BACKGROUND:The pathogenesis of osteoporosis is closely related to the immune system.A comprehensive and in-depth study of the relationship between immunity and osteoporosis is crucial for understanding and treating the disease.OBJECTIVE:To investigate the role of immune cells in osteoporosis using single-cell sequencing technology.METHODS:Femoral head tissue samples from osteoporosis and non-osteoporosis patients were downloaded from GEO database and analyzed using single-cell sequencing.Data analysis,including cell clustering,functional enrichment,pseudotime trajectory,and cell interaction analyses,was performed using R4.3.0 and software packages such as Seurat v.4.3,monocle(2.28.0),and CellChat.The femoral head tissues of patients with femoral neck fracture who underwent artificial hip replacement surgery were obtained,including two cases of osteoporosis patients and two cases of non-osteoporosis patients.Immunohistochemical staining was used to detect the protein expression of CCL13 and CCL18.qPCR was used to detect the immunoglobulin heavy constant γ-4,immunoglobulin λ constant 3,human class Ⅱ major histocompatibility complex DRβ1,and CD83 mRNA expression.Western blot was used to detect the protein expression of receptor-type tyrosine protein phosphatase C,CD22,and CD99.RESULTS AND CONCLUSION:Transcriptomic analysis identified 10 cell clusters,including osteoclasts,myeloid cells,T cells,osteoblasts,macrophages,monocytes,erythrocytes,B cells,bone marrow mesenchymal stem cells,and mast cells.There was an increase in the ratio of osteoclasts to T cells and a decrease in the ratio of osteoblasts to B cells in the femoral head tissue of the osteoporosis group.Among the B-cell subpopulations,the proportion of B-cells of taxa 1,3(BC1,BC3)in the femoral head tissue of the osteoporosis group was higher than that of the non-osteoporosis group,and the proportion of B-cells of taxa 2(BC2)was less than that of the non-osteoporosis group.BC1 was enriched significantly for labels such as regulation of adaptive immune response,somatic recombination of immune receptors,and modulation of lymphocyte-mediated immunity,while BC3 was enriched significantly for labels such as regulation of immunoglobulin production,response to type Ⅱ interferon,apoptotic processes involving cysteine endopeptidases,and cytotoxicity.The communication intensity between B-cell subtype BC1 and osteoblasts in the femoral head tissue of the osteoporosis group was higher than that of the non-osteoporosis group,while the communication intensity between BC3 and BC1 was also increased.The communication between BC3 and BC1 was significantly enriched in the CD22-receptor-type tyrosine protein phosphatase C pathway;the communication between BC1 and osteoblasts was mainly enriched in the CD99-CD99 pathway;and the communication between BC3 and osteoblasts was also highly enriched in the CD99-CD99 pathway.Protein expression of CCL13,CCL18,receptor-type tyrosine protein phosphatase C,CD22,CD99,immunoglobulin heavy constant γ-4,immunoglobulin λ constant 3,human class Ⅱ major histocompatibility complex DRβ1,and CD83 mRNA were higher in femoral tissues of the osteoporosis group than those of the non-osteoporosis group(P＜0.05).To conclude,specific B cell subpopulations can influence the differentiation and apoptosis of osteoblasts in the femoral tissue of osteoporosis patients.

Objective:To investigate the association between frailty and prognosis of elderly patients in the emergency department, and to validate frailty screening tools suitable for the emergency department.Methods:This was a prospective cohort study. Clinical data of elderly patients over 60 years old treated in the emergency department of Beijing Bo'Ai Hospital from January to December 2021 were collected. The Frailty Screening Questionnaire (FSQ), FRAIL Scale (FRAIL) and Clinical Frailty Scale (CFS) were used to score patients, and patients were divided into frail or non-frail group according to the criteria of the above three scales. Twelve-month all-cause mortality was the primary endpoint, dependence and re-admission to the emergency department within 12 months were secondary outcomes. Receiver operating characteristic curves were used to evaluate the ability of the FSQ, FRAIL and CFS scores to predict the primary and secondary endpoints, and the areas under the curve (AUC) were calculated and compared. Survival analysis was performed using Cox hazard proportional regression model, and relative risk was expressed as hazard ratio ( HR) and 95% CI. Results:A total of 406 patients were included in the study. The AUCs (95% CI) of FSQ, FRAIL and CFS scores for predicting 12-month all-cause mortality were 0.879 (0.844-0.909), 0.838 (0.798-0.872), 0.906 (0.873-0.933), respectively (all P<0.001). The AUCs of 3 scores for predicting secondary endpoints ranged from 0.820 to 0.889 (all P<0.001). Pairwise comparisons of the AUCs showed that the CFS was superior to one or both of the other frailty screening scales in predicting 12-month all-cause mortality and dependence except for re-admission to emergency room within 12 months after discharge (all P<0.05). Cox regression analysis revealed that, after adjusting for sex, age, body mass index and comorbidities, frailty as defined by the FSQ, FRAIL, and CFS scales was independently associated with 12-month all-cause mortality, with the HRadj of 3.267 (95% CI: 2.406-4.435), 2.465 (95% CI: 1.819-3.341), 3.523 (95% CI: 2.648-4.687), respectively (all P<0.001). Conclusions:FSQ, FRAIL and CFS scores can predict adverse outcomes, the CFS is a practical frailty screening tool in the emergency department, and frailty screening can improve the risk stratification of older patients.

