ObjectiveTo understand the seropositivity of neutralizing antibodies (NAb) and low-level NAb against SARS-CoV-2 infection in the community residents, and to explore the impact of COVID-19 vaccination and SARS-CoV-2 infection on the levels of NAb in human serum. MethodsOn the ground of surveillance cohort for acute infectious diseases in community populations in Shanghai, a proportional stratified sampling method was used to enroll the subjects at a 20% proportion for each age group (0‒14, 15‒24, 25‒59, and ≥60 years old). Blood samples collection and serum SARS-CoV-2 NAb concentration testing were conducted from March to April 2023. Low-level NAb were defined as below the 25^th percentile of NAb. ResultsA total of 2 230 participants were included, the positive rate of NAb was 97.58%, and the proportion of low-level NAb was 25.02% (558/2 230). Multivariate logistic regression analysis indicated that age, infection history and vaccination status were correlated with low-level NAb (all P<0.05). Individuals aged 60 years and above had the highest risk of low-level NAb. There was a statistically significant interaction between booster vaccination and one single infection (aOR=0.38, 95%CI: 0.19‒0.77). Compared to individuals without vaccination, among individuals infected with SARS-CoV-2 once, both primary immunization (aOR=0.23, 95%CI: 0.16‒0.35) and booster immunization (aOR=0.12, 95%CI: 0.08‒0.17) significantly reduced the risk of low-level NAb; among individuals without infections, only booster immunization (aOR=0.28, 95%CI: 0.14‒0.52) showed a negative correlation with the risk of low-level NAb. ConclusionsThe population aged 60 and above had the highest risk of low-level NAb. Regardless of infection history, a booster immunization could reduce the risk of low-level NAb. It is recommended that eligible individuals , especially the elderly, should get vaccinated in a timely manner to exert the protective role of NAb.

Objective:To assess the clinical value of global RNA N6-methyladenosine (m 6A) and perilipin 2 (PLIN2) mRNA site-specific (chr9:19116312) m 6A modification in peripheral blood as novel molecular biomarkers of coronary artery disease (CAD). Methods:Seventy-four patients with coronary artery disease diagnosed at the Eastern Theater General Hospital from June to December 2023, and 60 age-and sex-matched healthy controls during the same period were selected for a retrospective case-control study. The global RNA m 6A modification level in peripheral blood was detected by RNA methylation quantitative detection kit as a preliminary validation, and PLIN2 site-specific (chr9:19116312) m 6A modification level was further detected using the qPCR quantification technique with single-base elongation and ligation as a rescreening validation. Lipid indicators such as total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C), routine blood indicators such as neutrophil count and platelet count were detected, and the coronary lesion characteristics were further evaluated by using the Gensini score and SYNTAX score systems in conjunction with coronary arteriography results.The CAD group was divided into a single-branch (25 cases) and a multi-branch lesion subgroup (49 cases) according to the number of vascular lesion branches on coronary angiography. The potential value of global RNA m 6A and PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood of patients for the adjunctive diagnosis and assessment of coronary artery disease was explored using Spearman correlation analysis, subject operating characteristic (ROC) curves, and logistic regression analysis. Results:Compared with the control group, the levels of global RNA m 6A modification and PLIN2 site-specific (chr9∶19116312) m 6A modification in peripheral blood were significantly decreased in the CAD group (both P<0.05). The levels of global RNA m 6A modification in peripheral blood was also significantly decreased in the single-branch and multi-branch lesion subgroups ( P<0.05), and PLIN2 site-specific (chr9:19116312) m 6A modification in peripheral blood was significantly decreased in the multi-branch lesion subgroup only ( P<0.05). Spearman correlation analysis showed that in both the CAD group and the multi-branch lesion subgroup, global RNA m 6A modification level in peripheral blood was positively correlated with HDL-C ( r=0.246, 0.289, P<0.05) and negatively correlated with SYNTAX score ( r=-0.261, -0.322, P<0.05) and neutrophil count ( r=-0.246, -0.466, P<0.05). In the single-branch lesion subgroup of CAD, PLIN2 site-specific (chr9:19116312) m 6A modification level was negatively correlated with Gensini score ( r=-0.566, P<0.05). In the multi-branch lesion subgroup of CAD, PLIN2 site-specific (chr9:19116312) m 6A modification level was negatively correlated with platelet count ( r=-0.313, P<0.05). The ROC curve analysis showed that the area under the ROC curve (AUC) of global RNA m 6A and PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood for distinguishing CAD and coronary artery multi-branch lesions were 0.915 and 0.918, with specificity of 83.3% and 95.0%, and sensitivity of 85.1% and 75.5%, respectively. The multivariate logistic regression analysis showed that after adjusting confounding factors such as age, sex, proportion of diabetes and hypertension, and TC, the levels of global RNA m 6A modification ( OR=0.691, P<0.001; OR=0.694, P<0.01), and PLIN2 site-specific (chr9:19116312) m 6A modification levels ( OR=0.345, P<0.05; OR=0.143, P<0.01) in peripheral blood remained independently associated with CAD and coronary artery multi-branch lesions, respectively. Conclusions:The analysis of global RNA m 6A in combination with PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood is valuable for the adjunctive diagnosis and assessment of patients with CAD and coronary artery multi-branch lesions.

