1.Enzyme-directed Immobilization Strategies for Biosensor Applications
Xing-Bao WANG ; Yao-Hong MA ; Yun-Long XUE ; Xiao-Zhen HUANG ; Yue SHAO ; Yi YU ; Bing-Lian WANG ; Qing-Ai LIU ; Li-He ZHANG ; Wei-Li GONG
Progress in Biochemistry and Biophysics 2025;52(2):374-394
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.
2.Improvement effects of glycyrrhizin on Helicobacter pylori-associated gastritis in rats and its mechanism
Yuhua LIU ; Lian LIU ; Jiuchong WANG ; Dan HUANG ; Sufang ZHOU ; Huanzhi XIAO ; Zhenxiang AN
China Pharmacy 2024;35(2):166-171
OBJECTIVE To investigate the improvement effects of glycyrrhizin (GL) on Helicobacter pylori (HP)-associated gastritis in rats and its mechanism. METHODS HP-associated gastritis rat model was induced by inoculating with 1×109 cfu/mL HP. The model rats were randomly divided into model group, positive control group (HP standard quadruple group), GL low-dose, medium-dose and high-dose groups (5, 20, 50 mg/kg), with 12 rats in each group. Another 12 healthy rats were selected as normal control group. Except the normal control group and model group were given constant volume of normal saline intragastrically, the other groups were given corresponding drugs intragastrically, once a day, for 30 consecutive days. After administration, rats received 13C urea breath test, and delta-over-baseline (DOB) was recorded; the pathological and cellular morphological changes of gastric mucosa in rats were observed, and pathological scoring was performed; the levels of interleukin-8 (IL-8), IL-1β, tumor necrosis factor-α (TNF-α), reactive oxygen species (ROS) and malondialdehyde (MDA) were detected in gastric mucosa of rats; mRNA expressions of high mobility group box-1 protein (HMGB1) and nuclear factor-κ-B (NF-κB), relative expressions of nitric oxide synthases (iNOS) and HMGB1, the phosphorylation level of NF- κBp65 were also detected in rats. RESULTS Compared with normal control group, the DOB value, histopathological score of gastric mucosa, the levels of IL-8, IL-1β, TNF-α, ROS and MDA, relative expressions of HMGB1 and NF- κB mRNA, relative expressions of iNOS and HMGB1 protein and the phosphorylation level of NF-κB p65 were all increased significantly in model group (P<0.05); the epithelial cells of gastric mucosa in rats were incomplete in structure and decreased in the number, with an increase in cell fragments and vacuoles, and significant cell pyknosis. Compared with model group, the changes of the above indexes in GL groups and positive control group were significantly reversed (P<0.05); the changes in the above indicators in the GL high-dose group were more significant than GL low-dose and medium-dose groups (P<0.05); the pathological changes of gastric mucosal cells in rats had all improved. CONCLUSIONS GL may inhibit inflammation and oxidative stress by inhibiting the activation of HMGB1/NF-κB pathway, thus relieving HP-induced gastric mucosal injury.
3.Implementation of surveillance,prevention and control of healthcare-asso-ciated infection in maternal and child healthcare institutions:A nation-wide investigation report
Shuo LI ; Xi YAO ; Hui-Xue JIA ; Wei-Guang LI ; Xun HUANG ; Shu-Mei SUN ; Xi CHENG ; Qing-Lan MENG ; Xiang ZHANG ; Jing-Ping ZHANG ; Ya-Wei XING ; Qing-Qing JIANG ; Lian-Xuan WU ; Bing-Li ZHANG ; Xiao-Jing LIU ; Liu-Yi LI
Chinese Journal of Infection Control 2024;23(3):323-329
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.