Objective:To explore the association between acute muscle wasting during hospitalization and poor prognosis in older patients with severe community-acquired pneumonia (SCAP) in emergency department.Methods:This study was a prospective cohort study. From January 1, 2022 to October 31, 2022, consecutive patients aged ≥65 years who met the diagnostic criteria of SCAP and had an interval of 14 days between two CT scans in the emergency department of Beijing Chao-Yang Hospital were enrolled. The general clinical data and cross-sectional area of the erector spinae muscle (ESMcsa) of the thoracic 12 level derived from chest CT on day 1 and day 14 were recorded and the differences between the two measurements were calculated. Patients were divided into survival group and non-survival group based on whether they died within 28 days. Two independent samples t-test and Mann Whitney U test were used to compare the dynamic changes of ESMcsa between two groups, and paired t-test and Wilcoxon signed rank test were used to compare the changes of ESMcsa within two groups. Multivariable Cox regression analysis was used to identify the risk factors for 28-day mortality, and receiver operating characteristic (ROC) curves were used to determine the predictive value of ESMcsa loss for 28-day mortality. The optimal cutoff value was determined on the basis of the Youden index (YI), patients were divided into a high muscle loss group and a low muscle loss group, and Kaplan Meier survival curve was drawn. Results:A total of 106 older patients with SCAP were included, with a median age of 82.0 years and 59 were men (55.7%). The ESMcsa levels of patients in non-survival group were lower than those in survival group both at admission and on the 14th day (both P<0.01). The ESMcsa levels on admission were lower than those on the 14th day in non-survival group ( P<0.001). The loss of ESMcsa in non-survival group [3.01 (-1.51, 7.73) cm 2vs. 0.80 (-2.58, 4.57) cm 2, P=0.020] was higher than that in the survival group. Multivariable Cox regression showed that ESMcsa loss was an independent risk factor for 28-day mortality ( HR=1.116, 95%CI: .029-1.210, P=0.010), the AUC for predicting 28-day mortality was 0.646 (95% CI: 0.528-0.763, P=0.020), and the optimal cut-off value was 6.22 cm 2. Kaplan Meier survival curve showed that the 28-day mortality risk in the high muscle loss group was higher than that in the low muscle loss group ( χ2=11.412, P=0.001). Conclusion:Acute muscle wasting during hospitalization was associated with 28-day mortality among older patients with SCAP, which provides a basis for improving patient prognosis from a muscle perspective.

Objective:To develop and validate a prediction model by combining clinical data and biomarkers to evaluate the probability of frailty among older emergency patients.Methods:A cross-sectional study was conducted. From January 2021 to December 2021, patients aged 60 years and older admitted to the emergency department of China Rehabilitation Research Center were enrolled. Data of patient's clinical information were collected. The patients were divided into frail group and non-frail group according to the Fried's frailty phenotype and clinical data were compared between the two groups. LASSO regression was used to deal with dimension reduction and multivariate logistic regression was employed to construct a prediction model based on variables selected by the LASSO regression. Nomogram was used to visualize the prediction model. The area under the receiver operating characteristic curve, calibration curve, decision curve analysis and bootstrap were used to evaluate the discrimination, calibration, clinical applicability, and internal validity of the model respectively.Results:A total of 348 patients were enrolled, and the incidence of frailty was 53.74% (187/348). Education, coronary heart disease, chronic obstructive pulmonary disease, albumin, fibrinogen, N-terminal pro-brain natriuretic peptide, decreased creatinine, and underweight were independent predictors for frailty in older emergency patients ( P < 0.05). A nomogram model was built based on the above predictors and the model showed good discrimination, calibration and clinical applicability. Conclusions:The study utilized objective clinical data and biomarkers to establish a predictive model for the occurrence of frailty in elderly emergency department patients. This model aids in risk stratification and targeted intervention for elderly emergency patients, thereby improving patient outcomes.

