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.

BACKGROUND:Hemivertebra deformity should be treated surgically at an early age,but the risk factors for progression of deformity after hemivertebral resection have not been established.OBJECTIVE:To investigate the curative effect of one-stage posterior hemivertebrae resection and pedicle screw fixation in the treatment of congenital scoliosis,and to further explore the risk factors causing the progression of postoperative deformity.METHODS:The medical records of patients who underwent pedicle screw-rod fixation for unilateral hemivertebral deformity from January 2012 to February 2020 and were followed up for at least 2 years were retrospectively analyzed,and a total of 116 patients met the inclusion criteria.All patients were treated with standing anterior and lateral spinal radiographs taken before,after and at each follow-up time point.The segment Cobb angle,the total scoliosis Cobb angle,the proximal complementary Cobb angle,the distal complementary Cobb angle,and the coronal balance distance,apical vertebra distance,upper instrumented vertebra tilt,upper instrumented vertebra disc angle,lower instrumented vertebra tilt,lower instrumented vertebra disc angle,segmental kyphosis/lordosis,thoracic kyphosis,lumbar lordosis and sagittal vertical axis were measured.The progression of deformity and complications were also recorded.RESULTS AND CONCLUSION:(1)Segment Cobb,total scoliosis Cobb,segmental kyphosis,proximal complementary Cobb,and distal complementary Cobb were significantly corrected after operation(P＜0.05),and remained corrected at the last follow-up.Thoracic kyphosis,lumbar lordosis,coronal balance distance,and sagittal vertical axis were all in the normal range pre-operation,after operation and at the last follow-up.(2)During follow-up,10 patients developed coronary decompensation,which was characterized by abnormal progression.(3)Independent sample t-test showed that preoperative total scoliosis Cobb,preoperative apical vertebra distance,age,Risser sign,postoperative upper instrument vertebra tilt and postoperative lowest instrumented vertebra tilt were correlated with postoperative malformation progression(P＜0.05).(4)Multivariate Logistic regression analysis showed that postoperative lowest instrumented vertebra tilt was an independent risk factor for postoperative malformation progression(P=0.002,OR=1.526).(5)Receiver operating characteristic curve analysis showed that a postoperative lowest instrumented vertebra tilt of 8.14° was the optimal threshold for deformity progression after hemivertebrae resection and pedicle rod fixation(sensitivity 0.900,specificity 0.906,area under curve:0.926).(6)It is indicated that the treatment of congenital scoliosis with one-stage posterior hemivertebrae resection and pedicle nail fixation can achieve satisfactory orthopedic effect.Postoperative lowest instrumented vertebra tilt greater than 8.14° was an independent risk factor for postoperative coronal decompensation.

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.

