This study aimed to explore the interaction between the small molecule Lyb24 and PyrD, a key enzyme in the pyrimidine biosynthesis pathway of Klebsiella pneumoniae (KP), and the effect of Lyb24 on the catalytic activity of PyrD, thus to provide a theoretical basis for the development of novel antimicrobial agents. The pET-30a(+)-PyrD recombinant plasmid was constructed using Nde I/Xba I double digestion technology and was transformed into Escherichia coli BL21 (DE3) competent cells using the heat-shock method. The recombinant protein was induced at 16 ℃ with 0.3 mmol/L isopropyl β-D-thiogalactopyranoside (IPTG). The recombinant PyrD protein was purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography to obtain a high-purity product. Surface plasmon resonance (SPR) experiments were conducted to detect the direct interaction between Lyb24 and PyrD protein, and a DCIP-based colorimetric assay was used to evaluate the effect of Lyb24 on the catalytic activity of PyrD. The pET-30a(+)-PyrD plasmid was successfully constructed, and the recombinant PyrD protein with a molecular weight of approximately 36 kD was expressed and purified to a concentration of 5.58 mg/mL. Lyb24 exhibited high-affinity direct binding to PyrD (KD = 8.83 × 10−5 mol/L) and exerted an uncompetitive inhibition effect on the catalytic activity of PyrD. This study demonstrates that Lyb24, a small-molecule compound, directly binds to PyrD and inhibits its enzymatic activity, providing crucial experimental evidence for developing PyrD-targeted antibacterial agents with value of clinical translation.

Objective: To explore the relationship between visual acuity and physical fitness of university freshmen, so as to provide reference for myopia prevention and control for freshmen. Methods: From October to November 2022, 2 160 college freshman without majoring in public safety administration, selected from Guangxi Police College in 2022 by using the stratified cluster random sampling method, were reviewed for the results of visual acuity test and physical fitness scores. The physical fitness indices were evaluated by using the Z scores of physical fitness test scores, and the strength of association between the level of physical fitness index and myopia was analyzed by using Logistic regression model. Results: Among 2 160 college freshman without majoring in public safety administration, 917 (42.5%) students were diagnosed screening myopia, including 66 (3.1%) cases of high myopia, 383 (17.7%) cases of moderate myopia and 468 (21.7%) cases of mild myopia. The differences in the distribution of visual acuity tests among students with different physical fitness indices, body mass index, and gender were statistically significant ( Z/H=54.50, 49.53, 15.51, P <0.01). Low level and low middle level physical fitness indices were associated with screening myopia among freshmen[ OR (95% CI )=2.81(1.93-4.08),1.87(1.38-2.54)], and low level physical fitness indexes were associated with high myopia [ OR (95% CI )=7.22(2.33-22.32)] ( P <0.01). Conclusions Screening myopia among college freshman without majoring in public safety administration is related to physical fitness, and low level and low middle level physical fitness index are risk factors for myopia. Improving the level of physical fitness might be effective in preventing myopia.

Objective:To investigate the effect of TCF1+CD8+T cells on the prognosis of nasopharyngeal carcinoma(NPC)pa-tients undergoing immunotherapy.Methods:A retrospective study was conducted on 108 NPC patients admitted to Anyang Cancer Hospital from January 2018 to April 2020.Pathological findings and clinical data were collected,and multiple immunofluorescence staining was utilized to detect TCF1+CD8+T cell levels in tumor tissue.Cell counts and the proportion of positive cells were measured,and the optimal cut-off value was determined using a time-dependent receiver operating characteristic(ROC)curve.Patients were clas-sified into high expression group and low expression group according to the cut-off value.The general information and survival status were compared between two groups,and univariate and multivariate Cox regression analysis were performed to screen the influencing factors of patient prognosis.Results:The optimal cut-off value was determined using ROC curve.A cut-off value of 0.34%for TCF1+CD8+T cell area under the curve(AUC)=0.653 resulted in a sensitivity of 73.5%and a specificity of 43.7%.Based on the opti-mal cut-off value,33 patients with TCF1+CD8+T cell beyond 0.34%were included in high expression group,and the remaining 75 pa-tients were included in low expression group.The proportion of TCF1+CD8+T cells was(0.43±0.09)%in high expression group and(0.21±0.08)%in low expression group.Comparison of general data yielded that the gender,age,body mass index(BMI),clinical stage,and pathological classification showed no statistical difference between two groups(P>0.05),while the percentage of patients with T stage of T3 and T4,N stage of N2 and N3,and M stage of M1 in high expression group were smaller than that in low expression group(P<0.05).The 24-month survival rate was 81.82%(27/33)in high expression group and 74.67%(56/75)in low expression group,with no statistical difference between two groups(χ2=0.658 8,P=0.417 0).Univariate and multivariate Cox regression analysis found that T stage,N stage and M stage were independent risk factors affecting the prognosis of patients,and TCF1+CD8+T expression was a protective factor.ROC curve indicated that the AUC,sensitivity and specificity of TCF1+CD8+T expression in predicting death of NPC patients undergoing immunotherapy was 0.674,69.7%and 65.3%,while the AUC,sensitivity and specificity of TCF1+CD8+T ex-pression combined with TNM stage in predicting death was 0.809,75.8%and 84.0%,respectively.Conclusion:NPC patients with higher proportion of TCF1+CD8+T cells and lower TNM stage have higher 24-month survival rate after immunotherapy,and the above parameters are of great predictive value for patient prognosis.

