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.

Alzheimer′s disease is a serious neurodegenerative disorder. Approximately 80% to 90% of patients are accompanied by behavioral and psychological symptoms of dementia (BPSD), which manifest as a series of behavioral, psychological and mental abnormalities. These abnormalities can accelerate the cognitive deterioration and premature death of patients, and thus are regarded as important clinical symptoms. However, the pathogenesis of BPSD is still unknown, and treatment methods are limited. The pathogenesis of BPSD from the perspective of neuroimaging and pathophysiology, and possible treatment measures were analyzed in this article, in order to provide references for the early diagnosis and treatment of BPSD.

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.

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.

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.

Alzheimer′s disease is a serious neurodegenerative disorder. Approximately 80% to 90% of patients are accompanied by behavioral and psychological symptoms of dementia (BPSD), which manifest as a series of behavioral, psychological and mental abnormalities. These abnormalities can accelerate the cognitive deterioration and premature death of patients, and thus are regarded as important clinical symptoms. However, the pathogenesis of BPSD is still unknown, and treatment methods are limited. The pathogenesis of BPSD from the perspective of neuroimaging and pathophysiology, and possible treatment measures were analyzed in this article, in order to provide references for the early diagnosis and treatment of BPSD.

Objective:To evaluate the value of 18F-FDG PET/CT for preoperative localization of epileptogenic foci in refractory epilepsy patients with negative MRI. Methods:Clinical data (550 lobes) of 55 epilepsy patients (38 males, 17 females, age (20.0±8.1) years) with negative MRI who underwent preoperative 18F-FDG PET/CT-MRI between January 2014 and June 2020 at the First Affiliated Hospital of Jinan University were retrospectively analyzed. The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of 18F-FDG PET/CT, video electroencephalogram (VEEG), PET/CT+ VEEG and PET/CT-VEEG for localizing epileptogenic foci were calculated using stereoelectroencephalography (SEEG) and the outcomes of at least 1 year of postoperative follow-up as reference standards. χ2 test was used to compare the efficiencies of different examination modalities for unilobar, multilobar and all patients. Results:The correct lateralization rate of epileptogenic foci was 92.6%(25/27) using PET/CT. The sensitivity, specificity, accuracy, PPV and NPV of PET/CT for localization of epileptogenic foci were 65.1%(54/83), 77.9%(364/467), 76.0%(418/550), 34.4%(54/157) and 92.6%(364/393), respectively. The sensitivities of PET/CT-VEEG for localization of epileptogenic foci in all patients and patients with multilobar epilepsy were higher than those of VEEG alone (75.9%(63/83) vs 45.8%(38/83), 68.6%(35/51) vs 31.4%(16/51); χ2 values: 15.80, 14.16, both P<0.001). The specificities of PET/CT+ VEEG for localization of epileptogenic foci in all patients and patients with unilobar epilepsy were higher than those of VEEG alone (97.6%(456/467) vs 94.6%(442/467), 97.9%(282/288) vs 94.1%(271/288); χ2 values: 5.66, 5.48; P values: 0.017, 0.019). The sensitivity of PET/CT-VEEG (PET/CT and VEEG concordance) for localization of epileptogenic foci was higher than that of PET/CT+ VEEG (PET/CT and VEEG discordance) (8/9 vs 28.4%(21/74); χ2=10.40, P=0.001), and its specificity and accuracy were higher than those of PET/CT-VEEG (PET/CT and VEEG discordance) (93.4%(57/61) vs 71.7%(291/406), 92.9%(65/70) vs 72.1%(346/480); χ2 values: 13.23, 13.96; both P<0.001). Conclusions:18F-FDG PET/CT can localize and lateralize epileptogenic foci in patients with negative MRI. The combination of 18F-FDG PET/CT and VEEG improves the sensitivity, specificity, and accuracy for epileptogenic foci detection. 18F-FDG PET/CT is more accurate in detecting epileptogenic foci when it is concordant with VEEG.