BACKGROUND:Hemivertebra deformity should be treated surgically at an early age,but the risk factors for progression of deformity after hemivertebral resection have not been established.OBJECTIVE:To investigate the curative effect of one-stage posterior hemivertebrae resection and pedicle screw fixation in the treatment of congenital scoliosis,and to further explore the risk factors causing the progression of postoperative deformity.METHODS:The medical records of patients who underwent pedicle screw-rod fixation for unilateral hemivertebral deformity from January 2012 to February 2020 and were followed up for at least 2 years were retrospectively analyzed,and a total of 116 patients met the inclusion criteria.All patients were treated with standing anterior and lateral spinal radiographs taken before,after and at each follow-up time point.The segment Cobb angle,the total scoliosis Cobb angle,the proximal complementary Cobb angle,the distal complementary Cobb angle,and the coronal balance distance,apical vertebra distance,upper instrumented vertebra tilt,upper instrumented vertebra disc angle,lower instrumented vertebra tilt,lower instrumented vertebra disc angle,segmental kyphosis/lordosis,thoracic kyphosis,lumbar lordosis and sagittal vertical axis were measured.The progression of deformity and complications were also recorded.RESULTS AND CONCLUSION:(1)Segment Cobb,total scoliosis Cobb,segmental kyphosis,proximal complementary Cobb,and distal complementary Cobb were significantly corrected after operation(P＜0.05),and remained corrected at the last follow-up.Thoracic kyphosis,lumbar lordosis,coronal balance distance,and sagittal vertical axis were all in the normal range pre-operation,after operation and at the last follow-up.(2)During follow-up,10 patients developed coronary decompensation,which was characterized by abnormal progression.(3)Independent sample t-test showed that preoperative total scoliosis Cobb,preoperative apical vertebra distance,age,Risser sign,postoperative upper instrument vertebra tilt and postoperative lowest instrumented vertebra tilt were correlated with postoperative malformation progression(P＜0.05).(4)Multivariate Logistic regression analysis showed that postoperative lowest instrumented vertebra tilt was an independent risk factor for postoperative malformation progression(P=0.002,OR=1.526).(5)Receiver operating characteristic curve analysis showed that a postoperative lowest instrumented vertebra tilt of 8.14° was the optimal threshold for deformity progression after hemivertebrae resection and pedicle rod fixation(sensitivity 0.900,specificity 0.906,area under curve:0.926).(6)It is indicated that the treatment of congenital scoliosis with one-stage posterior hemivertebrae resection and pedicle nail fixation can achieve satisfactory orthopedic effect.Postoperative lowest instrumented vertebra tilt greater than 8.14° was an independent risk factor for postoperative coronal decompensation.

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.

Objective:To investigate the spectrum, drug resistance and risk factors of multidrug resistant organism (MDRO) in chronic osteomyelitis.Methods:A retrospective study was conducted by cluster sampling to analyze the 414 patients with chronic osteomyelitis who had been admitted to Department of Orthopaedics, The Second Hospital Affiliated to Shanxi Medical University, Department of Orthopaedics, The People's Hospital of Shanxi Province, Department of Traumatology, Yuncheng City Hospital, and Department of Orthopaedics, Linfen City Hospital from January 2016 to December 2021. The patients were assigned into 2 groups according to whether MDRO had been detected or not. In the MDRO infection group of 150 cases, there were 118 males and 32 females with an age of (48.8±16.2) years; in the non-MDRO infection group of 264 cases, there were 194 males and 70 females with an age of (46.0±17.8) years. The characteristics of bacterial spectrum and drug resistance in MDRO infection were described and analyzed. The 2 groups were compared in terms of clinical data like gender, age, course of disease, body mass index, history of antibiotic use before admission, combined internal diseases, combined trauma, and length of hospital stay. The items with P<0.05 were included in a multivariate logistic regression model to explore the risk factors for MDRO infection. Results:Pathogenic bacteria (331 strains) were detected in 286 of the 414 patients with chronic osteomyelitis, and infection with 168 strains of MDRO was detected in 150 of the 286 patients, yielding a detection rate of 50.8% (168/331). Of the 168 strains of MDR, 129 (76.8%, 129/168) were Gram-positive and 39 (23.2%, 39/168) Gram-negative. Staphylococcus aureus was the most frequently detected species of Gram-positive bacteria (58.1%, 75/129) and the most frequently detected pathogen (44.6%, 75/168) in this study. The resistance of Gram-positive bacteria to peptides and oxazolidinones was low (less than 10%). Multivariate logistic regression analysis showed that age ≥45 years ( OR=6.991, 95% CI: 3.525 to 13.865, P<0.001), essential hypertension ( OR=4.191, 95% CI: 2.070 to 8.485, P<0.001), trauma ( OR=4.232, 95% CI: 2.409 to 7.435, P<0.001) and length of hospital stay ( OR=1.015, 95% CI: 1.001 to 1.029, P=0.030) were the risk factors for MDRO infection in patients with chronic osteomyelitis. Conclusions:The detection rate of MDRO is at a medium to high level. Gram positive bacteria are the main pathogens and resistant to most antibiotics. Antibiotic therapy guided by bacterial culture is of great significance for patients with chronic osteomyelitis. Age≥45 years, essential hypertension, trauma, and long hospital stay are risk factors for MDRO infection in patients with chronic osteomyelitis.

