1.Construction,prokaryotic expression and in vitro TLR5 activity assay of Esche-richia coli Nissle 1917 flagellin's hypervariable region deletion
Guixian ZHOU ; Shihui WU ; Minle WANG ; Yixiao LIAO ; Shuang LI ; Zemin YANG ; Ying YANG
Chinese Journal of Veterinary Science 2025;45(3):449-457
The structure and potential antigenic epitopes of FliCEcN were analysed using bioinformat-ics technology.With the help of ClonExpress? homologous recombination technology,primers were designed to deletion of different structural domains in the highly variable region of FliCEcN and cloned into pET-28a(+)expression vector for expression.The expressed flagellin variants were identified by SDS-PAGE and Western blot.Endotoxin residues in the flagellin variants were detected by horseshoe crab reagent chromatography,and Caco-2 cells were stimulated with differ-ent concentrations of flagellin variants.The biological activity of each flagellin variant was assessed by detecting the secretion level of IL-8.Bioinformatic analysis showed that most of the structural domains in the highly variable region of FliCEcN were predicted to contain potential antigenic epitopes.PC R results showed that fliC△820-1 518,fliC△736-963,fliC△985-1 200,fliC △748-828,fliC△1 114-1 191,andfliC△1 225-1311 were approximately 1 095,1 566,1 578,1 713,1 716 and 1 707 bp,respectively.SDS-PAGE results showed that the sizes of the flagellin variants treated with nickel column purifi-cation and dialysis replication were about 41.36,57.06,57.50,61.97,61.95 and 61.56 kDa,respec-tively.Western blot results showed that all six flagellin variants reacted specifically with anti-His monoclonal antibody and E.coli H7 antigen diagnostic serum.The results of TLR5 activity assay showed that the flagellin variants missing different structural domains retained their TLR5 agonist function.In this study,six flagellin variants with different structural domains of FliCEcN deletion hypervariable region were successfully constructed,and all of them retained their TLR5 agonist function and showed good biological properties.This provides a reference for further research on the adjuvant effect of flagellin after deletion of different structural domains and the effect of flagel-lin antibody titer on its adjuvant effect.
2.Isolation,identification and genome-wide analysis of a strain of Klebsiella pneu-moniae type ST-4263 from Kole pigs
Guixian ZHOU ; Shihui WU ; Minle WANG ; Yixiao LIAO ; Shuang LI ; Zemin YANG ; Ming WEN ; Simei XIAN ; Ying YANG
Chinese Journal of Veterinary Science 2025;45(8):1679-1687,1695
The 16S rRNA sequencing,whole genome sequencing and drug sensitivity tests were used to identify the isolates molecularly and to detect and analyse their virulence genes,resistance genes and drug resistance.The results showed that the isolate was highly homologous to Klebsiella pneumoniae X4 and located on the same branch by 16S rRNA sequence analysis,and it was named as KLKp10.Whole genome sequencing results showed that the KLKp10 genome was 5 342 841 bp in length,containing 5 138 genes,346 repetitive segments,6 rRNAs and 81 tRNAs,with a GC con-tent of 57.30%.MLST analysis showed that KLKp10 belongs to the ST-4263 type.The functions of 4 097 of the genes encoding proteins were classified and annotated by COG,and there were also 382 genes with unknown functions.A total of 50 functional classifications were involved in the an-notation results based on the GO database;33 kinds of signaling pathways were covered based on the signaling pathway annotations in the KEGG database.A total of 443 virulence genes were screened in the VFDB database,of which 339 belonged to the Set A database and could encode 124 virulence factors.The 101 resistance genes were predicted by comparing with the CARD database,among which there were more resistance genes against β-lactam antibiotics.The results of drug sensitivity test showed that KLKp10 was highly sensitive to ceftazidime,gentamicin,azithro-mycin,chloramphenicol,norfloxacin,ofloxacin,and enrofloxacin;moderately sensitive to ceftriax-one,neomycin,kanamycin,and streptomycin;and resistant to ciprofloxacin,tetracycline,amoxicil-lin,and penicillin.In this study,we systematically revealed the gene-wide characterization,virulence factors and drug resistance of Klebsiella pneumoniae KLKp10 of Kole pig origin,which provides important data support for the study of Klebsiella pneumoniae at the overall level of its genome.