ObjectiveTo elucidate the critical role of rehabilitation medical records (including electronic records) in rehabilitation medicine's clinical practice and management, comprehensively analyzed the structure, core content and data standards of rehabilitation medical records, to develop a standardized medical record data architecture and core dataset suitable for rehabilitation medicine and to explore the application of rehabilitation data in performance evaluation and payment. MethodsBased on the regulatory documents Basic Specifications for Medical Record Writing and Basic Specifications for Electronic Medical Records (Trial) issued by National Health Commission of China, and referencing the World Health Organization (WHO) Family of International Classifications (WHO-FICs) classifications, International Classification of Diseases (ICD-10/ICD-11), International Classification of Functioning, Disability and Health (ICF), and International Classification of Health Interventions (ICHI Beta-3), this study constructed the data architecture, core content and data standards for rehabilitation medical records. Furthermore, it explored the application of rehabilitation record summary sheets (home page) data in rehabilitation medical statistics and payment methods, including Diagnosis-related Groups (DRG), Diagnosis-Intervention Packet (DIP) and Case Mix Index. ResultsThis study proposed a systematic standard framework for rehabilitation medical records, covering key components such as patient demographics, rehabilitation diagnosis, functional assessment, rehabilitation treatment prescriptions, progress evaluations and discharge summaries. The research analyzed the systematic application methods and data standards of ICD-10/ICD-11, ICF and ICHI Beta-3 in the fields of medical record terminology, coding and assessment. Constructing a standardized data structure and data standards for rehabilitation medical records can significantly improve the quality of data reporting based on the medical record summary sheet, thereby enhancing the quality control of rehabilitation services, effectively supporting the optimization of rehabilitation medical insurance payment mechanisms, and contributing to the establishment of rehabilitation medical performance evaluation and payment based on DRG and DIP. ConclusionStructured rehabilitation records and data standardization are crucial tools for quality control in rehabilitation. Systematically applying the three reference classifications of the WHO-FICs, and aligning with national medical record and electronic health record specifications, facilitate the development of a standardized rehabilitation record architecture and core dataset. Standardizing rehabilitation care pathways based on the ICF methodology, and developing ICF- and ICD-11-based rehabilitation assessment tools, auxiliary diagnostic and therapeutic systems, and supporting terminology and coding systems, can effectively enhance the quality of rehabilitation records and enable interoperability and sharing of rehabilitation data with other medical data, ultimately improving the quality and safety of rehabilitation services.