Objective: To explore the impact of tennis on the visual health of primary school students, so as to provide a basis for myopia prevention. Methods: In September 2021, a total of 202 third grade students from Fengtai Affiliated School of Renmin University of China in Beijing were recruited, with 75 students in the intervention group and 127 students in the control group. In addition to attending the same cultural and physical education courses, the intervention group added a tennis course twice a week and tested their vision before the intervention (September 2021), middle (January 2022) and later (June 2022). Data collected at different time points in the two groups were analyzed by using independent sample t-test for comparison between blocks, and single factor repeated measurement ANOVA for analysis of time point factors. Results: After intervention, among male subjects, the average visual acuity of the left and right eyes in the intervention group were higher than that in the control group at the third visual acuity test (left eye:5.00±0.13,4.88±0.29, right eye:5.00±0.12,4.88±0.30, t =2.33, 2.36, P <0.05). Among female subjects, the second visual acuity test (left eye:5.03±0.17, 4.86±0.21, right eye:5.03±0.15, 4.85±0.23) and the third visual acuity test (left eye:4.97±0.13, 4.81±0.23, right eye:4.97±0.14, 4.82±0.24) showed that the average visual acuity of the left and right eyes in the intervention group were higher than that in the control group among females (second: t =3.84, 3.87, third: t =3.70, 3.46, P < 0.01 ). After intervention, the detection rate of visual impairment in both males and females control groups increased at three tests, with statistically significant differences ( χ 2=17.86, 34.77, P <0.01). In the first visual acuity test, there was no statistically significant difference in the detection rate of visual impairment between the control group and the intervention group for both males and females ( χ 2=0.01, 0.10, P >0.05). The third visual impairment detection rate of males in the intervention group was lower than that of the control group (20.00% vs 45.45%), while the second and third visual impairment detection rates of female students in the intervention group were lower than those of the control group (22.86% vs 54.00%; 28.57% vs 70.00%) ( χ 2=7.34, 8.24, 14.18, P < 0.01). Conclusions Tennis can alleviate and prevent the occurrence of myopia in primary school students to a certain extent. Sports such as tennis should be actively promoted to protect the visual health of primary school students.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

Objective:To evaluate the changes of peripapillary retinal and choroid thickness of tail-suspension rats in simulated weightless.Methods:Sixty rats were divided into 2 groups: control group (15 rats) and tail-suspension group (45 rats). The rats in tail-suspension group were tested by -30° tail suspension to simulate the effect of weightlessness for establishing model. The peripapillary retinal thickness of the rats were measured before and after 4- and 8-week tail suspension by enhanced depth imaging-optical coherence tomography (EDI-OCT). The choroid thickness of the rats were measured before and after 4-, 8-, 10-, and 12-week tail-suspension by EDI-OCT.Results:There was a significant difference in the peripapillary retinal thickness among the groups with different suspension time ( F=30.89, P<0.001), and the thickness showed a downward trend with the extension of the tail suspension time. After 8 weeks of tail-suspension, there was a significant difference in the peripapillary retinal thickness between the tail-suspension group and the control group ( t=5.73, P<0.001). After 4, 8, 10, and 12 weeks of tail-suspension, there were significant differences in the choroidal thickness of the rats between the tail-suspension group and the control group ( t=6.32, 12.78, 9.69, 6.39, all P<0.001); there was a significant difference in the choroidal thickness of the rats with different suspension time ( F=51.08, P<0.001). With the time of the tail suspension, the choroidal thickness and its changes compared with before tail suspension showed an upward trend, and reached peak after 8 weeks and then slightly decreased. Conclusions:With the extension of tail-suspension time, the changes of peripapillary retinal thickness of the tail-suspension rats generally show a downward trend, and the changes of choroidal thickness generally increased, and decreased slightly after reaching peak.