3.Validity and reliability of the Mentalization Questionnaire(MZQ)in Chinese college students
Zemin ZHOU ; Qian HUANG ; Min CHEN ; Gang YUAN ; Xiang LI ; Xuemei LI
Chinese Mental Health Journal 2025;39(3):287-292
Objective:To examine the validity and reliability of the Chinese version of the Mentalization Questionnaire(MZQ)in a sample of Chinese college students.Methods:Totally 3 985 college students were select-ed and randomly divided into Sample 1(n=1 992)and Sample 2(n=1 993).Sample 1 was used to test explorato-ry factor analysis,Sample 2 was used to test criterion-related validity and internal consistency reliability.Totally 596 students were randomly selected from Sample 2,which was subjected to validation factor analysis.After 1 week of initial testing,85 individuals were randomly selected from the total sample for retesting.The Reflective Functioning Questionnaire(RFQ-8)was used to test criterion-related validity.Results:Multiple exploratory factor analyses were conducted on the single-factor structure,and 9 items were finally retained.Validation factor analyses indicated a good fit of the single-factor structure(x2=131.01,x2/df=4.83,P<0.001,NFI=0.94,CFI=0.95,GFI=0.95,IFI=0.95,TLI=0.93,RMSEA=0.08).The Chinese version of the MZQ scores were negatively correlated with the RFQ-C scores(r=-0.59,P<0.01),and positively correlated with the RFQ-U scores(r=0.28,P<0.01).The Cronbach α coefficient of the Chinese version of MZQ was 0.89,and the retest reliability(ICC)was 0.82.Conclusion:The Chinese version of the Mentalization Questionnaire(MZQ)has ideal validity and reliability in Chinese college students.
4.Isolation,identification and genome-wide analysis of a strain of Klebsiella pneu-moniae type ST-4263 from Kole pigs
Guixian ZHOU ; Shihui WU ; Minle WANG ; Yixiao LIAO ; Shuang LI ; Zemin YANG ; Ming WEN ; Simei XIAN ; Ying YANG
Chinese Journal of Veterinary Science 2025;45(8):1679-1687,1695
The 16S rRNA sequencing,whole genome sequencing and drug sensitivity tests were used to identify the isolates molecularly and to detect and analyse their virulence genes,resistance genes and drug resistance.The results showed that the isolate was highly homologous to Klebsiella pneumoniae X4 and located on the same branch by 16S rRNA sequence analysis,and it was named as KLKp10.Whole genome sequencing results showed that the KLKp10 genome was 5 342 841 bp in length,containing 5 138 genes,346 repetitive segments,6 rRNAs and 81 tRNAs,with a GC con-tent of 57.30%.MLST analysis showed that KLKp10 belongs to the ST-4263 type.The functions of 4 097 of the genes encoding proteins were classified and annotated by COG,and there were also 382 genes with unknown functions.A total of 50 functional classifications were involved in the an-notation results based on the GO database;33 kinds of signaling pathways were covered based on the signaling pathway annotations in the KEGG database.A total of 443 virulence genes were screened in the VFDB database,of which 339 belonged to the Set A database and could encode 124 virulence factors.The 101 resistance genes were predicted by comparing with the CARD database,among which there were more resistance genes against β-lactam antibiotics.The results of drug sensitivity test showed that KLKp10 was highly sensitive to ceftazidime,gentamicin,azithro-mycin,chloramphenicol,norfloxacin,ofloxacin,and enrofloxacin;moderately sensitive to ceftriax-one,neomycin,kanamycin,and streptomycin;and resistant to ciprofloxacin,tetracycline,amoxicil-lin,and penicillin.In this study,we systematically revealed the gene-wide characterization,virulence factors and drug resistance of Klebsiella pneumoniae KLKp10 of Kole pig origin,which provides important data support for the study of Klebsiella pneumoniae at the overall level of its genome.