Objective To analyze the serum high-density lipoprotein 3 subtype cholesterol(HDL3-C)levels in patients with ischemic stroke(IS)and explore its clinical value in evaluating the condition and prognosis.Methods A total of 124 patients with IS and 47 healthy controls admitted to the Department of Neurology of the General Hospital of the Eastern Theater Command were continuously se-lected from January to June 2023.The serum samples of IS patients at admission and healthy controls were collected respectively,and the levels of HDL3-C,total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipopro-tein cholesterol(LDL-C),blood glucose(Glu),albumin(Alb),interleukin 6(IL-6)and procalcitonin(PCT)were measured.The National Institutes of Health Stroke Scale(NIHSS)score at admission,modified Rankin Scale(MRS)score at discharge,and Barthel index of the IS patients were calculated.All the IS patients were followed up for 1 year.The time of sequelae caused by stroke,and re-currence of acute cerebrovascular events were recorded in detail.Spearman's correlation analysis was used to explore the correlation be-tween HDL3-C levels and other clinical events and biochemical parameters in the IS patients.Logistic regression analysis was used to analyze the clinical value of serum HDL3-C levels in evaluating the presence of IS.Cox regression analysis was used to analyze the clin-ical value of serum HDL3-C levels in assessing the prognosis of patients with IS.Results Compared with controls,the levels of HDL3-C,HDL-C and Alb of the patients with IS decreased significantly,but the levels of TG and Glu of the IS patients increased sig-nificantly(all P＜0.05).The level of serum HDL3-C in patients with IS was significantly positively correlated with TC(r=0.231,P=0.003),HDL-C(r=0.831,P＜0.001),Alb(r=0.451,P＜0.001)and Barthel index at discharge(r=0.216,P=0.018),while it was significantly negatively correlated with the levels of TG(r=-0.396,P＜0.001),IL-6(r=-0.290,P=0.013),NIHSS at admis-sion(r=-0.187,P=0.041)and MRS at discharge(r=-0.227,P=0.012).Multivariate Logistic regression analyses showed that the decreased level of serum HDL3-C was still independently related to the presence of IS(OR=0.853,95％CI=0.740-0.984,P=0.030)after adjusting for the fectors of age,sex,TC,TG,HDL-C and LDL-C.Multivariate Cox regression analyses demonstrates that the de-creased level of serum HDL3-C was still independently related to the poor prognosis of IS patients(HR=0.710,95％CI=0.517-0.976,P=0.035)after adjusting for the fectors of age,sex,TC,TG,HDL-C and LDL-C.Conclusion The level of serum HDL3-C in pa-tients with IS decreased significantly,and it is independently correlated with poor prognosis.The level of HDL3-C has reference value in evaluating the condition and prognosis of the patients with IS.

Objective To analyze the serum high-density lipoprotein 3 subtype cholesterol(HDL3-C)levels in patients with ischemic stroke(IS)and explore its clinical value in evaluating the condition and prognosis.Methods A total of 124 patients with IS and 47 healthy controls admitted to the Department of Neurology of the General Hospital of the Eastern Theater Command were continuously se-lected from January to June 2023.The serum samples of IS patients at admission and healthy controls were collected respectively,and the levels of HDL3-C,total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipopro-tein cholesterol(LDL-C),blood glucose(Glu),albumin(Alb),interleukin 6(IL-6)and procalcitonin(PCT)were measured.The National Institutes of Health Stroke Scale(NIHSS)score at admission,modified Rankin Scale(MRS)score at discharge,and Barthel index of the IS patients were calculated.All the IS patients were followed up for 1 year.The time of sequelae caused by stroke,and re-currence of acute cerebrovascular events were recorded in detail.Spearman's correlation analysis was used to explore the correlation be-tween HDL3-C levels and other clinical events and biochemical parameters in the IS patients.Logistic regression analysis was used to analyze the clinical value of serum HDL3-C levels in evaluating the presence of IS.Cox regression analysis was used to analyze the clin-ical value of serum HDL3-C levels in assessing the prognosis of patients with IS.Results Compared with controls,the levels of HDL3-C,HDL-C and Alb of the patients with IS decreased significantly,but the levels of TG and Glu of the IS patients increased sig-nificantly(all P＜0.05).The level of serum HDL3-C in patients with IS was significantly positively correlated with TC(r=0.231,P=0.003),HDL-C(r=0.831,P＜0.001),Alb(r=0.451,P＜0.001)and Barthel index at discharge(r=0.216,P=0.018),while it was significantly negatively correlated with the levels of TG(r=-0.396,P＜0.001),IL-6(r=-0.290,P=0.013),NIHSS at admis-sion(r=-0.187,P=0.041)and MRS at discharge(r=-0.227,P=0.012).Multivariate Logistic regression analyses showed that the decreased level of serum HDL3-C was still independently related to the presence of IS(OR=0.853,95％CI=0.740-0.984,P=0.030)after adjusting for the fectors of age,sex,TC,TG,HDL-C and LDL-C.Multivariate Cox regression analyses demonstrates that the de-creased level of serum HDL3-C was still independently related to the poor prognosis of IS patients(HR=0.710,95％CI=0.517-0.976,P=0.035)after adjusting for the fectors of age,sex,TC,TG,HDL-C and LDL-C.Conclusion The level of serum HDL3-C in pa-tients with IS decreased significantly,and it is independently correlated with poor prognosis.The level of HDL3-C has reference value in evaluating the condition and prognosis of the patients with IS.