Objective:To investigate the survival outcomes and prognostic factors of patients with double primary breast cancer (BC) and endometrial cancer (EC).Methods:A retrospective cohort study was conducted using data for the period 1992-2018 from the Surveillance, Epidemiology, and End Results (SEER) database. There were 3 465 patients with BC as the first primary cancer (BC-EC group) and 2 804 patients with EC as the first primary cancer (EC-BC group). Kaplan-Meier analysis and cumulative incidence function were used to estimate overall mortality, breast cancer-specific mortality, and endometrial cancer-specific mortality, respectively. Cox regression and Fine-Gray regression were used to analyze the prognostic factors of overall mortality, breast cancer-specific mortality, and endometrial cancer-specific mortality, respectively.Results:During a median follow-up of 160 months, 1 616 deaths occurred in the BC-EC group, with EC being the leading cause of death (37.69%); 994 deaths occurred in the EC-BC group, with BC being the leading cause of death (28.77%). Cox regression identified patients with older ages at first primary cancer diagnosis (54-61 years: HR=1.46, 95% CI: 1.26-1.69; 62-68 years: HR=2.64, 95% CI: 2.29-3.03; ≥69 years: HR=4.89, 95% CI: 4.27-5.60), shorter time interval between the diagnoses (0-5 months: HR=6.13, 95% CI: 5.21-7.21; 6-23 months: HR=5.69, 95% CI: 4.95-6.55; 24-59 months: HR=3.44, 95% CI: 3.04-3.89; 60-119 months: HR=2.32, 95% CI: 2.07-2.59), mixed ductal-lobular BC ( HR=1.29, 95% CI: 1.11-1.48), endometrial mixed cell adenocarcinoma ( HR=1.23, 95% CI: 1.01-1.50), advanced tumor grade (grade Ⅱ BC: HR=1.13, 95% CI: 1.01-1.27; grade Ⅲ BC: HR=1.24, 95% CI: 1.10-1.41; grade Ⅱ EC: HR=1.19, 95% CI: 1.06-1.33; grade Ⅲ EC: HR=1.68, 95% CI: 1.48-1.90), advanced tumor stage of the two cancers (distant BC: HR=3.14, 95% CI: 2.50-3.94; regional EC: HR=1.53, 95% CI: 1.36-1.71; distant EC: HR=3.00, 95% CI: 2.59-3.47) had increased risk of overall mortality. Fine-Gray regression showed that compared with BC-EC patients, EC-BC patients had a higher risk of breast cancer-specific mortality [sub-distribution hazard ratio (s HR=1.24, 95% CI: 1.04-1.47], but a lower risk of endometrial cancer-specific mortality (s HR=0.37, 95% CI: 0.30-0.46). Older ages at first cancer diagnosis, shorter intervals between the diagnoses, negative ER and PR status, and advanced BC grades/stages were associated with increased breast cancer-specific mortality ( P＜0.05). Similarly, older ages, shorter intervals, endometrial serous carcinoma/mixed cell adenocarcinoma, and advanced EC grades/stages correlated with elevated endometrial cancer-specific mortality ( P＜0.05). Conclusion:The management of double primary BC and EC patients requires multidisciplinary strategies, with particular attention to patients presenting older ages at first cancer diagnosis, shorter intervals between the diagnoses, and unfavorable tumor characteristics.