Objective:To understand the species of mosquitoes and the status of important mosquito-borne viruses in the Ningxia surveyed regions, to identify the dominant mosquito species and virus types, and to analyze their genetic characteristics, providing a scientific basis for predicting and controlling mosquito-borne infectious diseases.Methods:Mosquitoes were collected using light traps in Yinchuan and Wuzhong cities of the Ningxia Hui Autonomous Region, and the collected mosquitoes were classified and identified. Real-time polymerase chain reaction (PCR) was used to detect the Japanese encephalitis virus (JEV), West Nile virus (WNV), Chikungunya virus (CHIKV), Sindbis virus (SINV) carried by mosquitoes. The positive sample was subjected to sequencing the whole genome, and the phylogenetic tree of virus strains was constructed using bioinformatics methods.Results:From June to August 2023, a total of 8 561 mosquitoes of 3 genera and 6 species were collected in Yinchuan and Wuzhong cities, Ningxia, among which Culex pipiens pallens was the dominant species with 3 050 individuals, accounting for 35.63%; Anopheles sinensis with 2 379 individuals, accounting for 27.79%; Culex tritaeniorhynchus with 1 489 individuals, accounting for 17.39%; Caspian Aedes with 1 468 individuals, accounting for 27.79%; Aedes vexans with 152 individuals, accounting for 1.78%; and Culex modestus with 23 individuals, accounting for 0.27%. JEV GIb type was detected in the specimens of Culex tritaeniorhynchus collected in Qingtongxia city. Conclusions:The dominant mosquito species in the surveyed areas of Ningxia are primarily Culex pipiens pallens, and JEV GIb virus was detected in Culex tritaeniorhynchus in Qingtongxia city. This study provides basic data for understanding the current status of mosquitoes and mosquito-borne viruses in the Ningxia region and offers scientific guidance for further public health prevention and control measures.

Objective:To understand the species of mosquitoes and the status of important mosquito-borne viruses in the Ningxia surveyed regions, to identify the dominant mosquito species and virus types, and to analyze their genetic characteristics, providing a scientific basis for predicting and controlling mosquito-borne infectious diseases.Methods:Mosquitoes were collected using light traps in Yinchuan and Wuzhong cities of the Ningxia Hui Autonomous Region, and the collected mosquitoes were classified and identified. Real-time polymerase chain reaction (PCR) was used to detect the Japanese encephalitis virus (JEV), West Nile virus (WNV), Chikungunya virus (CHIKV), Sindbis virus (SINV) carried by mosquitoes. The positive sample was subjected to sequencing the whole genome, and the phylogenetic tree of virus strains was constructed using bioinformatics methods.Results:From June to August 2023, a total of 8 561 mosquitoes of 3 genera and 6 species were collected in Yinchuan and Wuzhong cities, Ningxia, among which Culex pipiens pallens was the dominant species with 3 050 individuals, accounting for 35.63%; Anopheles sinensis with 2 379 individuals, accounting for 27.79%; Culex tritaeniorhynchus with 1 489 individuals, accounting for 17.39%; Caspian Aedes with 1 468 individuals, accounting for 27.79%; Aedes vexans with 152 individuals, accounting for 1.78%; and Culex modestus with 23 individuals, accounting for 0.27%. JEV GIb type was detected in the specimens of Culex tritaeniorhynchus collected in Qingtongxia city. Conclusions:The dominant mosquito species in the surveyed areas of Ningxia are primarily Culex pipiens pallens, and JEV GIb virus was detected in Culex tritaeniorhynchus in Qingtongxia city. This study provides basic data for understanding the current status of mosquitoes and mosquito-borne viruses in the Ningxia region and offers scientific guidance for further public health prevention and control measures.