Objective:To assess the clinical value of global RNA N6-methyladenosine (m 6A) and perilipin 2 (PLIN2) mRNA site-specific (chr9:19116312) m 6A modification in peripheral blood as novel molecular biomarkers of coronary artery disease (CAD). Methods:Seventy-four patients with coronary artery disease diagnosed at the Eastern Theater General Hospital from June to December 2023, and 60 age-and sex-matched healthy controls during the same period were selected for a retrospective case-control study. The global RNA m 6A modification level in peripheral blood was detected by RNA methylation quantitative detection kit as a preliminary validation, and PLIN2 site-specific (chr9:19116312) m 6A modification level was further detected using the qPCR quantification technique with single-base elongation and ligation as a rescreening validation. Lipid indicators such as total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C), routine blood indicators such as neutrophil count and platelet count were detected, and the coronary lesion characteristics were further evaluated by using the Gensini score and SYNTAX score systems in conjunction with coronary arteriography results.The CAD group was divided into a single-branch (25 cases) and a multi-branch lesion subgroup (49 cases) according to the number of vascular lesion branches on coronary angiography. The potential value of global RNA m 6A and PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood of patients for the adjunctive diagnosis and assessment of coronary artery disease was explored using Spearman correlation analysis, subject operating characteristic (ROC) curves, and logistic regression analysis. Results:Compared with the control group, the levels of global RNA m 6A modification and PLIN2 site-specific (chr9∶19116312) m 6A modification in peripheral blood were significantly decreased in the CAD group (both P<0.05). The levels of global RNA m 6A modification in peripheral blood was also significantly decreased in the single-branch and multi-branch lesion subgroups ( P<0.05), and PLIN2 site-specific (chr9:19116312) m 6A modification in peripheral blood was significantly decreased in the multi-branch lesion subgroup only ( P<0.05). Spearman correlation analysis showed that in both the CAD group and the multi-branch lesion subgroup, global RNA m 6A modification level in peripheral blood was positively correlated with HDL-C ( r=0.246, 0.289, P<0.05) and negatively correlated with SYNTAX score ( r=-0.261, -0.322, P<0.05) and neutrophil count ( r=-0.246, -0.466, P<0.05). In the single-branch lesion subgroup of CAD, PLIN2 site-specific (chr9:19116312) m 6A modification level was negatively correlated with Gensini score ( r=-0.566, P<0.05). In the multi-branch lesion subgroup of CAD, PLIN2 site-specific (chr9:19116312) m 6A modification level was negatively correlated with platelet count ( r=-0.313, P<0.05). The ROC curve analysis showed that the area under the ROC curve (AUC) of global RNA m 6A and PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood for distinguishing CAD and coronary artery multi-branch lesions were 0.915 and 0.918, with specificity of 83.3% and 95.0%, and sensitivity of 85.1% and 75.5%, respectively. The multivariate logistic regression analysis showed that after adjusting confounding factors such as age, sex, proportion of diabetes and hypertension, and TC, the levels of global RNA m 6A modification ( OR=0.691, P<0.001; OR=0.694, P<0.01), and PLIN2 site-specific (chr9:19116312) m 6A modification levels ( OR=0.345, P<0.05; OR=0.143, P<0.01) in peripheral blood remained independently associated with CAD and coronary artery multi-branch lesions, respectively. Conclusions:The analysis of global RNA m 6A in combination with PLIN2 site-specific (chr9:19116312) m 6A modification levels in peripheral blood is valuable for the adjunctive diagnosis and assessment of patients with CAD and coronary artery multi-branch lesions.