4.Clinical results of 100 cases of modified eversion carotid endarterectomy
Leng NI ; Wenzhuo LIAN ; Rong ZENG ; Xiao DI ; Xitao SONG ; Fangda LI ; Liqiang CUI ; Guoqiang SUN ; Yuehong ZHENG ; Changwei LIU
Chinese Journal of Surgery 2024;62(12):1143-1149
Objective:The primary goal of this study is to explore the safety and effectiveness of a new modified eversion carotid endarterectomy (MECEA).Methods:This is a retrospective case series study. One hundred patients were consecutively treated with MECEA by the same operator at Department of Vascular Surgery,Peking Union Medical College Hospital from January 2019 to December 2023. There were 77 males and 23 females. The age was (66.0±8.6)years (range: 39 to 85 years). Twenty-four (24.0%) patients were symptomatic with the degree of carotid stenosis over 50%,76 patients (76.0%) were asymptomatic with the degree of stenosis over 70%. All these patients meet the indication of carotid endarterectomy. The main difference between MECEA and traditional eversion carotid endarterectomy was the anterior,lateral,and posterior walls of the internal carotid artery were incised obliquely from the origin of the internal carotid artery toward the common carotid artery,leaving the wall of internal carotid artery intact at the bifurcation. The surgical process,cardiovascular and cerebrovascular complications and other surgical complications were recorded. The incidences of complications,restenosis of intraoperative target lesions and re-intervention were collected during follow-up.Results:All procedures were performed successfully under general anesthesia. The total operation time was (36.5±10.1)minutes (range: 22 to 65 minutes),and carotid clamping time was (15.0±6.3)minutes (range: 7 to 31 minutes). One patient (1.0%) occurred postoperative cerebrovascular accident,1 patient (1.0%) developed cerebral hyperperfusion syndrome (CHS),and another 1 patient (1.0%) suffered myocardial infarction. All these patients were recovered after medical treatment within a week. The follow-up time( M(IQR)) was 24 (28) months (range: 6 to 62 months). Two patients (2.0%) were reported to have hemodynamically significant restenosis within 2 years,with one patient requiring intervention. No patient suffered from ipsilateral ischemic stroke. Conclusions:MECEA is a safe and effective surgical method of treating carotid artery stenosis. This method can reduce carotid clamping time and lowers the risk of ischemic stroke. Meantime,it preserves the integrity of the adventitia at the bifurcation of carotid artery,reduces the chance of restenosis. Moreover,it might be helpful to prevent postoperative CHS due to reducing damage to the carotid body and carotid sinus nerve.
5.Screening and identification of xanthine oxidase inhibitors from Smilax glabra
Hui-Liang ZOU ; Ke CHEN ; Xin-Cai WANG ; Xiao CHENG ; Ling-Jie MENG ; Bai-Lian LIU ; Min CHEN
Chinese Traditional Patent Medicine 2024;46(5):1558-1564
AIM To rapidly screen xanthine oxidase(XOD)inhibitors from Smilax glabra Roxb.by enzyme-immobilized magnetic microspheres and LC-MS/MS,and to confirm the anti-uric acid constituents from S.glabra Roxb.METHODS The immobilized xanthine oxidase was prepared by covalent coupling with carboxyl magnetic beads as a carrier.The xanthine oxidase inhibitors in S.glabra were screened by the specific adsorption of immobilized enzyme.LC-MS/MS and standard substances were used for analysis and comparison,and the inhibitory activity and inhibition type of the screened and identified components were investigated.RESULTS The successful synthesis of immobilized xanthine oxidase was characterized by scanning electron microscopy and infrared spectroscopy.The enzyme loading was 70.50 μg/mg and the relative activity was 79.44%.Thirteen active compounds were screened from the extract of S.glabra,and eleven compounds were identified.The enzyme activity test showed that the inhibitory activites of engeletin and isoengeletin were the strongest,which was close to the positive control allopurinol.The IC50 value and inhibition type were 32.25 μg/mL,mixed inhibition,35.12 μg/mL,competitive inhibition.CONCLUSION The method is simple,rapid,accurate and suitable for directly screened active ingredients which can inhibit XOD from complex extract of traditional Chinese medicines.