Objective:To explore the effect of visual processing patterns on emotional face processing in patients with Alzheimer's disease (AD).Methods:From June 2020 to August 2021, twenty-two AD patients (AD group) who met the conditions of this study were selected from the memory impairment clinic of the First Affiliated Hospital of Anhui Medical University, and demographically matched twenty-one elderly healthy people (control group) were selected from the patients' family members and community residents. The two groups of subjects performed emotional face visual scanning and facial recognition experiments after completing the evaluation of the cognitive scale and eye movement data were recorded in the emotional face visual scanning task. Statistical analysis of the obtained results was performed using SPSS 23.0 Windows version software. The data that conformed to the normal distribution were tested by independent samples t-test and variance analysis, and the data that did not conform to the normal distribution were tested by nonparametric test. Results:(1)In the emotional face recognition task, the total accuracy of facial emotion recognition of AD patients(0.52(0.42, 0.59)) was lower than that of the normal control group(0.67(0.64, 0.69)), and the difference was statistically significant( Z=-4.023, P<0.01), which was mainly manifested in recognizing complex facial emotion. (2) In the emotional face visual processing task, the saccade count ((1.96±0.97), (2.50±0.44)), fixation count ((3.93±2.58), (6.37±2.08))and fixation time ((1 205.89±727.32)s, (1 761.38±525.54)s)of AD patients were lower than those of the control group( t=-2.314, -3.402, -2.880, all P<0.05), and the surrounding facial fixation time (384.95 (276.51, 587.78)s, 276.06 (190.03, 384.55)s) was higher than that of the control group( Z=-2.478, P=0.013). Patients with AD had a lower fixation count than that in the control group on the eye area of surprise ((3.76±2.90), (6.25±2.19)), anger ((4.48±2.72), (7.06±2.55)) and disgust ((4.10±2.45), (6.67±2.45)), and the differences were statistically significant ( t=-3.164, -3.207, -3.436, all P<0.05). Patients with AD had a lower fixation time than those of the control group on the eye area of surprise ((1 150.26±753.22)s, (1 779.91±551.66)s), angry ((1 430.85±869.52)s, (1 944.51±612.63)s) and disgust ((1 266.14±765.67)s, (1 898.33±676.02)s), and the differences were statistically significant ( t=-3.115, -2.247, -2.865, all P<0.05). (3) Spearman correlation analysis showed that the accuracy of overall emotional face recognition was positively correlated with the fixation time in the eye area in AD patients ( r=0.429, P<0.05). Conclusion:The impaired visual processing of AD patients causes emotional face recognition disorders. Therefore, AD patients have different visual processing patterns in emotional face processing than age-matched normal controls, mainly manifested as the decreased fixation on the eye area.

Objective:To explore whether sleep quality suffers in patients with mild Alzheimer's disease(AD)and mild cognitive impairment(MCI), and to further investigate the correlation between sleep disorders and cognitive function in these patients.Methods:In this study, 30 mild AD patients, 39 MCI patients and 43 demographically matched healthy controls were enrolled.Sleep quality was assessed by the Pittsburgh sleep quality index(PSQI), and cognitive function was assessed by the mini-mental state examination(MMSE), the Montreal cognitive assessment(MoCA)and a set of neuropsychological scales.The correlation of sleep quality with cognitive function was analyzed for the three groups.Results:Differences were significant in sleep time score[0.0(1.0), 1.0(2.0) vs.1.0(1.0), F=8.18, P=0.02]and daytime function score[1.0(1.0), 1.0(1.0) vs.0.0(1.0), F=8.73, P=0.01]between mild AD, MCI and health control groups.Spearman correlation analysis suggested that scores of sleep disorders were negatively correlated with DSB( r=-0.43, P=0.02)and scores of daytime function were positively correlated with ADL( r=0.39, P=0.03)in patients with mild AD.In addition, scores of sleep quality were negatively correlated with the DSB score( r=-0.40, P=0.01), scores of sleep disorders were positively correlated with ADL( r=0.45, P<0.01), scores of daytime function were negatively correlated with DSF( r=-0.42, P=0.01), DSB( r=-0.62, P<0.01)and VFT-S( r=-0.33, P=0.04), and the total PSQI score was negatively correlated with DSF( r=-0.45, P=0.01)and DSB( r=-0.44, P=0.01)in the MCI group. Conclusions:Patients with mild AD and MCI have longer sleep time and impaired daytime function than healthy people, and sleep quality is correlated with memory, attention and daily living ability in patients with mild AD and MCI.

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.