Objective:To investigate the positive rate and isolate Bordetella pertussis in children with suspected whooping cough and their close family members in Zhejiang Province, and further explore the susceptibility and resistant mechanism of Bordetella pertussis to antibiotics. Methods:A total of 273 nasopharynx swabs specimens from children with suspected whooping cough in Hangzhou Children′s Hospital from May 2022 to October 2022 were collected. The strains were isolated and cultured using charcoal select agar plate. Pertussis target genes were detected by RT-PCR. E-test method was used to detect the sensitivity of Bordetella pertussis strains to different antibiotics. The mechanism of resistance of Bordetella pertussis to macrolides was analyzed by whole genome sequencing. The phylogenetic analysis of isolated strains was based on core genome multilocus sequence typing(cgMLST). Results:Among 273 clinical samples of children with suspected pertussis and their close family members, 168 samples were positive by fluorescence quantitative PCR, accounting for 61.54%, and 30 pertussis strains were successfully isolated with a positive rate of 10.98%. In addition, among the 143 samples of close family members, 54.55% (78/143) samples were positive by RT-PCR and 9.79% (14/143) samples were positive by culture, suggesting that the close family member are important in family transmission of pertussis. Besides, most of the positive samples were from mothers. The results of E-test showed that 96.67%(29/30) strains showed high resistance to azithromycin with MIC value>256 mg/L, and the resistant mechanism of azithromycin was A2047G mutation in 23S rRNA. The phylogenetic analysis based on the cgMSLT showed that the isolated strains were clustered into two new different clades.Conclusions:The positive rate of Bordetella pertussis in close family members is at a high level and the mother may be the main source of infection, which is of great significance for monitoring and laboratory detection of suspected children′s family members. Bordetella pertussis shows high resistance to macrolides in Zhejiang Province, so monitoring of the antimicrobial resistance should be further strengthened to provide theoretical basis for clinical treatment and drug guidance.

Objective:To identify the phosphodiesterase (PDE) activity of the gene product encoded by LA_RS06960 of Leptospira interrogans ( L. interrogans), and analyze whether dinucleotides that can be degraded by PDE can activate macrophages to express innate immune factors. Methods:The LA_RS06960 gene in L. interrogans strain 56601 was amplified by PCR, and the prokaryotic expression system was constructed for the protein expression. The expressed rPDE was purified by Ni-NTA affinity chromatography. High performance liquid chromatography (HPLC) was used to measure the degration of c-di-AMP or 5′-pApA to AMP by rPDE. Real-time fluorescent quantitative RT-PCR (qRT-PCR) was used to detect the changes in the expression of target genes in Leptospira or THP-1 cells associated with innate immune factors during infection. qRT-PCR and ELISA were used to detect the changes in the expression and secretion level of the innate immune factors in macrophages treated with bacterial dinucleotide. Results:The prokaryotic expression system for LA_RS06960 gene of L. interrogans was constracted successfully, and the purified rPDE could degrate 5′-pApA and c-di-AMP into AMP in vitro. The mRNA level of leptospiral LA_RS06960 gene was significantly down-regulated, while IFN-β, TNF-α, IL-6 and IL-1β encoding genes of macrophages were significantly up-regulated during infection. The mRNA level or the secretion level of IFN-β and IL-6 of macrophages were increased after treated with the bacterial dinucleotide substrate of PDE. Conclusions:PDE encoded by LA_RS06960 gene has phosphodiesterase activity, and the bacterial dinucleotide substrate of the PDE could activate the innate immune response of macrophages.