6.Application of electrocardiogram in predicting culprit vessels in acute inferior myocardial infarction
Feng LIU ; Hua-Kang LI ; Yan-Xiu CHEN ; Xiao-Lian CAI
Journal of Regional Anatomy and Operative Surgery 2024;33(5):440-444
Objective To explore the value of electrocardiogram in predicting culprit vessels in acute inferior myocardial infarction.Methods The clinical data of 129 patients with acute inferior myocardial infarction admitted to First Affiliated Hospital of Army Medical University from January 2015 to December 2021 were retrospectively analyzed.Patients were received coronary intervention treatment,and the culprit vessels were identified by coronary angiography.The culprit vessels during surgery were recorded.The preoperative electrocardio-gram of patients were analyzed,a new method for predicting culprit vessels in acute inferior myocardial infarction was established based on the previous researches,and the accuracies of the Fiol method,Tiala method,Huang's algorithm and new method in predicting the culprit vessels in acute inferior myocardial infarction were also counted.The predictive abilities of various methods on culprit vessels were evaluated using receiver operating characteristic(ROC)curve and the area under the curve(AUC)was calculated.Results The culprit vessels of 109 patients were the right coronary artery,16 cases were the left circumflex artery,and 4 cases were the anterior descending artery.A total of 52 patients developed clinical complications,of which the culprit vessels of 47 cases were right coronary artery and 5 cases were circumflex artery.The accuracies of the Fiol method,Tierala method,Huang's algorithm,and new method in predicting culprit vessels were 83.7%,81.4%,82.9%,and 85.3%,respectively.The ROC curve analysis showed that the Fiol method(AUC=0.941),Tierrala method(AUC= 0.945),Huang's algorithm(AUC=0.945),and new method(AUC=0.964)all had strong predictive ability for culprit vessels.Conclusion For patients with acute inferior myocardial infarction,the Fiol method,Tierala method,Huang's algorithm,and new method all have high predictive value for culprit vessels in analyzing the preoperative electrocardiogram,and the new method has a higher predictive value.
7.Hydrogen sulfide and neuroinflammation-mediated neurodegenerative diseases
Yu-Lian SHUI ; Zhi-Qiong REN ; Yi-Jie HE ; Bin-Bin CHEN ; Jia HONG ; Ke-Ting LIU ; Li XIAO
Journal of Regional Anatomy and Operative Surgery 2024;33(6):551-554
Hydrogen sulfide,as a third gas signal molecule and neurotransmitter,can play a neuroprotective role by anti-oxidative stress,anti-inflammatory response,metabolic inhibition and other mechanisms.It is of great significance for the occurrence and development of neurodegenerative diseases including Alzheimer's disease(AD)and Parkinson's disease(PD)mediated by neuroinflammation.This article reviews the research progress of hydrogen sulfide and neuroinflammation and its mediated neurodegenerative diseases,so as to provide new ideas for the treatment of neurodegenerative diseases.
8.Dawn of CAR-T cell therapy in autoimmune diseases
Yuxin LIU ; Minghao DONG ; Yunhui CHU ; Luoqi ZHOU ; Yunfan YOU ; Xiaowei PANG ; Sheng YANG ; Luyang ZHANG ; Lian CHEN ; Lifang ZHU ; Jun XIAO ; Wei WANG ; Chuan QIN ; Daishi TIAN
Chinese Medical Journal 2024;137(10):1140-1150
Chimeric antigen receptor (CAR)-T cell therapy has achieved remarkable success in the treatment of hematological malignancies. Based on the immunomodulatory capability of CAR-T cells, efforts have turned toward exploring their potential in treating autoimmune diseases. Bibliometric analysis of 210 records from 128 academic journals published by 372 institutions in 40 countries/regions indicates a growing number of publications on CAR-T therapy for autoimmune diseases, covering a range of subtypes such as systemic lupus erythematosus, multiple sclerosis, among others. CAR-T therapy holds promise in mitigating several shortcomings, including the indiscriminate suppression of the immune system by traditional immunosuppressants, and non-sustaining therapeutic levels of monoclonal antibodies due to inherent pharmacokinetic constraints. By persisting and proliferating in vivo, CAR-T cells can offer a tailored and precise therapeutics. This paper reviewed preclinical experiments and clinical trials involving CAR-T and CAR-related therapies in various autoimmune diseases, incorporating innovations well-studied in the field of hematological tumors, aiming to explore a safe and effective therapeutic option for relapsed/refractory autoimmune diseases.