BACKGROUND:The pathogenesis of osteoporosis is closely related to the immune system.A comprehensive and in-depth study of the relationship between immunity and osteoporosis is crucial for understanding and treating the disease.OBJECTIVE:To investigate the role of immune cells in osteoporosis using single-cell sequencing technology.METHODS:Femoral head tissue samples from osteoporosis and non-osteoporosis patients were downloaded from GEO database and analyzed using single-cell sequencing.Data analysis,including cell clustering,functional enrichment,pseudotime trajectory,and cell interaction analyses,was performed using R4.3.0 and software packages such as Seurat v.4.3,monocle(2.28.0),and CellChat.The femoral head tissues of patients with femoral neck fracture who underwent artificial hip replacement surgery were obtained,including two cases of osteoporosis patients and two cases of non-osteoporosis patients.Immunohistochemical staining was used to detect the protein expression of CCL13 and CCL18.qPCR was used to detect the immunoglobulin heavy constant γ-4,immunoglobulin λ constant 3,human class Ⅱ major histocompatibility complex DRβ1,and CD83 mRNA expression.Western blot was used to detect the protein expression of receptor-type tyrosine protein phosphatase C,CD22,and CD99.RESULTS AND CONCLUSION:Transcriptomic analysis identified 10 cell clusters,including osteoclasts,myeloid cells,T cells,osteoblasts,macrophages,monocytes,erythrocytes,B cells,bone marrow mesenchymal stem cells,and mast cells.There was an increase in the ratio of osteoclasts to T cells and a decrease in the ratio of osteoblasts to B cells in the femoral head tissue of the osteoporosis group.Among the B-cell subpopulations,the proportion of B-cells of taxa 1,3(BC1,BC3)in the femoral head tissue of the osteoporosis group was higher than that of the non-osteoporosis group,and the proportion of B-cells of taxa 2(BC2)was less than that of the non-osteoporosis group.BC1 was enriched significantly for labels such as regulation of adaptive immune response,somatic recombination of immune receptors,and modulation of lymphocyte-mediated immunity,while BC3 was enriched significantly for labels such as regulation of immunoglobulin production,response to type Ⅱ interferon,apoptotic processes involving cysteine endopeptidases,and cytotoxicity.The communication intensity between B-cell subtype BC1 and osteoblasts in the femoral head tissue of the osteoporosis group was higher than that of the non-osteoporosis group,while the communication intensity between BC3 and BC1 was also increased.The communication between BC3 and BC1 was significantly enriched in the CD22-receptor-type tyrosine protein phosphatase C pathway;the communication between BC1 and osteoblasts was mainly enriched in the CD99-CD99 pathway;and the communication between BC3 and osteoblasts was also highly enriched in the CD99-CD99 pathway.Protein expression of CCL13,CCL18,receptor-type tyrosine protein phosphatase C,CD22,CD99,immunoglobulin heavy constant γ-4,immunoglobulin λ constant 3,human class Ⅱ major histocompatibility complex DRβ1,and CD83 mRNA were higher in femoral tissues of the osteoporosis group than those of the non-osteoporosis group(P＜0.05).To conclude,specific B cell subpopulations can influence the differentiation and apoptosis of osteoblasts in the femoral tissue of osteoporosis patients.

Objectives:To investigate the impacts of American College of Cardiology/American Heart Association (ACC/AHA) coronary artery classification on the progression of coronary non-target lesions and revascularization in patients with coronary heart disease.Methods:From January 2010 to September 2014,1255 patients who underwent two consecutive coronary angiographies at Fuwai Hospital and had coronary non-target lesions were retrospectively analyzed.Lesion characteristics of all coronary non-target lesions were recorded at both procedures.All non-target lesions were divided into A,B1,B2 and C lesion group according to ACC/AHA coronary artery classification.Patients were divided into non-B2/C lesion group (noncomplex lesion group) and B2/C lesion group (complex lesion group) according to whether the non-target lesion had B2/C lesion The characteristics of all non-target coronary artery lesions and quantitative coronary angiography results were recorded.Lesion progression and revascularization were compared between different groups.Results:There were 1003 (79.9％) male patients,mean age was (58.0±9.7) years old,and 853 patients had B2/C lesions.There were 1670 non-target lesions,including 619 A/B1 lesions (214 A lesions and 405 B1 lesions) and 1051 B2/C lesions (796 B2 lesions and 255 C lesions).Follow-up time was (14.8±4.5) months.Compared with the patients in noncomplex lesion group,patients in complex lesion group were older,had lower proportion of family history of coronary heart disease and stroke (all P<0.05).The baseline levels of leukocytes,C-reactive protein,erythrocyte sedimentation rate (ESR),triglyceride and HbA1c were higher in complex lesion group than those in noncomplex lesion group.Complex lesion group had higher risk of lesion progression (21.8％ vs.13.2％,P<0.001) compared with noncomplex lesion group,similar results were observed in revascularization (16.5％ vs.11.2％,P=0.013),and there was no statistically difference in non-target lesion related myocardial infarction (P>0.05).At the lesion level,compared with A/B1 lesion,B2/C lesion was associated with a higher rate of lesion progression (17.4％ vs.11.0％,P<0.001),and a higher rate of revascularization (13.0％ vs.9.2％,P=0.018).Multivariate Cox regression analysis showed that lesion classification (B2/C) was an independent risk factor for non-target lesion progression (HR=1.732,95％CI:1.275-2.351,P<0.001) and non-target lesion revascularization (HR=1.477,95％CI:1.053-2.070,P=0.024).Conclusions:The risk of non-target lesion progression and revascularization is higher in complex groups compared with noncomplex groups according to ACC/AHA classification.So patients with complex lesions should receive more strict medical care to control related risk factors and improve their outcome.