Objective:To detect the changes in the levels of miR-21-5p and miR-125a-5p modified with 3′-terminal 2′-O-methylation (3′t-2′Ome) in serum extracellular vesicles (EV) of non-small cell lung cancer (NSCLC) patients, and evaluate their value as auxiliary screening molecular markers for NSCLC patients.Method:A retrospective analysis was conducted on the data of 69 NSCLC patients diagnosed at the Eastern Theater Command General Hospital from May 1st to October 31st,2023, as well as 65 age and gender matched healthy controls during the same period. Two real-time fluorescence quantitative PCR (RT-qPCR) techniques, namely stem-loop method and poly (A) tailed method, were used to detect the levels of 3′t-2′Ome-miR-21-5p and 3′t-2′-Ome miR-125a-5p in serum EV of NSCLC patients and controls.Analyze the correlation between the levels of two types of 3′t-2 ′Ome miRNAs and the differences in clinical stage, pathological classification, and other tumor indicators in patients. Receiver operating characteristic (receiveroperating curve, ROC) curves were used to analyze the efficacy of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combination, in diagnosing NSCLC.Result:Compared with the control group, the levels of 3′t-2′Ome-miR-21-5p in serum EV of NSCLC patients increased [(0.30±0.05) vs (0.35±0.09), t=3.32, P=0.001], while the levels of 3′t-2′Ome-miR-125a-5p decreased [(0.33±0.06 vs 0.25±0.06, t=7.45, P<0.001]. The differences were statistically significant. The expression levels of 2′Ome-miR-21-5p in EV were also significantly elevated in the NSCLC patients at 0-Ⅱ stage, adenocarcinoma patients, and squamous cell carcinoma patients, respectively. Notably, the levels of EV 3′t-2′Ome-miR-21-5p was also statistically significant between the adenocarcinoma patients and squamous cell carcinoma patients [(0.34±0.85) vs (0.40±0.12), P<0.05]. ROC curve analysis showed that the levels of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combined AUC for discriminating NSCLC patients, were 0.647(95% CI 0.550-0.743), 0.825(95% CI 0.756-0.894) and 0.860(95% CI 0.797-0.923), respectively. The sensitivity was 92.3%, 80.0%, 89.2%, and the specificity was 46.4%, 73.9%, and 78.3%, respectively. Conclusion:There are changes in the levels of 2 ′Ome miR-21-5p and 2′ Ome miR-125a-5p in the serum EV of NSCLC patients, and the combined detaction has the potential as an auxiliary screening molecular marker of NSCLC patients.

ObjectiveTo present the exploration and application of a prospective follow-up research method for acute infectious disease surveillance based on natural community populations， using COVID-19 infection as an example， and to provide a reference for improving the infectious disease surveillance and early warning system. MethodsA multi-stage probability proportional sampling method was employed to sample residents from all communities of 16 administrative districts in Shanghai， with households as the units. A cohort for acute infectious diseases based on natural community populations was established. The baseline survey was conducted for all cohort subjects， and COVID-19 antigen test kits were distributed. From December 21， 2022 to September 30， 2023， prospective follow-up monitoring of COVID-19 antigen and nucleic acid was carried out on the study subjects on a weekly basis. The baseline characteristics and follow-up information of the cohort subjects were described. ResultsThe cohort for acute infectious diseases included a total of 12 881 subjects， comprising 6 098 males （47.3%） and 6 783 females （52.7%）. The baseline survey revealed that 35.2% （4 540/12 881） of the subjects had a history of COVID-19 infection. During the follow-up period from December 21， 2022 to September 30， 2023， the average incidence density in the cohort was 0.61/person-year， with a higher incidence density in females （0.63/person-year） compared to males （0.59/person-year）. Individuals aged 60 and above （0.64/person-year） and those with underlying health conditions （0.67/person-year） had a higher incidence density. Healthcare workers showed a notably higher incidence density （0.84/person-year） than that in other occupational groups. As of September 30， 2023， a total of 340 subjects in the cohort experienced secondary infections， with a median interval of 170 days between the first and second infections. ConclusionThis study applies cohort study method to acute infectious disease surveillance， providing crucial data support for estimating infection rates and forecasting alerts for acute infectious diseases in the community. This method can be promoted and applied as a new approach for acute infectious disease surveillance.