9.Clinical results of 100 cases of modified eversion carotid endarterectomy
Leng NI ; Wenzhuo LIAN ; Rong ZENG ; Xiao DI ; Xitao SONG ; Fangda LI ; Liqiang CUI ; Guoqiang SUN ; Yuehong ZHENG ; Changwei LIU
Chinese Journal of Surgery 2024;62(12):1143-1149
Objective:The primary goal of this study is to explore the safety and effectiveness of a new modified eversion carotid endarterectomy (MECEA).Methods:This is a retrospective case series study. One hundred patients were consecutively treated with MECEA by the same operator at Department of Vascular Surgery,Peking Union Medical College Hospital from January 2019 to December 2023. There were 77 males and 23 females. The age was (66.0±8.6)years (range: 39 to 85 years). Twenty-four (24.0%) patients were symptomatic with the degree of carotid stenosis over 50%,76 patients (76.0%) were asymptomatic with the degree of stenosis over 70%. All these patients meet the indication of carotid endarterectomy. The main difference between MECEA and traditional eversion carotid endarterectomy was the anterior,lateral,and posterior walls of the internal carotid artery were incised obliquely from the origin of the internal carotid artery toward the common carotid artery,leaving the wall of internal carotid artery intact at the bifurcation. The surgical process,cardiovascular and cerebrovascular complications and other surgical complications were recorded. The incidences of complications,restenosis of intraoperative target lesions and re-intervention were collected during follow-up.Results:All procedures were performed successfully under general anesthesia. The total operation time was (36.5±10.1)minutes (range: 22 to 65 minutes),and carotid clamping time was (15.0±6.3)minutes (range: 7 to 31 minutes). One patient (1.0%) occurred postoperative cerebrovascular accident,1 patient (1.0%) developed cerebral hyperperfusion syndrome (CHS),and another 1 patient (1.0%) suffered myocardial infarction. All these patients were recovered after medical treatment within a week. The follow-up time( M(IQR)) was 24 (28) months (range: 6 to 62 months). Two patients (2.0%) were reported to have hemodynamically significant restenosis within 2 years,with one patient requiring intervention. No patient suffered from ipsilateral ischemic stroke. Conclusions:MECEA is a safe and effective surgical method of treating carotid artery stenosis. This method can reduce carotid clamping time and lowers the risk of ischemic stroke. Meantime,it preserves the integrity of the adventitia at the bifurcation of carotid artery,reduces the chance of restenosis. Moreover,it might be helpful to prevent postoperative CHS due to reducing damage to the carotid body and carotid sinus nerve.
10.Comparison of the risk of gastroesophageal reflux between the lithotomy position and the supine position under SaCoVLM laryngeal mask ventilation
Kun LIU ; Xiao ZHAO ; Chao GONG ; Shuang WU ; Shi-Tong LI ; Lian-Hua CHEN
Fudan University Journal of Medical Sciences 2024;51(3):385-391
Objective To observe whether SaCoVLM laryngeal mask can cause gastroesophageal reflux when the body position changes(lithotomy position and supine position).Methods A total of 70 patients were selected for elective surgery in Shanghai General Hospital from Dec 1st,2021 to Sep 30th,2022.There were 35 patients with ureteroscopy in urology and 35 patients with ankle surgery in trauma orthopedics selected as lithotomy position group and supine position group,respectively.Under SaCoVLM laryngeal mask anesthesia,a dual probe pH electrode was placed in the esophagus to continuously monitor the pH value in the esophagus.The pH values and their changing trends in the lithotomy position and supine position of the esophagus were compared under general anesthesia with SaCoVLM laryngeal mask ventilation,and the possibility of gastroesophageal reflux occurring in both positions was explored.Results The peak airway pressure at each time point(5 min,10 min,15 min,20 min and 30 min)after body position was determined in the two groups,and it in lithotomy position group was significantly higher than that in supine position group,and the difference was statistically significant(P<0.05).The middle and upper esophageal pH values of the two groups were compared 10 min after laryngeal mask insertion,and it in lithotomy position group(6.045±0.490)was significantly lower than the supine group(6.532±0.366),and the difference was statistically significant(P<0.05).The middle and upper esophageal pH values of the two groups were compared 10 min and 15 min after laryngeal mask insertion.They in lithotomy position group were significantly lower than those in supine position group,and the differences were statistically significant(10 min:6.045±0.490 vs.6.532±0.366,P=0.031;15 min:5.828±0.487 vs.6.474±0.411,P=0.048).There was no significant difference in the pH of the middle and upper esophagus between the two groups at 1,5,20 and 30 minutes of laryngeal mask insertion.There was no significant difference in the pH value of the esophageal opening between the two groups at each time point after laryngeal mask insertion.Conclusion Under SaCoVLM laryngeal mask ventilation,the risk of gastroesophageal reflux during lithotomy surgery may be higher than during supine surgery.

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