Objective:To establish a 14-day prognosis model for emergency patients with acute ischemic cerebral stroke and evaluate its predictive efficacy.Methods:A prospective cohort study was conducted. Patients with acute ischemic stroke admitted to the emergency department of Beijing Bo’ai Hospital within 72 hours of onset from October 2018 to December 2020 were enrolled. Univariate and multivariate logistic regression analysis were used to screen the risk factors of poor prognosis. The ROC curve was drawn to determine the cut-off value of continuous variables and discretise data with reference to clinical practice. The corresponding scores were set up according to the β regression coefficient of each variable, and the clinical scale prediction model of short-term prognosis of acute cerebral infarction was established. Patients with ischemic stroke in the hospital from January to December 2021 were selected as the internal validation, to verify the constructed predictive model.Results:A total of 321 patients were included in the study, including 223 in the training set and 98 in the internal validation set. Multivariate logistic regression analysis showed that age, hypersensitive C-reactive protein, prealbumin (PA), infarct volume, Frailty Screening Questionnaire (FSQ) and National Institute of Health Stroke Scale (NIHSS) were independent risk factors for poor short-term prognosis of acute cerebral infarction. The total score of the clinical prediction scoring system for short-term prognosis of acute cerebral infarction in the emergency department was 15 points, including age ≥74 years (1 point), PA ≤373 mg/L (2 points), large artery atherosclerosis (1 point), cardiogenic embolism (2 points), infarct volume ≥ 2.18 cm 3 (2 points), FSQ ≥3 points (1 point), NIHSS ≥4 points (6 points); The area under the ROC curve (AUC) of the scoring system for predicting short-term poor prognosis of acute cerebral infarction was 0.927 (95% CI: 0.894-0.960). The optimal cut-off value was ≥5 points, and the sensitivity and specificity were 0.770 and 0.976, respectively. In the internal validation set, the scoring system had similar predictive value for poor outcomes (AUC=0.892, 95% CI:0.827-0.957). Conclusion:The scoring system for short-term prognosis prediction of acute ischemic cerebral infarction has good diagnostic efficacy, and could guide clinicians to judge the prognosis of emergency patients in the early stage.

【Objective】 To investigate the molecular mechanism of microRNA-101-5p (miR-101-5p) affecting the proliferation and invasion of lung squamous cell carcinoma (LUSC) cells. 【Methods】 We downloaded the miRNA mature expression data and total RNA sequencing data of TCGA-LUSC from TCGA database to identify differentially expressed genes. The signal pathway enriched in ATAD2 was analyzed. The mRNA expressions of miR-101-5p and ATAD2 in the LUSC cells were detected by qRT-PCR. The effects of miR-101-5p on the proliferation and invasion of LUSC cells were detected by MTT assay, cloning assay, and invasion assay. The effects of ATAD2 on the cell cycle of LUSC cells were detected by flow cytometry. Western blotting was used to detect the expression of ATAD2 protein. Double luciferase experiment was used to verify whether miR-101-5p could bind to ATAD2 target. Finally, we detected the changes in the proliferation, cloning and invasion ability of LUSC cells by co-transfection with oe-ATAD2 and miR-101-5p mimic, and further explored whether miR-101-5p could regulate the biological function of LUSC cells through ATAD2. 【Results】 The miR-101-5p was significantly downregulated in LUSC tissues and cells. Overexpression of miR-101-5p could significantly inhibit the proliferation and invasion of LUSC cells. ATAD2, its downstream regulatory target gene, was significantly upregulated in LUSC, and miR-101-5p and ATAD2 expressions were inversely correlated. GSEA enrichment results showed that ATAD2 was significantly enriched in the cell cycle signal pathway. The double luciferase experiment proved that miR-101-5p targeted ATAD2, and the recovery experiment showed that miR-101-5p regulated the proliferation and invasion of LUSC cells by targeting ATAD2. 【Conclusion】 In this study, we found that miR-101-5p had low expression in LUSC, and that miR-101-5p decreased the proliferation and invasion of LUSC cells by targeted inhibition of ATAD2.