Objective:To investigate the effect of TCF1+CD8+T cells on the prognosis of nasopharyngeal carcinoma(NPC)pa-tients undergoing immunotherapy.Methods:A retrospective study was conducted on 108 NPC patients admitted to Anyang Cancer Hospital from January 2018 to April 2020.Pathological findings and clinical data were collected,and multiple immunofluorescence staining was utilized to detect TCF1+CD8+T cell levels in tumor tissue.Cell counts and the proportion of positive cells were measured,and the optimal cut-off value was determined using a time-dependent receiver operating characteristic(ROC)curve.Patients were clas-sified into high expression group and low expression group according to the cut-off value.The general information and survival status were compared between two groups,and univariate and multivariate Cox regression analysis were performed to screen the influencing factors of patient prognosis.Results:The optimal cut-off value was determined using ROC curve.A cut-off value of 0.34%for TCF1+CD8+T cell area under the curve(AUC)=0.653 resulted in a sensitivity of 73.5%and a specificity of 43.7%.Based on the opti-mal cut-off value,33 patients with TCF1+CD8+T cell beyond 0.34%were included in high expression group,and the remaining 75 pa-tients were included in low expression group.The proportion of TCF1+CD8+T cells was(0.43±0.09)%in high expression group and(0.21±0.08)%in low expression group.Comparison of general data yielded that the gender,age,body mass index(BMI),clinical stage,and pathological classification showed no statistical difference between two groups(P>0.05),while the percentage of patients with T stage of T3 and T4,N stage of N2 and N3,and M stage of M1 in high expression group were smaller than that in low expression group(P<0.05).The 24-month survival rate was 81.82%(27/33)in high expression group and 74.67%(56/75)in low expression group,with no statistical difference between two groups(χ2=0.658 8,P=0.417 0).Univariate and multivariate Cox regression analysis found that T stage,N stage and M stage were independent risk factors affecting the prognosis of patients,and TCF1+CD8+T expression was a protective factor.ROC curve indicated that the AUC,sensitivity and specificity of TCF1+CD8+T expression in predicting death of NPC patients undergoing immunotherapy was 0.674,69.7%and 65.3%,while the AUC,sensitivity and specificity of TCF1+CD8+T ex-pression combined with TNM stage in predicting death was 0.809,75.8%and 84.0%,respectively.Conclusion:NPC patients with higher proportion of TCF1+CD8+T cells and lower TNM stage have higher 24-month survival rate after immunotherapy,and the above parameters are of great predictive value for patient prognosis.

Objective:To investigate the survival outcomes and prognostic factors of patients with double primary breast cancer (BC) and endometrial cancer (EC).Methods:A retrospective cohort study was conducted using data for the period 1992-2018 from the Surveillance, Epidemiology, and End Results (SEER) database. There were 3 465 patients with BC as the first primary cancer (BC-EC group) and 2 804 patients with EC as the first primary cancer (EC-BC group). Kaplan-Meier analysis and cumulative incidence function were used to estimate overall mortality, breast cancer-specific mortality, and endometrial cancer-specific mortality, respectively. Cox regression and Fine-Gray regression were used to analyze the prognostic factors of overall mortality, breast cancer-specific mortality, and endometrial cancer-specific mortality, respectively.Results:During a median follow-up of 160 months, 1 616 deaths occurred in the BC-EC group, with EC being the leading cause of death (37.69%); 994 deaths occurred in the EC-BC group, with BC being the leading cause of death (28.77%). Cox regression identified patients with older ages at first primary cancer diagnosis (54-61 years: HR=1.46, 95% CI: 1.26-1.69; 62-68 years: HR=2.64, 95% CI: 2.29-3.03; ≥69 years: HR=4.89, 95% CI: 4.27-5.60), shorter time interval between the diagnoses (0-5 months: HR=6.13, 95% CI: 5.21-7.21; 6-23 months: HR=5.69, 95% CI: 4.95-6.55; 24-59 months: HR=3.44, 95% CI: 3.04-3.89; 60-119 months: HR=2.32, 95% CI: 2.07-2.59), mixed ductal-lobular BC ( HR=1.29, 95% CI: 1.11-1.48), endometrial mixed cell adenocarcinoma ( HR=1.23, 95% CI: 1.01-1.50), advanced tumor grade (grade Ⅱ BC: HR=1.13, 95% CI: 1.01-1.27; grade Ⅲ BC: HR=1.24, 95% CI: 1.10-1.41; grade Ⅱ EC: HR=1.19, 95% CI: 1.06-1.33; grade Ⅲ EC: HR=1.68, 95% CI: 1.48-1.90), advanced tumor stage of the two cancers (distant BC: HR=3.14, 95% CI: 2.50-3.94; regional EC: HR=1.53, 95% CI: 1.36-1.71; distant EC: HR=3.00, 95% CI: 2.59-3.47) had increased risk of overall mortality. Fine-Gray regression showed that compared with BC-EC patients, EC-BC patients had a higher risk of breast cancer-specific mortality [sub-distribution hazard ratio (s HR=1.24, 95% CI: 1.04-1.47], but a lower risk of endometrial cancer-specific mortality (s HR=0.37, 95% CI: 0.30-0.46). Older ages at first cancer diagnosis, shorter intervals between the diagnoses, negative ER and PR status, and advanced BC grades/stages were associated with increased breast cancer-specific mortality ( P＜0.05). Similarly, older ages, shorter intervals, endometrial serous carcinoma/mixed cell adenocarcinoma, and advanced EC grades/stages correlated with elevated endometrial cancer-specific mortality ( P＜0.05). Conclusion:The management of double primary BC and EC patients requires multidisciplinary strategies, with particular attention to patients presenting older ages at first cancer diagnosis, shorter intervals between the diagnoses, and unfavorable tumor characteristics.

