 70.68±8.63 years and with the indication of surgery.We randomly divided them into two groups to receive TUERP(the control group,n=39)and ultrasound-assisted TUERP(the US-TUERP group,n=39).We statistically analyzed and compared the rele-vant parameters obtained before and after operation between the two groups.Results:No statistically significant differences were ob-served in the operation time and bladder irrigation time between the two groups(P>0.05).More glandular tissues were removed but less intraoperative bleeding and fewer perioperative complications occurred in the US-TUERP group than in the control.Compared with the baseline,IPSS,postvoid residual urine volume(PVR),quality of life score(QOL)and maximum urinary flow rate(Qmax)were significantly improved in both groups at 1 and 3 months after surgery,even more significantly in the US-TUERP than in the control group(P<0.05).Conclusion:US-TUERP helps achieve complete resection of the hyperplastic prostatic tissue along the surgical capsule at the anatomical level,with a higher safety,fewer perioperative complications,and better therapeutic effects.

Objective:To investigate the clinical application value of injection of indocyanine green(ICG)via vasopuncture in fluorescence laparoscopic radical prostatectomy(FLRP).Methods:We retrospectively analyzed the clinical data on 50 cases of PCa treated by injection of ICG via vasopuncture in FLRP.The patients were aged(70.60±5.67)years old,with an average PSA value of(18.42±2.69)μg/L.During the operation,we injected ICG at 0.5 ml by vasopuncture through the vas deferens at each side of the scrotum,observed the visualized images of the vas deferens and seminal vesicles using normal high-definition,black-and-white fluorescence,green fluorescence,and color fluorescence respectively,and then isolated the adherent seminal vesicles under the laparoscope.Results:A total of 93 injections of ICG were completed,86 bilaterally,4 on the right and 3 on the left.The vas defer-ens and seminal vesicles were visualized in 41 cases(60 sides,64.52%),19 bilaterally,7 on the right and 15 on the left.Spillage of the fluorescent agent occurred in 9 cases during the incision of the bladder neck and adhesion of the seminal vesicles was found intra-operatively in 10 cases,in which the seminal vesicles were all quickly located by fluorescence visualization.No rectal injury occurred during the surgery.Mild scrotal subcutaneous bruises were observed in 2 cases,with a postoperative pathological Gleason's score of 7.44±0.88.Conclusion:Injection of ICG by vasopuncture is minimally invasive and safe.ICG-mediated near-infrared imaging and real-time fluorescence imaging of the vas deferens and seminal vesicles can achieve precise positioning and removal of the seminal vesicles and prostate gland without causing rectal injury.

Objective:To evaluate the safety and efficiency of China-made Toumai Robot-assisted laparoscopic radical prosta-tectomy(LRP).Methods:This study included 40 cases of PCa treated from January 2023 to May 2023 by robot-assisted LRP with preservation of the bladder neck and maximal functional urethral length,15 cases with the assistance of Toumai Robot(the TMR group)and the other 25 with the assistance of da Vinci Robot as controls(the DVR group).We recorded the docking time,laparo-scopic surgery time,vesico-urethral anastomosis time,intraoperative blood loss and postoperative urinary continence,and compared them between the two groups.Results:Operations were successfully completed in all the cases.No statistically significant differ-ences were observed between the TMR and DVR groups in the docking time(6 min vs 5 min,P＞0.05)or intraoperative blood loss(200 ml vs 150 ml,P＞0.05).The TMR group,compared with the DVR group,showed a significantly longer median laparoscopic surgery time(146 min vs 130 min,P＜0.05)and median vesico-urethral anastomosis time(19 min vs 16 min,P＜0.05).There were no statistically significant differences between the TMR and DVR groups in the rates of urinary continence recovery immediately af-ter surgery(60.0％[9/15]vs 64.0％[16/25],P＞0.05)or at 1 month(80.0％[12/15])vs(76.0％[19/25],P＞0.05),3 months(93.3％[14/15])vs(92.0％[23/25],P＞0.05)and 6 months postoperatively(100％[15/15])vs(96％[24/25],P＞0.05).Conclusion:China-made Toumai? Robot surgical system is safe and reliable for laparoscopic radical prosta-tectomy,with satisfactory postoperative recovery of urinary continence.