5.Construction,prokaryotic expression and in vitro TLR5 activity assay of Esche-richia coli Nissle 1917 flagellin's hypervariable region deletion
Guixian ZHOU ; Shihui WU ; Minle WANG ; Yixiao LIAO ; Shuang LI ; Zemin YANG ; Ying YANG
Chinese Journal of Veterinary Science 2025;45(3):449-457
The structure and potential antigenic epitopes of FliCEcN were analysed using bioinformat-ics technology.With the help of ClonExpress? homologous recombination technology,primers were designed to deletion of different structural domains in the highly variable region of FliCEcN and cloned into pET-28a(+)expression vector for expression.The expressed flagellin variants were identified by SDS-PAGE and Western blot.Endotoxin residues in the flagellin variants were detected by horseshoe crab reagent chromatography,and Caco-2 cells were stimulated with differ-ent concentrations of flagellin variants.The biological activity of each flagellin variant was assessed by detecting the secretion level of IL-8.Bioinformatic analysis showed that most of the structural domains in the highly variable region of FliCEcN were predicted to contain potential antigenic epitopes.PC R results showed that fliC△820-1 518,fliC△736-963,fliC△985-1 200,fliC △748-828,fliC△1 114-1 191,andfliC△1 225-1311 were approximately 1 095,1 566,1 578,1 713,1 716 and 1 707 bp,respectively.SDS-PAGE results showed that the sizes of the flagellin variants treated with nickel column purifi-cation and dialysis replication were about 41.36,57.06,57.50,61.97,61.95 and 61.56 kDa,respec-tively.Western blot results showed that all six flagellin variants reacted specifically with anti-His monoclonal antibody and E.coli H7 antigen diagnostic serum.The results of TLR5 activity assay showed that the flagellin variants missing different structural domains retained their TLR5 agonist function.In this study,six flagellin variants with different structural domains of FliCEcN deletion hypervariable region were successfully constructed,and all of them retained their TLR5 agonist function and showed good biological properties.This provides a reference for further research on the adjuvant effect of flagellin after deletion of different structural domains and the effect of flagel-lin antibody titer on its adjuvant effect.
6.Validity and reliability of the Mentalization Questionnaire(MZQ)in Chinese college students
Zemin ZHOU ; Qian HUANG ; Min CHEN ; Gang YUAN ; Xiang LI ; Xuemei LI
Chinese Mental Health Journal 2025;39(3):287-292
Objective:To examine the validity and reliability of the Chinese version of the Mentalization Questionnaire(MZQ)in a sample of Chinese college students.Methods:Totally 3 985 college students were select-ed and randomly divided into Sample 1(n=1 992)and Sample 2(n=1 993).Sample 1 was used to test explorato-ry factor analysis,Sample 2 was used to test criterion-related validity and internal consistency reliability.Totally 596 students were randomly selected from Sample 2,which was subjected to validation factor analysis.After 1 week of initial testing,85 individuals were randomly selected from the total sample for retesting.The Reflective Functioning Questionnaire(RFQ-8)was used to test criterion-related validity.Results:Multiple exploratory factor analyses were conducted on the single-factor structure,and 9 items were finally retained.Validation factor analyses indicated a good fit of the single-factor structure(x2=131.01,x2/df=4.83,P<0.001,NFI=0.94,CFI=0.95,GFI=0.95,IFI=0.95,TLI=0.93,RMSEA=0.08).The Chinese version of the MZQ scores were negatively correlated with the RFQ-C scores(r=-0.59,P<0.01),and positively correlated with the RFQ-U scores(r=0.28,P<0.01).The Cronbach α coefficient of the Chinese version of MZQ was 0.89,and the retest reliability(ICC)was 0.82.Conclusion:The Chinese version of the Mentalization Questionnaire(MZQ)has ideal validity and reliability in Chinese college students.
7.A Validity Analysis of the Measurement of Depression and Anxiety in Employees
Feixiang ZHOU ; Jing LIU ; Zemin LIU
Chinese Journal of Health Statistics 2024;41(3):349-353
Objective To evaluate the validity of the Patient Health Questionnaire(PHQ-9)and the Generalized Anxiety Disorder Scale(GAD-7)in measuring depression and anxiety among employees.Methods A stratified random sample of 1117 employees from various departments of an enterprise was included in the study.The item analysis was performed using the Grade Response Model in IRT to assess the dimensionality,discrimination,difficulty coefficient and average information of each item.The model fit and differential item functioning were also assessed.Results The study population consisted of 661(59.2%)employees under the age of 45 and 456(40.8%)employees aged 45 and above;802(71.8%)were male and 315(28.2%)were female.The Cronbach's α coefficients for the PHQ-9 and GAD-7 were 0.923 and 0.951,and the split-half reliability coefficients were 0.863 and 0.940.The ratio of the first characteristic root to the second characteristic root of both the PHQ-9 and GAD-7 scales was>3,in line with the hypothesis of unidimensionality.The discrimination of the PHQ-9 items ranged from 2.362 to 3.618,the difficulty coefficient from-0.949 to 2.639,and the average information from 0.772 to 1.935.The discrimination of the GAD-7 items ranged from 3.631 to 5.580,the difficulty coefficient from-0.469 to 2.044,and the average information from 1.880 to 3.547.Differential item functioning indicated that age and gender did not influence the screening of the two scales.Conclusion The PHQ-9 and GAD-7 scales can be used as psychometric tools to assess depression and generalized anxiety disorder in corporate employees.