Background Bacteria are the most diverse and widely sourced microorganisms in the indoor air of subway stations, where pathogenic bacteria can spread through the air, leading to increased health risks. Objective To understand the status and distribution characteristics of indoor air bacterial pollution in subway stations and compartments in a city of Central South China, and to provide a scientific basis for formulating intervention measures to address indoor air bacteria pollution in subways. Methods Three subway stations and the compartments of trains parking there in a city in Central South China were selected according to passenger flow for synchronous air sampling and monitoring. Temperature, humidity, wind speed, carbon dioxide (CO₂), fine particulate matter (PM_2.5), and inhalable particulate matter (PM₁₀) were measured by direct reading method. In accordance with the requirements of Examination methods for public places-Part 3: Airborne microorganisms (GB/T 18204.3-2013), air samples were collected at a flow rate of 28.3 L·min⁻¹, and total bacterial count was estimated. Bacterial microbial species were identified with a mass spectrometer and pathogenic bacteria were distinguished from non-pathogenic bacteria according to the Catalogue of pathogenic microorganisms transmitted to human beings issued by National Health Commission. Kruskal-Wallis H test was used to compare the subway hygiene indicators in different regions and time periods, and Bonferroni test was used for pairwise comparison. Spearman correlation test was used to evaluate the correlation between CO₂ concentration and total bacterial count. Results The pass rates were 100.0% for airborne total bacteria count, PM_2.5, and PM₁₀ in the subway stations and train compartments, 94.4% for temperature and wind speed, 98.6% for CO₂, but 0% for humidity. The overall median (P₂₅, P₇₅) total bacteria count was 177 (138,262) CFU·m⁻³. Specifically, the total bacteria count was higher in station halls than in platforms, and higher during morning peak hours than during evening peak hours (P<0.05). A total of 874 strains and 82 species were identified by automatic microbial mass spectrometry. The results of identification were all over 9 points, and the predominant bacteria in the air were Micrococcus luteus (52.2%) and Staphylococcus hominis (9.8%). Three pathogens, Acinetobacter baumannii (0.3%), Corynebacterium striatum (0.1%), and Staphylococcus epidermidis bacilli (2.2%) were detected in 23 samples (2.6%), and the associated locations were mainly distributed in train compartments during evening rush hours. Conclusion The total bacteria count in indoor air varies by monitoring sites of subway stations and time periods, and there is a risk of opportunistic bacterial infection. Attention should be paid to cleaning and disinfection during peak passenger flow hours in all areas.

Objective:To explore the rehabilitation effect of multi-factor intervention based on the Finnish model of prevention of cognitive impairment in the elderly on patients with cognitive impairment after first-episode stroke, and to provide reference for rehabilitation nursing of cognitive impairment after stroke.Methods:The quasi-experiment research scheme was adopted and convenience sampling method was used to select participants with first-episode stroke cognitive impairment hospitalized in the General Hospital of Tianjin Medical University Airport Site. The 50 patients admitted from January to June 2022 were selected as the control group, and 50 patients admitted from July to December 2022 were selected as the intervention group. The control group received routine rehabilitation nursing and health education, and the intervention group received the Finnish model of prevention of cognitive impairment in the elderly on patients before discharge on the basis of the control group. The Mini-Mental State Examination (MMSE) and Health Education Compliance Assessment Scale for Stroke Patients were used to evaluate the changes of overall cognitive function and rehabilitation compliance before intervention, 3 and 6 months after intervention.Results:The final control group included 49 cases, including 35 males and 14 females, aged (64.67 ± 7.47) years old; the intervention group included 50 cases, 32 males and 18 females, aged (66.68 ± 8.75) years old. Before intervention, there were no significant differences in overall cognitive function and compliance of rehabilitation score ( P>0.05). At 3 and 6 months after intervention, the overall cognitive function score, the total score on compliance of rehabilitation, dimension scores of diet compliance, exercise rehabilitation compliance and health behavior compliance of the intervention group were (26.36±2.36) , (125.96 ± 13.80) , (23.30 ± 5.26) , (27.72 ± 4.46) , (43.66 ± 6.80) and (27.26 ± 3.71) , (152.44 ± 9.06) , (30.12 ± 6.42) , (33.32 ± 3.02) , (52.36 ± 4.70) , respectively. They were higher than the control group (24.04 ± 4.50) , (116.67 ± 10.26) , (19.31 ± 3.95) , (25.29 ± 3.45) , (40.59 ± 4.33) and (24.27 ± 4.33) , (138.92 ± 16.71) , (24.20 ± 4.48) , (30.00 ± 5.53) , (47.65 ± 8.03) , and the differences had statistical significance ( t values were -5.31- -2.67, all P<0.05). According to the variance analysis of repeated measurement, intergroup and time factor, the interaction between groups and time had significant impact on general cognitive function score, the total score of rehabilitation compliance, the dimension scores of diet, exercise rehabilitation and health behavior compliance ( Fgroup values were 8.33-18.08, Ftime values were 135.71-944.69, Finteraction values were 5.46-27.30, all P<0.05) . Time factor had significant impact on patient medication adherence score ( Ftime=206.23, P<0.05) . Conclusions:Multi-factor intervention based on the Finnish model of prevention of cognitive impairment in the elderly can improve the overall cognitive function and rehabilitation compliance of patients with cognitive impairment after first-episode stroke.

Lipid Droplets/metabolism* ; Bacteria ; Lipid Metabolism

Objective:To investigate the impact of human epidermal growth factor receptor 2 (HER2) and C-reactive protein/albumin ratio (CAR) on the long-term survival of gastric cancer patients after surgery.Methods:A retrospective analysis was conducted on 136 gastric cancer patients admitted to Lishui Central Hospital from January 2015 to December 2017, who underwent radical surgery and were followed up for 5 years. Patients were divided into HER2 positive and HER2 negative groups based on HER2 immunohistochemical results, and into high CAR and low CAR groups based on the CAR mean value. The relationship between HER2 and CAR with clinicopathological characteristics of gastric cancer patients was analyzed. The postoperative tumor-free survival rate and overall survival rate were compared between the two groups of patients (HER2 positive group and HER2 negative group, as well as high CAR group and low CAR group). Cox regression analysis was used to identify independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients.Results:The proportion of HER2 positive patients with large tumor size, low differentiation, T 3-4 tumor invasion depth, lymph node metastasis, and vascular invasion was significantly higher than that of HER2 negative patients (all P<0.05). The proportion of high CAR patients with large tumor size, low differentiation, T 3-4 tumor invasion depth, lymph node metastasis, and vascular invasion was significantly higher than that of low CAR patients (all P<0.05). Kaplan-Meier survival analysis showed that HER2 negative patients had significantly higher 1-year, 3-year, and 5-year cumulative tumor-free survival rate and overall survival rate than HER2 positive patients, while low CAR patients had significantly higher 1-year, 3-year, and 5-year cumulative tumor-free survival rate and overall survival rate than high CAR patients (all P<0.05). Multivariate Cox regression analysis identified T 3-4 tumor invasion depth, lymph node metastasis, HER2 positivity, and high CAR expression as independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients (all P<0.05). HER2 positive gastric cancer patients had a 1.895-fold higher risk of postoperative tumor recurrence and metastasis than HER2 negative patients ( HR: 1.895, 95% CI: 1.245-4.229, P=0.034), while high CAR gastric cancer patients had a 1.769-fold higher risk of postoperative tumor recurrence and metastasis than low CAR patients ( HR: 1.769, 95% CI: 1.433-3.959, P=0.039). HER2 positive gastric cancer patients had a 2.145-fold higher risk of postoperative death than HER2 negative patients ( HR: 2.145, 95% CI: 1.378-4.589, P=0.028), while high CAR gastric cancer patients had a 1.926-fold higher risk of postoperative death than low CAR patients ( HR: 1.926, 95% CI: 1.564-3.853, P=0.025). Conclusions:HER2 and CAR are independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients. Gastric cancer patients with HER2 positivity and high CAR have a higher risk of postoperative tumor recurrence, metastasis, and death. This study has some limitations due to its small sample size and single-center design, which may introduce some bias. Future multicenter and large-scale studies are needed to confirm the results of this study.