8.Clinical application of airway diagnosis procedures in patients with multifactor suspicious difficult airway with complex mouth restriction
Wanhua ZHOU ; Xiaofei WANG ; Zemin XUN
Journal of Chinese Physician 2024;26(10):1530-1534
Objectives:To analyze the safety and clinical effect of airway diagnosis in multifactor suspicious difficult airway patients with complex mouth restriction after entering the operating room.Methods:From March 2018 to May 2023, a total of 21 patients with suspicious difficult airway with complex mouth restriction were collected from the Northwest Women′s and Children′s Hospital and the Third Affiliated Hospital of Air Force Military Medical University, who were evaluated according to LEMON airway evaluation rule. Ventilation capacity score, airway airway score with soft vision mirror and comprehensive airway score of difficult airway were performed after admission to the operating room. Patients diagnosed with non-difficult airway were divided into group N and patients diagnosed with difficult airway were divided into group D. The number and proportion of patients in the two groups were counted, and the process of airway diagnosis, the way of airway establishment, the results, the frequency of hypoventilation during airway establishment, the number of intubation, and the time of airway establishment were analyzed.Results:A total of 15 patients in group N, accounting for 71.4%, underwent rapid intravenous induction nasal intubation after diagnosis of non-difficult airway, and the intubation was successful all the time. No hypoventilation occurred in all patients during intubation. A total of 6 patients in the group D, accounting for 28.6%, were assisted by pure oxygen mask ventilation after diagnosis of difficult airway, and underwent awake nasotracheal intubation after recovery. 4 patients were successfully intubated once, 2 patients were successfully intubated twice, and 1 patient experienced one hypopnea during the evaluation of ventilation capacity. The airway establishment time of the group N was (9.3±0.8)min, and that of the group D was (20.0±2.0)min, the difference was statistically significant ( P<0.05). The airway establishment time of all patients was (12.0±5.0)min, and the awake intubation time was (12.0±0.7.0)min. The proportion of patients satisfied with anesthesia in the group N was higher than that in the group D, and the difference was statistically significant ( P<0.05). No agitation during sevoflurane induction or recovery was observed in both groups. Conclusions:The diagnosis procedure of difficult airway can be safely applied to patients with multi-factor suspicious difficult airway with complex mouth restriction. Most of these patients do not have truly difficult airways; The application of difficult airway diagnosis procedures in these patients can significantly reduce unnecessary awake endotracheal intubation; Compared with the direct choice of awake intubation, following the difficult airway diagnosis procedure does not increase airway establishment time in such patients.
9.Identification of lipid droplets in gut bacteria.
Kai ZHANG ; Chang ZHOU ; Zemin LI ; Xuehan LI ; Ziyun ZHOU ; Linjia CHENG ; Ahmed Hammad MIRZA ; Yumeng SHI ; Bingbing CHEN ; Mengwei ZHANG ; Liujuan CUI ; Congyan ZHANG ; Taotao WEI ; Xuelin ZHANG ; Shuyan ZHANG ; Pingsheng LIU
Protein & Cell 2023;14(2):143-148
10.Application of behaviour change wheel theory in management of cardiovascular disease
Ziwei YOU ; Zemin KUANG ; Yanhui ZHOU ; Lirong WU ; Hongjuan HU
Chinese Journal of Modern Nursing 2021;27(1):121-127
Cardiovascular disease is currently the leading cause of death all over the world, and its prevalence and mortality are still rising. Changing risk health behaviors is an important prevention and treatment strategy for cardiovascular diseases, which can effectively delay the course of disease and improve prognosis of patients. However, the existing chronic disease management model does not fully exploit and utilize intervenable factors to maximize the effect of intervention. Based on 19 behavioral intervention theoretical frameworks, the behaviour change wheel (BCW) provides clear guidance for behavioral analysis and intervention design, which is worthy of in-depth study and application. This article reviews the framework content, implementation steps and application status of BCW in cardiovascular disease management at home and abroad, in order to provide theoretical support and practical guidance for domestic application of BCW theory to conduct behavior intervention.

Result Analysis
Print
Save
E-mail